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Support:

877-678-TECH (8324)

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For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Mol. Wt Isotype/Source
LRP1 (E2Q7S) Rabbit mAb2638720 µl85 kDaRabbit IgG
ApoE (pan) (D7I9N) Rabbit mAb1336620 µl35 kDaRabbit IgG
PSD95 (D27E11) XP® Rabbit mAb345020 µl95 kDaRabbit IgG
Phospho-PSD95 (Ser295) (A8F8Z) Rabbit mAb4573720 µl95 kDaRabbit IgG
Synapsin-1 (D12G5) XP® Rabbit mAb529720 µl77 kDaRabbit IgG
AMPA Receptor 1 (GluA1) (D4N9V) Rabbit mAb1318520 µl100 kDaRabbit IgG
Phospho-AMPA Receptor 1 (GluA1) (Ser831) (A5O2P) Rabbit mAb7557420 µl100 kDaRabbit IgG
CREB (48H2) Rabbit mAb919720 µl43 kDaRabbit IgG
Phospho-CREB (Ser133) (87G3) Rabbit mAb919820 µl43 kDaRabbit IgG
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit provides an economical means of detecting key synaptic signaling pathways in response to ApoE-mediated LRP1 activation by western blot. The kit includes enough antibodies to perform at least two western blot experiments with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibodies.

Background

Low density lipoprotein receptor related protein 1 (LRP1) is a type I transmembrane receptor that mediates the endocytosis of various ligands, including apolipoproteins and tau. Both proteins are genetically and pathologically linked to Alzheimer’s disease (AD) (1,2). Human apolipoprotein E (ApoE) is a component of circulating lipoproteins when three human genetic ApoE variants, ApoE2, ApoE3, and ApoE4, exhibit distinct receptor-binding properties and differentially contribute to AD progression through a cellular mechanism that is poorly understood (2). Altered synaptic signaling is one proposed mechanism that contributes to altered neuronal function, which correlates with disease (3-5). Postsynaptic Density protein 95 (PSD95) is a member of the membrane-associated guanylate kinase (MAGUK) family of proteins that functions as a scaffolding protein to promote assembly and function of the postsynaptic density complex (6,7). At the presynapse, synapsins function to regulate neurotransmitter release (8,9). Dynamic phosphorylation of PSD95 at Ser295 reflects synaptic signaling that may alter synaptic function (10). In addition to PSD95, postsynaptic glutamate receptors, including AMPA-(α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) receptors, can be directly phosphorylated. Phosphorylation of AMPA Receptor 1 (GluA1) at either Ser831 or Ser845 alters AMPA receptor ion channel function to change synaptic efficacy (11). CREB is a bZIP transcription factor that activates target genes through cAMP response elements. CREB is activated by phosphorylation at Ser133 by various signaling pathways including Erk, Ca2+,stress signaling, as well as synaptic signaling (3). 

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

Cell Signaling Technology Logo
Western blot analysis of extracts from mouse brain, rat brain, and rat prefrontal cortex tissues using AMPA Receptor 1 (GluA1) (D4N9V) Rabbit mAb.
undefined Image 1: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Western blot analysis of cell extracts from Hep G2 cells treated with Brefeldin A #9972 (10 ng/ml, 90 min) and human cerebellum using ApoE (pan) (D7I9N) Rabbit mAb.
undefined Image 2: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human kidney using ApoE (pan) (D7I9N) Rabbit mAb.
undefined Image 3: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines and tissues using LRP1 (E2Q7S) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
undefined Image 4: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Western blot analysis of extracts from human cerebellum and rat brain using PSD95 (D27E11) XP® Rabbit mAb.
undefined Image 5: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Western blot analysis of extracts from mouse brain, untreated (-) or λ phosphatase-treated (+), using Phospho-PSD95 (Ser295) (A8F8Z) Rabbit mAb (upper) or PSD95 (D74D3) XP® Rabbit mAb #3409 (lower).
undefined Image 6: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

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Western blot analysis of extracts from CAD cells and neonatal mouse brain using Synapsin-1 (D12G5) XP® Rabbit mAb.
undefined Image 7: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
undefined Image 8: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Simple WesternTM analysis of lysates (0.1 mg/mL) from Mouse Brain cells using Phospho-AMPA Receptor 1 (GluA1) (Ser831) (A5O2P) Rabbit mAb #75574. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
undefined Image 9: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

