Revision 1

#11843Store at -20C

1 Kit

(4 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
MEK1/2 (D1A5) Rabbit mAb 8727 20 µl 45 kDa Rabbit IgG
AIF (D39D2) XP® Rabbit mAb 5318 20 µl 67 kDa Rabbit IgG
Histone H3 (D1H2) XP® Rabbit mAb 4499 20 µl 17 kDa Rabbit IgG
Vimentin (D21H3) XP® Rabbit mAb 5741 20 µl 57 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

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

Description

The Cell Fractionation Antibody Sampler Kit provides an economical means for determining the purity of each distinctly separated subcellular fraction by western blot using Cell Signaling Technology's Cell Fractionation Kit #9038. This antibody sampler kit includes enough primary antibody to perform at least two western blots per 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 antibody.

Background

Knowledge of the subcellular location of a protein may reveal the potential role it plays in a variety of cellular processes. Antibodies in the Cell Fractionation Antibody Sampler Kit can be used as a marker to ensure that each subcellular fraction is efficiently separated from the next. MEK1 and MEK2, also called MAPK or Erk Kinases, are dual-specificity protein kinases that function in a mitogen activated protein kinase cascade (1). Apoptosis-inducing factor (AIF) is a ubiquitously expressed flavoprotein that plays a critical role in caspase-independent apoptosis (2). Core histone protein, H3 is one of the primary building blocks of chromatin that makes up the nucleosome (3). Vimentin is a cell-specific intermediate filament with mesenchyme origin that contributes to making up of the cytoskeleton (4). MEK1/2, AIF, histone H3, and vimentin localize to the cytoplasm, mitochondria, nucleus, and cytoskeleton, respectively.

  1. Crews, C.M. et al. (1992) Science 258, 478-80.
  2. Lipton, S.A. and Bossy-Wetzel, E. (2002) Cell 111, 147-50.
  3. Workman, J.L. and Kingston, R.E. (1998) Annu Rev Biochem 67, 545-79.
  4. Eng, L.F. et al. (2000) Neurochem Res 25, 1439-51.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a 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.

