Revision 2

#56062

Store at -20C

Senescence Marker Antibody Sampler Kit

1 Kit

(8 x 20 microliters)

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

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

Product Includes Product # Quantity Mol. Wt Isotype/Source
p16 INK4A (D3W8G) Rabbit Monoclonal Antibody9280320 µl16 kDaRabbit IgG
p21 Waf1/Cip1 (12D1) Rabbit Monoclonal Antibody294720 µl21 kDaRabbit IgG
Phospho-Histone H2A.X (Ser139) (20E3) Rabbit Monoclonal Antibody971820 µl15 kDaRabbit IgG
Lamin B1 (D9V6H) Rabbit Monoclonal Antibody1343520 µl68, 45 kDaRabbit IgG
HMGB1 (D3E5) Rabbit Monoclonal Antibody689320 µl29 kDaRabbit IgG
IL-6 (D3K2N) Rabbit Monoclonal Antibody1215320 µl21-28 kDaRabbit IgG
TNF-alpha (D5G9) Rabbit Monoclonal Antibody694520 µl18, 25 kDaRabbit IgG
MMP-3 (D7F5B) Rabbit Monoclonal Antibody1435120 µl60 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 Senescence Marker Antibody Sampler Kit provides an economical means of detecting multiple markers of cellular senescence. The kit includes enough antibody to perform 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 antibody.

Background

Senescence is characterized by stable stress-induced proliferative arrest and resistance to mitogenic stimuli, as well as the secretion of proteins such as cytokines, growth factors and proteases. These secreted proteins comprise the senescence-associated secretory phenotype (SASP). Senescent cells are thought to accumulate as an organism ages, and contribute to age-related diseases, including cancer, through promotion of inflammation and disruption of normal cellular function (1,2).
Because there is no single biomarker that can be used to definitively identify senescent cells, researchers must rely on a collection of biomarkers commonly associated with senescence. The Senescence Marker Antibody Sampler Kit provides a collection of antibodies to commonly used biomarkers of senescence-associated cell cycle arrest (p16 INK4A, p21 Waf1/Cip1), senescence-associated DNA damage (gamma-Histone H2A.X), and the SASP (HMGB1, IL-6, TNF-alpha, MMP3). The kit also includes an antibody to Lamin B1, which is frequently reduced in senescent cells.

Trademarks and Patents

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

U.S. Patent No. 5,675,063.