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Western Blot analysis of extracts from SK-N-MC, COS, NIH/3T3, C6 and Drosophila S2 cells, using CREB (48H2) Rabbit mAb.
undefined Image 10: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
CUT&Tag was performed with 293 cells treated with Forskolin #3828 (30 μM) for 1h and either CREB (48H2) Rabbit mAb or Phospho-CREB (Ser133) (87G3) Rabbit mAb #9198, using CUT&Tag Assay Kit #77552. DNA libraries were prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across NR4A3, a known target gene of both CREB and Phospho-CREB (see additional figure containing ChIP-qPCR data).
undefined Image 11: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Western blot analysis of extracts from SK-N-MC cells, untreated or forskolin- and FGF-treated, using Phospho-CREB (Ser133) (87G3) Rabbit mAb (upper) or CREB (48H2) Rabbit mAb #9197 (lower).
undefined Image 12: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

Cell Signaling Technology Logo
CUT&Tag was performed with 293 cells treated with Forskolin #3828 (30 μM) for 1h and either Phospho-CREB (Ser133) (87G3) Rabbit mAb or CREB (48H2) Rabbit mAb #9197, using CUT&Tag Assay Kit #77552. DNA libraries were prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across NR4A3, a known target gene of both Phospho-CREB and CREB (see additional figure containing ChIP-qPCR data).
undefined Image 13: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit

Simple WesternTM analysis of lysates (0.1 mg/mL) from Mouse Brain cells using AMPA Receptor 1 (GluA1) (D4N9V) Rabbit mAb #13185. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

undefined Image 14: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Western blot analysis of cell extracts from 293T cells, treated with Brefeldin A #9972 (10 ng/ml, 90 min; +) and mock transfected (-) or transfected with a construct expressing full-length human ApoE2 (hApoE2; +), ApoE3 (hApoE3; +), or ApoE4 (hApoE4; +), using ApoE (pan) (D7I9N) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
undefined Image 15: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

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Immunohistochemical analysis of paraffin-embedded liver (left) or brain (right) from C57BL/6NTac (wt/wt, model #B6-F, top) and APOE4 knock-in (hu/hu, model #1549-F, bottom) mice using ApoE (pan) (D7I9N) Rabbit mAb. Mice from Taconic Biosciences, Inc.
undefined Image 16: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Confocal immunofluorescent analysis of MEF-1 cells (left, LRP1 (+/+), positive) or PEA 13 cells (right, LRP1 (-/-), negative) using LRP1 (E2Q7S) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
undefined Image 17: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Confocal immunofluorescent analysis of rat cerebellum and retina using PSD95 (D27E11) XP® Rabbit mAb (red) and Neurofilament-L (DA2) Mouse mAb #2835 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 18: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

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Immunohistochemical analysis of paraffin-embedded human cerebellum using Synapsin-1 (D12G5) XP® Rabbit mAb.
undefined Image 19: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Western blot analysis of extracts from mouse brain, untreated (-) or λ-phosphatase-treated (+), using Phospho-AMPA Receptor 1 (GluA1) (Ser831) (A5O2P) Rabbit mAb (upper) and AMPA Receptor 1 (GluA1) (D4N9V) Rabbit mAb #13185 (lower).
undefined Image 20: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Immunoprecipitation of CREB from SK-N-MC extracts. Lane 1 is CREB (48H2) Rabbit mAb, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is 10% input. Western blot analysis was perfomed using CREB (86B10) Mouse mAb #9104. Anti-mouse IgG, HRP-linked Antibody #7076 was used as a secondary antibody.
undefined Image 21: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

Cell Signaling Technology Logo
CUT&Tag was performed with 293 cells treated with Forskolin #3828 (30 μM) for 1h and either CREB (48H2) Rabbit mAb or Phospho-CREB (Ser133) (87G3) Rabbit mAb #9198, using CUT&Tag Assay Kit #77552. DNA libraries were prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 9 (upper), including NR4A3 (lower), a known target gene of both CREB and Phospho-CREB (see additional figure containing ChIP-qPCR data).
undefined Image 22: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, showing nuclear staining, using Phospho-CREB (Ser133) (87G3) Rabbit mAb.
undefined Image 23: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
CUT&Tag was performed with 293 cells treated with Forskolin #3828 (30 μM) for 1h and either Phospho-CREB (Ser133) (87G3) Rabbit mAb or CREB (48H2) Rabbit mAb #9197, using CUT&Tag Assay Kit #77552. DNA libraries were prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 9 (upper), including NR4A3 (lower), a known target gene of both Phospho-CREB and CREB (see additional figure containing ChIP-qPCR data).
undefined Image 24: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

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Immunoprecipitation of AMPA Receptor 1 (GluA1) from mouse brain extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or AMPA Receptor 1 (GluA1) (D4N9V) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using AMPA Receptor 1 (GluA1) (D4N9V) Rabbit mAb.
undefined Image 25: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Immunoprecipitation of ApoE from Hep G2 cells treated with Brefeldin A #9972 (10 ng/ml, 90 min), using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or ApoE (pan) (D7I9N) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using ApoE (pan) (D7I9N) Rabbit mAb.
undefined Image 26: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human spleen using ApoE (pan) (D7I9N) Rabbit mAb.
undefined Image 27: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