    Revision 1
    #11843

    Cell Fractionation Antibody Sampler Kit

    Cell Fractionation Antibody Sampler Kit: Image 1 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse kidney labeled with Vimentin (D21H3) XP® Rabbit mAb (left, green) and co-labeled with F4/80 (BM8.1) Rat mAb (right, red), and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
    Cell Fractionation Antibody Sampler Kit: Image 2 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse colon labeled with Vimentin (D21H3) XP® Rabbit mAb (left, green) and co-labeled with F4/80 (BM8.1) Rat mAb (right, red), and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
    Cell Fractionation Antibody Sampler Kit: Image 3 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse cerebellum labeled with Vimentin (D21H3) XP® Rabbit mAb (left, green) and co-labeled with F4/80 (BM8.1) Rat mAb (right, red), and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
    Cell Fractionation Antibody Sampler Kit: Image 4 Expand Image
    Simple Western™ analysis of lysates (1 mg/mL) from HeLa cells using Vimentin (D21H3) XP ® Rabbit mAb #5741. The virtual lane view (left) shows a single 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.
    Cell Fractionation Antibody Sampler Kit: Image 5 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse cerebellum labeled with Vimentin (D21H3) XP® Rabbit mAb (left, green) and co-labeled with F4/80 (BM8.1) Rat mAb (right, red), and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
    Cell Fractionation Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from various cell lines using Histone H3 (D1H2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 7 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Histone H3 (D1H2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from Jurkat, C2C12, and C6 cells using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 9 Expand Image
    Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from various cell lines using Vimentin (D21H3) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 11 Expand Image
    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.
    Cell Fractionation Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from various cell lines using MEK1/2 (D1A5) Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 13 Expand Image
    Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic mammary tumor using Histone H3 (D1H2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 14 Expand Image
    Immunohistochemical analysis of paraffin-embedded human papillary thyroid carcinoma using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 15 Expand Image
    Western blot analysis of extracts from control HeLa cells (lane 1) or Vimentin knockout HeLa cells (lane 2) using Vimentin (D21H3) XP® Rabbit mAb #5741 (upper) or β-Actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the Vimentin knockout HeLa cells confirms specificity of the antibody for Vimentin.
    Cell Fractionation Antibody Sampler Kit: Image 16 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using MEK1/2 (D1A5) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Cell Fractionation Antibody Sampler Kit: Image 17 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using Histone H3 (D1H2) XP® Rabbit mAb (green) and β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red).
    Cell Fractionation Antibody Sampler Kit: Image 18 Expand Image
    Immunohistochemical analysis of paraffin-embedded LL/2 syngeneic tumor using Histone H3 (D1H2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 19 Expand Image
    Confocal immunofluorescent analysis of mouse brainstem using AIF (D39D2) XP® Rabbit mAb #5318 (green) and GFAP (GA5) Mouse mAb #3670 (red). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Cell Fractionation Antibody Sampler Kit: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 21 Expand Image
    Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using Vimentin (D21H3) XP® Rabbit mAb performed on the Leica® BOND Rx.
    Cell Fractionation Antibody Sampler Kit: Image 22 Expand Image
    Flow cytometric analysis of Jurkat cells using MEK1/2 (D1A5) 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.
    Cell Fractionation Antibody Sampler Kit: Image 23 Expand Image
    Flow cytometric analysis of HeLa cells using Histone H3 (D1H2) XP® Rabbit mAb (solid line) or a 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.
    Cell Fractionation Antibody Sampler Kit: Image 24 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain using Histone H3 (D1H2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 25 Expand Image
    Confocal immunofluorescent analysis of mouse cerebellum using AIF (D39D2) XP® Rabbit mAb #5318 (green), GFAP (GA5) Mouse mAb #3670 (red), and F4/80 (BM8.1) Rat mAb #71299 (blue).
    Cell Fractionation Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 27 Expand Image
    Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using Vimentin (D21H3) XP® Rabbit mAb performed on the Leica® BOND Rx.
    Cell Fractionation Antibody Sampler Kit: Image 28 Expand Image
    Immunohistochemical analysis of paraffin-embedded rhesus monkey liver using Histone H3 (D1H2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 29 Expand Image
    Confocal immunofluorescent analysis of mouse striatum using AIF (D39D2) XP® Rabbit mAb #5318 (green), GFAP (GA5) Mouse mAb #3670 (red), and F4/80 (BM8.1) Rat mAb #71299 (blue).
    Cell Fractionation Antibody Sampler Kit: Image 30 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 31 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Vimentin (D21H3) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 32 Expand Image
    Confocal immunofluorescent analysis of mouse thalamus using AIF (D39D2) XP® Rabbit mAb #5318 (green), GFAP (GA5) Mouse mAb #3670 (red). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Cell Fractionation Antibody Sampler Kit: Image 33 Expand Image
    Immunohistochemical analysis of paraffin-embedded human papillary carcinoma of the breast using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 34 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse colon using Vimentin (D21H3) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 35 Expand Image
    Confocal immunofluorescent analysis of C2C12 cells using AIF (D39D2) XP® Rabbit mAb #5318 (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Cell Fractionation Antibody Sampler Kit: Image 36 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human testis using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 37 Expand Image
    Immunohistochemical analysis of paraffin-embedded rat colon using Vimentin (D21H3) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 38 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using AIF (D39D2) XP Rabbit mAb (green), showing colocalization with mitochondria that have been labeled with MitoTracker® Red CMXRos (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Cell Fractionation Antibody Sampler Kit: Image 39 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human spleen using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 40 Expand Image
    Confocal immunofluorescent analysis of SNB19 cells using Vimentin (D21H3) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Cell Fractionation Antibody Sampler Kit: Image 41 Expand Image
    Immunohistochemical analysis of paraffin-embedded rhesus kidney using Vimentin (D21H3) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 42 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human skeletal muscle using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 43 Expand Image
    Flow cytometric analysis of MCF7 cells (blue, negative) and HeLa cells (green, positive) using Vimentin (D21H3) XP® Rabbit mAb(solid lines) or a 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.
    Cell Fractionation Antibody Sampler Kit: Image 44 Expand Image
    Immunohistochemical analysis of paraffin-embedded Syrian hamster small intestine using Vimentin (D21H3) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 45 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human liver using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 46 Expand Image
    Immunohistochemical analysis of paraffin-embedded human tonsil using Vimentin (D21H3) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
    Cell Fractionation Antibody Sampler Kit: Image 47 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human kidney using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 48 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human breast using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 49 Expand Image
    Immunohistochemical analysis of paraffin-embedded A20 syngeneic tumor using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 50 Expand Image
    Immunohistochemical analysis of paraffin-embedded LL/2 syngeneic tumor using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 51 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse testis using AIF (D39D2) XP® Rabbit mAb.
    Cell Fractionation Antibody Sampler Kit: Image 52 Expand Image
    Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using AIF (D39D2) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).