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 1 ng recombinant Human Interleukin-6 (hIL-6) #8904 using IL-6 (D3K2N) Rabbit mAb.
undefined Image 1: Senescence Marker Antibody Sampler Kit
Western blot analysis of extracts from HeLa cells (lane 1) or LMNB1 knock-out cells (lane 2) using Lamin B1 (D9V6H) Rabbit mAb #13435 (upper), and β-actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the LMNB1 knock-out HeLa cells confirms specificity of the antibody for LMNB1.
undefined Image 2: Senescence Marker Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using MMP-3 (D7F5B) Rabbit mAb.
undefined Image 3: Senescence Marker 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 from control HeLa cells (lane 1) or p21 Waf1/Cip1 knockout HeLa cells (lane 2) using p21 Waf1/Cip1 (12D1) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the p21 Waf1/Cip1 knockout HeLa cells confirms specificity of the antibody for p21 Waf1/Cip1.
undefined Image 4: Senescence Marker Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using HMGB1 (D3E5) Rabbit mAb.
undefined Image 5: Senescence Marker Antibody Sampler Kit
Western blot analysis of extracts from the media of THP-1 cells differentiated with TPA #4174 (80 nM; overnight), with or without LPS (1 μg/ml; overnight), using TNF-α (D5G9) Rabbit mAb.
undefined Image 6: Senescence Marker 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
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 7: Senescence Marker Antibody Sampler Kit
Western blot analysis of extracts from HeLa and HUVEC cells using p16 INK4A (D3W8G) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
undefined Image 8: Senescence Marker Antibody Sampler Kit
Western blot analysis of extracts from untreated or UV-treated 293 cells, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb (upper) or Histone H2A.X Antibody #2595 (lower).
undefined Image 9: Senescence Marker 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 THP-1 cells differentiated with TPA #4174 (80 nM, overnight), untreated (-) or LPS-treated (100 ng/ml, 6 hr; +) and treated with Brefeldin A #9972 (300 ng/mL, last 3 hr of stimulation; +), using IL-6 (D3K2N) Rabbit mAb or β-Actin (D6A8) Rabbit mAb #8457 (lower).
undefined Image 10: Senescence Marker Antibody Sampler Kit
Western blot analysis of extracts from various cell lines and tissues using Lamin B1 (D9V6H) Rabbit mAb.
undefined Image 11: Senescence Marker Antibody Sampler Kit
Western blot analysis of extracts from various cell types using p21 Waf1/Cip1 (12D1) Rabbit mAb.
undefined Image 12: Senescence Marker 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
Immunohistochemical analysis of paraffin-embedded mouse lung using HMGB1 (D3E5) Rabbit mAb.
undefined Image 13: Senescence Marker Antibody Sampler Kit
Western blot analysis of extracts from the media of mouse bone marrow derived macrophages (mBMDM), untreated (-) or treated with Lipopolysaccharides (LPS) #14011 (50 ng/ml, 4 hr; +) followed by Nigericin (15 μM, 45 min; +), using HMGB1 (D3E5) Rabbit mAb.
undefined Image 14: Senescence Marker Antibody Sampler Kit
Western blot analysis of extracts from THP-1 cells differentiated with TPA #4174 (80 nM; overnight), with or without LPS (1 μg/ml; various time points), using TNF-α (D5G9) Rabbit mAb.
undefined Image 15: Senescence Marker 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
Immunoprecipitation of p16 INK4A from HeLa cells. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is p16 INK4A (D3W8G) Rabbit mAb. Western blot was performed using p16 INK4A (D3W8G) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
undefined Image 16: Senescence Marker Antibody Sampler Kit
Immunoprecipitation of IL-6 from NCI-H460 cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or IL-6 (D3K2N) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using IL-6 (D3K2N) Rabbit mAb.
undefined Image 17: Senescence Marker Antibody Sampler Kit
Western blot analysis of extracts from HeLa cells, untreated (-) or treated with Staurosporine #9953 (1 μM, 3 hr; +), using Lamin B1 (D9V6H) Rabbit mAb.
undefined Image 18: Senescence Marker 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 HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® p21 Waf1/Cip1 siRNA II (+), using p21 Waf1/Cip1 (12D1) Rabbit mAb #2947 and α-Tubulin (11H10) Rabbit mAb #2125. The p21 Waf1/Cip1 (12D1) Rabbit mAb confirms silencing of p21 Waf1/Cip1 expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of p21 Waf1/Cip1 siRNA.
undefined Image 19: Senescence Marker Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human colon carcioma using HMGB1 (D3E5) Rabbit mAb.
undefined Image 20: Senescence Marker Antibody Sampler Kit
Western blot analysis of recombinant human TNF-α using TNF-α (D5G9) Rabbit mAb (left) or TNF-α Antibody #3707 (right).
undefined Image 21: Senescence Marker 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
Immunoprecipitation of p21 from human umbillical vein endothelial cells (HUVECs) using p21 Waf1/Cip1 (12D1) Rabbit mAb. Western blot detection was performed using the same antibody.
undefined Image 22: Senescence Marker Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human lung carcioma using HMGB1 (D3E5) Rabbit mAb.
undefined Image 23: Senescence Marker Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using p21 Waf1/Cip1 (12D1) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
undefined Image 24: Senescence Marker 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
Immunohistochemical analysis of paraffin-embedded human prostate carcioma using HMGB1 (D3E5) Rabbit mAb.
undefined Image 25: Senescence Marker Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded HeLa cells, transfected with SignalSilence® Control siRNA (Unconjugated) #6568 (left) or SignalSilence® p21 Waf1/Cip1 siRNA II #6558 (right), using p21 Waf1/Cip1 (12D1) Rabbit mAb.
undefined Image 26: Senescence Marker Antibody Sampler Kit
Confocal immunofluorescent analysis of MCF7 cells using p21 Waf1/Cip1 (12D1) Rabbit mAb (red) and Phospho-Histone H3 (Ser10) (6G3) Mouse mAb #9706 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 27: Senescence Marker 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
Flow cytometric analysis of Daudi cells using p21 Waf1/Cip1 (12D1) Rabbit mAb (right) and Propidium Iodide (PI)/RNase Staining Solution #4087, compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
undefined Image 28: Senescence Marker Antibody Sampler Kit
Simple Western™ analysis of lysates (0.1 mg/mL) from NK-92 cells treated with TPA + lonomycin + BFA using IL-6 (D3K2N) Rabbit mAb #12153. 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 2-40 kDa separation module.
undefined Image 29: Senescence Marker Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded HT-29 cells untreated (left) or UV-treated (right), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb.
undefined Image 30: Senescence Marker 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
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb.
undefined Image 31: Senescence Marker Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb, showing nuclear localization.
undefined Image 32: Senescence Marker Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human lung carcinoma untreated (left) or lambda-phosphatase-treated (right), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb.
undefined Image 33: Senescence Marker 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 HeLa cells, untreated (left) or UV-treated (right), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
undefined Image 34: Senescence Marker Antibody Sampler Kit
Flow cytometric analysis of HeLa cells, untreated (blue) or treated with UV (100 mJ, 2hr recovery; green) using Phospho-H2A.X (Ser139) (20E3) 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 35: Senescence Marker Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human ovarian clear cell carcinoma using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb performed on the Leica BOND Rx.
undefined Image 36: Senescence Marker 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
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb performed on the Leica BOND Rx.
undefined Image 37: Senescence Marker Antibody Sampler Kit
Confocal immunofluorescent analysis of fixed frozen mouse colon at low magnification (left) and high magnification (right) using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit Monoclonal Antibody (green). After blocking free secondary antibody binding sites with Rabbit (DA1E) Monoclonal Antibody IgG Isotype Control #3900, the tissue was then labeled using CD45 (D3F8Q) Rabbit Monoclonal Antibody (Alexa Fluor® 555 Conjugate) #19581 (red), beta-Catenin (D10A8) Rabbit Monoclonal Antibody (Alexa Fluor® 647 Conjugate) #23371 (yellow pseudocolor), and ProLong Gold Antifade Reagent with DAPI #8961 (blue).
undefined Image 38: Senescence Marker Antibody Sampler Kit

Confocal immunofluorescent analysis of fixed frozen mouse thymus using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit Monoclonal Antibody (green). After blocking free secondary antibody binding sites with Rabbit (DA1E) Monoclonal Antibody IgG Isotype Control #3900, the tissue was then labeled using CD45 (D3F8Q) Rabbit Monoclonal Antibody (Alexa Fluor® 555 Conjugate) #19581 (red) and beta-Catenin (D10A8) Rabbit Monoclonal Antibody (Alexa Fluor® 647 Conjugate) #23371 (cyan pseudocolor).

undefined Image 39: Senescence Marker 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.