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Immunohistochemical analysis of paraffin-embedded mouse cortex using Synapsin-1 (D12G5) XP® Rabbit mAb.
undefined Image 28: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human astrocytoma, using CREB (48H2) Rabbit mAb.
undefined Image 29: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded mouse lung using Phospho-CREB (Ser133) (87G3) Rabbit mAb.
undefined Image 30: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

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Confocal immunofluorescent analysis of mouse hippocampus using AMPA Receptor 1 (GluA1) (D4N9V) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 31: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human small intestine using ApoE (pan) (D7I9N) Rabbit mAb.
undefined Image 32: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded rat retina using Synapsin-1 (D12G5) XP® Rabbit mAb.
undefined Image 33: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

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Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using CREB (48H2) Rabbit mAb.
undefined Image 34: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-CREB (Ser133) (87G3) Rabbit mAb in the presence of control peptide (left) or Phospho-CREB (Ser133) Blocking Peptide #1090 (right).
undefined Image 35: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human skin using ApoE (pan) (D7I9N) Rabbit mAb in the presence of control peptide (left) and antigen-specific peptide (right).
undefined Image 36: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

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Confocal immunofluorescent analysis of mouse brain using Synapsin-1 (D12G5) XP® Rabbit mAb (green) and β3-Tubulin (TU-20) Mouse mAb #4466 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 37: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin's lymphoma, using CREB (48H2) Rabbit mAb.
undefined Image 38: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded SK-N-MC cells, untreated (left) or IBMX- and forskolin-treated (right), showing induced nuclear staining, using Phospho-CREB (Ser133) (87G3) Rabbit mAb.
undefined Image 39: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

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Immunohistochemical analysis of paraffin-embedded 293T cell pellets, control (left-top) or transfected with human ApoE isoforms ApoE2 (right-top), ApoE3 (left-bottom), and ApoE4 (right-bottom) using ApoE (pan) (D7I9N) Rabbit mAb.
undefined Image 40: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded mouse brain, using CREB (48H2) Rabbit mAb.
undefined Image 41: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma, untreated (left) or lambda phosphatase-treated (right), using Phospho-CREB (Ser133) (87G3) Rabbit mAb.
undefined Image 42: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

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Immunohistochemical analysis of paraffin-embedded Hep G2 cell pellet (left, positive) or 293T cell pellet (right, negative) using ApoE (pan) (D7I9N) Rabbit mAb.
undefined Image 43: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Confocal immunofluorescent analysis of mouse cerebellum labeled with CREB (48H2) Rabbit mAb (red) and Neurofilament-L (DA2) Mouse mAb #2835 (green). Blue pseudocolor =DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 44: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Confocal immunofluorescent images of rat dentate gyrus, either sham-operated (left) or 15 min ischemia followed by 30 min (center) and 4 h (right) reperfusion, labeled with Phospho-CREB (Ser133) (87G3) Rabbit mAb (red), Neurofilament-L (DA2) Mouse mAb #2835 (blue) and Phospho-S6 Ribosomal Protein (Ser235/236) (2F9) Rabbit mAb (Alexa Fluor® 488 Conjugate) #4854.
undefined Image 45: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

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Confocal immunofluorescent analysis of Hep G2 cells (left, positive) and 293T cells (right, negative) using ApoE (pan) (D7I9N) Rabbit mAb #13366 (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
undefined Image 46: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Confocal immunofluorescent analysis of SK-N-MC cells showing nuclear stain with CREB (48H2) Rabbit mAb (A, red) compared to an isotype control (B). Blue pseudocolor =DRAQ5® (fluorescent DNA dye).
undefined Image 47: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Confocal microscopic images of SK-N-MC cells showing nuclear stain after 25 minute treatment with Forskolin and IBMX using Phospho-CREB (Ser133) (87G3) Rabbit mAb (left, red) compared to untreated cells (right). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 48: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

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Flow cytometric analysis of Jurkat cells using CREB (48H2) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
undefined Image 49: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with TPA #4174 (500nM) and Ionomycin #9995 (1uM,4hrs;green) using Phospho-CREB (Ser133) (87G3) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
undefined Image 50: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 μM) for 1h and either Phospho-CREB (Ser133) (87G3) Rabbit mAb or CREB (48H2) Rabbit mAb (#9197), using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across NR4A3, a known target gene of both Phospho-CREB and CREB (see additional figure containing ChIP-qPCR data).
undefined Image 51: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

