Revision 1

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877-616-CELL (2355)

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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
CBP (D6C5) Rabbit Monoclonal Antibody738920 µl300 kDaRabbit IgG
Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody477120 µl300 kDaRabbit
PCAF (C14G9) Rabbit Monoclonal Antibody337820 µl93 kDaRabbit IgG
GCN5L2 (C26A10) Rabbit Monoclonal Antibody330520 µl94 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 Lysine Acetyltransferase Antibody Sampler Kit provides an economical means to examine several lysine acetyltrasferases, including: Acetyl-CBP, CBP, GCN5L2, and PCAF. The kit contains enough primary antibody to perform two western blots per primary.

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 antibody.

Background

CREB-binding protein (CBP) and p300 are highly conserved and functionally related transcriptional co-activators that associate with transcriptional regulators and signaling molecules, integrating multiple signal transduction pathways with the transcriptional machinery (1,2). CBP/p300 also contain histone acetyltransferase (HAT) activity, allowing them to acetylate histones and other proteins (2). The role of acetylation of CBP/p300 is of particular interest (2,3). Acetylation of p300 at Lys1499 has been demonstrated to enhance its HAT activity and affect a wide variety of signaling events (4). p300/CBP-associated factor (PCAF), also known as lysine acetyl-transferase 2B (KAT2B) (5), and General Control of Amino Acid Synthesis Yeast Homolog Like 2 (GCN5L2) (6) are transcriptional adaptor proteins in addition to HATs. PCAF functions as the catalytic subunit of the PCAF transcriptional co-activator complex (5). GCN5L2 functions as the catalytic subunit of the STAGA and TFTC transcription coactivator complexes (6). PCAF and GCN5L2 acetylate histone H3 at Lys14 and histone H4 at Lys8, both of which contribute to gene activation by modulating chromatin structure and recruiting additional co-activator proteins that contain acetyl-lysine binding bromo-domains (7,8). PCAF also acetylates non-histone proteins including transcriptional activators (p53, E2F1, MyoD) and general transcription factors (TFIIEβ and TFIIF) (9-12). GCN5L2 also acetylates non-histone proteins such as transcription activators (TAT, c-Myb) (13,14), transcription co-activators (PGC1-α) (15), and nuclear receptors (Steroidogenic Factor 1) (16). Acetylation of these proteins regulates their nuclear localization, protein stability, DNA binding, and co-activator association (13-16).

Trademarks and Patents

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

U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using GCN5L2 (C26A10) Rabbit mAb.
undefined Image 1: Lysine Acetyltransferase Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using PCAF (C14G9) Rabbit mAb.
undefined Image 2: Lysine Acetyltransferase Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 µM) for 1 hour and PCAF (C14G9) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across NR4A3, a known target gene of PCAF (see additional figure containing ChIP-qPCR data).
undefined Image 3: Lysine Acetyltransferase 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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from A431, NIH/3T3, COS and PC12 cells, using Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody.
undefined Image 4: Lysine Acetyltransferase 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 5: Lysine Acetyltransferase Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 μM, 1h) and CBP (D6C5) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across NR4A3, a known target gene of CBP (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
undefined Image 6: Lysine Acetyltransferase 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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from control HEK293T cells (Lane 1) or HEK293T cells with a targeted mutation in the gene encoding CBP (Lane 2) using CBP (D6C5) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The change in CBP molecular weight in the mutated HEK293T cells confirms the specificity of the antibody for CBP.
undefined Image 7: Lysine Acetyltransferase Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using CBP (D6C5) Rabbit mAb.
undefined Image 8: Lysine Acetyltransferase Antibody Sampler Kit
Confocal immunofluorescent analysis of HeLa cells using GCN5L2 (C26A10) Rabbit mAb (green). Actin filaments have been labeled with DY554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).
undefined Image 9: Lysine Acetyltransferase 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 1

Cell Signaling Technology Logo
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 µM) for 1 hour and PCAF (C14G9) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 9 (upper), including NR4A3 (lower), a known target gene of PCAF (see additional figure containing ChIP-qPCR data).
undefined Image 10: Lysine Acetyltransferase Antibody Sampler Kit
Western blot analysis of hypo- or hyper-acetylated recombinant p300 HAT domains, either wild-type or K1499R mutant, using Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody (upper). Also shown in the corresponding coomassie stained SDS-PAGE gel (lower). (Details are described in Thompson, P.A. et al. (2004) Nat. Struct. Mol. Biol. 11, 308-315.)
undefined Image 11: Lysine Acetyltransferase Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 μM, 1h) and CBP (D6C5) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using 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 CBP (see additional figure containing ChIP-qPCR data).
undefined Image 12: Lysine Acetyltransferase 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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using CBP (D6C5) Rabbit mAb.
undefined Image 13: Lysine Acetyltransferase Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human adenocarcinoma of the colon using CBP (D6C5) Rabbit mAb.
undefined Image 14: Lysine Acetyltransferase Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 µM) and either PCAF (C14G9) Rabbit mAb or Normal Rabbit IgG #2729, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. 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 15: Lysine Acetyltransferase 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 1

Cell Signaling Technology Logo
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells, treated with Forskolin #3828 (30 μM, 1h) and either CBP (D6C5) 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 16: Lysine Acetyltransferase Antibody Sampler Kit
Confocal immunofluorescent analysis of HeLa cells using CBP (D6C5) Rabbit mAb (green) and DyLight™ 554 Phalloidin #13054 (red).
undefined Image 17: Lysine Acetyltransferase Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human gastric adenocarcinoma using CBP (D6C5) Rabbit mAb.
undefined Image 18: Lysine Acetyltransferase 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 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded mouse ovary using CBP (D6C5) Rabbit mAb.
undefined Image 19: Lysine Acetyltransferase Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded normal human esophagus using CBP (D6C5) Rabbit mAb.
undefined Image 20: Lysine Acetyltransferase Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded LL2 syngeneic tumor using CBP (D6C5) Rabbit mAb.
undefined Image 21: Lysine Acetyltransferase 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 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded mouse colon using CBP (D6C5) Rabbit mAb.
undefined Image 22: Lysine Acetyltransferase Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded mouse cortex using CBP (D6C5) Rabbit mAb.
undefined Image 23: Lysine Acetyltransferase Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded mouse ovary using CBP (D6C5) Rabbit mAb.
undefined Image 24: Lysine Acetyltransferase 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 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human adenoid cystic carcinoma of the salivary gland using CBP (D6C5) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
undefined Image 25: Lysine Acetyltransferase Antibody Sampler Kit
Immunoprecipitation of CBP from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or CBP (D6C5) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using CBP (D6C5) Rabbit mAb.
undefined Image 26: Lysine Acetyltransferase Antibody Sampler Kit
Simple Western™ analysis of lysates (1 mg/mL) from A431 cells using Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody #4771. 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 27: Lysine Acetyltransferase 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 1

Cell Signaling Technology Logo
Simple Western™ analysis of lysates (0.1 mg/mL) from 293T cells using CBP (D6C5) Rabbit mAb #7389. 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 Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66 - 440 kDa separation module.
undefined Image 28: Lysine Acetyltransferase 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.