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Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells, treated with Forskolin #3828 (30 μM) for 1h and either 10 μl of CREB (48H2) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human ALS2 exon 1 primers, SimpleChIP® Human NR4A3 Promoter Primers #4829, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
undefined Image 52: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 μM) for 1h and either Phospho-CREB (Ser133) (87G3) Rabbit mAb or CREB (48H2) Rabbit mAb (#9197), using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 9 (upper), including NR4A3 (lower), a known target gene of both Phospho-CREB and CREB (see additional figure containing ChIP-qPCR data).
undefined Image 53: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 μM) for 1h and either Phospho-CREB (Ser133) (87G3) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human ALS2 exon 1 primers, SimpleChIP® Human NR4A3 Promoter Primers #4829, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
undefined Image 54: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

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Western blot analysis of extracts from HepG2 cells (lane 1), 293T mock transfected (lane 2) or transiently transfected with a construct expressing ApoE4 (lane 3), whole liver extracts from wild type C57BL/6NTac model #B6-F mice (lane 4), or ApoE4 knock-in (model #1549-F) (lane 5), whole brain extracts from wild type C57BL/6NTac mice (lane 6), or ApoE4 knock-in (lane 7) using ApoE (pan) (D7I9N) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Mice from Taconic Biosciences, Inc.
undefined Image 55: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
CUT&RUN was performed with 293 cells treated with Forskolin #3828 (30 μM) for 1h and Phospho-CREB (Ser133) (87G3) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across NR4A3 gene.
undefined Image 56: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
CUT&RUN was performed with 293 cells treated with Forskolin #3828 (30 μM) for 1h and Phospho-CREB (Ser133) (87G3) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® (ChIP-Seq, CUT&RUN) #56795. The figures show binding across chromosome 9 (upper), including NR4A3 (lower) gene.
undefined Image 57: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

Cell Signaling Technology Logo
CUT&RUN was performed with 293 cells treated with Forskolin #3828 (30 μM) for 1h and either Phospho-CREB (Ser133) (87G3) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human ALS2 exon 1 primers and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
undefined Image 58: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Confocal immunofluorescent analysis of fixed frozen hippocampus from C57BL/6 (left) and APOE4 knock-in (hu/hu, model #1549-F, right) mice labeled with ApoE (pan) (D7I9N) Rabbit mAb (top, green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to colabeling with GFAP (GA5) Mouse mAb (Alexa Fluor® 594 Conjugate) #8152 (bottom, red), and AQP4 (D1F8E) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #89851 (bottom, blue pseudocolor). Mice from Taconic Biosciences, Inc.
undefined Image 59: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Simple Western™ analysis of lysates (0.1 mg/mL) from mouse brain using PSD95 (D27E11) XP® Rabbit mAb #3450. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66-440 kDa separation module.
undefined Image 60: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of human iPSC-derived cortical glutamatergic neurons at 7 days in vitro using PSD95 (D27E11) XP® Rabbit mAb (green), β3-Tubulin (E9F3E) Mouse mAb #45058 (red), and DAPI #4083 (blue). iCell GlutaNeurons were kindly provided by FUJIFILM Cellular Dynamics, Inc.
undefined Image 61: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Simple WesternTM analysis of lysates (0.75 mg/mL) from Mouse Brain cells using Phospho-PSD95 (Ser295) (A8F8Z) Rabbit mAb #45737. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66-440 kDa separation module.
undefined Image 62: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Simple Western™ analysis of lysates (0.1 mg/mL) from CAD cells using Synapsin-1 (D12G5) XP Rabbit mAb #5297. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
undefined Image 63: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

Cell Signaling Technology Logo
CUT&RUN was performed with 293 cells treated with Forskolin #3828 (30 μM) for 1h and either CREB (48H2) Rabbit mAb or Phospho-CREB (Ser133) (87G3) Rabbit mAb #9198, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across NR4A3 gene.
undefined Image 64: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
CUT&RUN was performed with 293 cells treated with Forskolin #3828 (30 μM) for 1h and either CREB (48H2) Rabbit mAb or Phospho-CREB (Ser133) (87G3) Rabbit mAb #9198, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 9 (upper), including NR4A3 (lower) gene.
undefined Image 65: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
CUT&RUN was performed with 293 cells treated with Forskolin #3828 (30 μM) for 1h and either CREB (48H2) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human ALS2 exon 1 primers and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
undefined Image 66: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

Cell Signaling Technology Logo
Simple Western™ analysis of lysates (0.1 mg/mL) from SK-N-MC cells treated with Forskolin (30uM, 30min) and IBMX (0.5mM, 30 min) using Phospho-CREB (Ser133) (87G3) Rabbit mAb #9198. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. This antibody also detects the phosphorylated form of the CREB-related protein, ATF-1 (as indicated). The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ ​​​​​​​Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
undefined Image 67: ApoE Synaptic Formation and Signaling Pathway Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.