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56062
Senescence Marker Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Senescence Marker Antibody Sampler Kit #56062

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Senescence Marker Antibody Sampler Kit: Image 1

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.

Senescence Marker Antibody Sampler Kit: Image 2

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.

Senescence Marker Antibody Sampler Kit: Image 3

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.

Senescence Marker Antibody Sampler Kit: Image 4

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.

Senescence Marker Antibody Sampler Kit: Image 5

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.

Senescence Marker Antibody Sampler Kit: Image 6

Immunohistochemical analysis of paraffin-embedded mouse lung using HMGB1 (D3E5) Rabbit mAb.

Senescence Marker Antibody Sampler Kit: Image 7

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.

Senescence Marker Antibody Sampler Kit: Image 8

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.

Senescence Marker Antibody Sampler Kit: Image 9

Western blot analysis of extracts from various cell lines using MMP-3 (D7F5B) Rabbit mAb.

Senescence Marker Antibody Sampler Kit: Image 10

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.

Senescence Marker Antibody Sampler Kit: Image 11

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).

Senescence Marker Antibody Sampler Kit: Image 12

Western blot analysis of extracts from various cell types using p21 Waf1/Cip1 (12D1) Rabbit mAb.

Senescence Marker Antibody Sampler Kit: Image 13

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).

Senescence Marker Antibody Sampler Kit: Image 14

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).

Senescence Marker Antibody Sampler Kit: Image 15

Western blot analysis of extracts from various cell lines and tissues using Lamin B1 (D9V6H) Rabbit mAb.

Senescence Marker Antibody Sampler Kit: Image 16

Immunohistochemical analysis of paraffin-embedded human colon carcioma using HMGB1 (D3E5) Rabbit mAb.

Senescence Marker Antibody Sampler Kit: Image 17

Western blot analysis of 1 ng recombinant Human Interleukin-6 (hIL-6) #8904 using IL-6 (D3K2N) Rabbit mAb.

Senescence Marker Antibody Sampler Kit: Image 18

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.

Senescence Marker Antibody Sampler Kit: Image 19

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.

Senescence Marker Antibody Sampler Kit: Image 20

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).

Senescence Marker Antibody Sampler Kit: Image 21

Immunohistochemical analysis of paraffin-embedded HT-29 cells untreated (left) or UV-treated (right), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb.

Senescence Marker Antibody Sampler Kit: Image 22

Western blot analysis of extracts from HeLa cells, untreated (-) or treated with Staurosporine #9953 (1 μM, 3 hr; +), using Lamin B1 (D9V6H) Rabbit mAb.

Senescence Marker Antibody Sampler Kit: Image 23

Immunohistochemical analysis of paraffin-embedded human lung carcioma using HMGB1 (D3E5) Rabbit mAb.

Senescence Marker Antibody Sampler Kit: Image 24

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).

Senescence Marker Antibody Sampler Kit: Image 25

Western blot analysis of recombinant human TNF-α #8902 using TNF-α (D5G9) Rabbit mAb (left) or TNF-α Antibody #3707 (right).

Senescence Marker Antibody Sampler Kit: Image 26

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.

Senescence Marker Antibody Sampler Kit: Image 27

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb in the presence of control peptide (left) or Phospho-Histone H2A.X (Ser139) Blocking Peptide #1260 (right).

Senescence Marker Antibody Sampler Kit: Image 28

Immunohistochemical analysis of paraffin-embedded human prostate carcioma using HMGB1 (D3E5) Rabbit mAb.

Senescence Marker Antibody Sampler Kit: Image 29

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.

Senescence Marker Antibody Sampler Kit: Image 30

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb.

Senescence Marker Antibody Sampler Kit: Image 31

Western blot analysis of extracts from various cell lines using HMGB1 (D3E5) Rabbit mAb.

Senescence Marker Antibody Sampler Kit: Image 32

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb, showing nuclear localization.

Senescence Marker Antibody Sampler Kit: Image 33

Immunohistochemical analysis of paraffin-embedded human lung carcinoma untreated (left) or lambda-phosphatase-treated (right), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb.

Senescence Marker Antibody Sampler Kit: Image 34

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).

To Purchase # 56062T
Product # Size Price
56062T
1 Kit  (8 x 20 µl) $ 554

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
p16 INK4A (D3W8G) Rabbit mAb 92803 20 µl
  • WB
  • IP
H 16 Rabbit IgG
p21 Waf1/Cip1 (12D1) Rabbit mAb 2947 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H Mk 21 Rabbit IgG
Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb 9718 20 µl
  • WB
  • IHC
  • IF
  • F
H M R Mk 15 Rabbit IgG
Lamin B1 (D9V6H) Rabbit mAb 13435 20 µl
  • WB
  • IP
H M R 68, 45 Rabbit IgG
HMGB1 (D3E5) Rabbit mAb 6893 20 µl
  • WB
  • IHC
H M R Mk 29 Rabbit IgG
IL-6 (D3K2N) Rabbit mAb 12153 20 µl
  • WB
  • IP
H 21-28 Rabbit IgG
TNF-α (D5G9) Rabbit mAb 6945 20 µl
  • WB
  • IP
H 18, 25 Rabbit IgG
MMP-3 (D7F5B) Rabbit mAb 14351 20 µl
  • WB
H R 60 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product 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.

Specificity / Sensitivity

Each antibody in the Senescence Marker Antibody Sampler Kit detects endogenous levels of its target protein.

Source / Purification

Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to residues surrounding Ala34 of human p16 INK4A protein, residues near the carboxy-terminus of human p21, residues surrounding Ser139 of human histone H2A.X protein, residues surrounding Lys415 of human lamin B1 protein, residues surrounding Ala137 of human HMGB1 protein, recombinant human IL-6 protein, recombinant human TNF-α protein, and residues surrounding Ser417 of human MMP3 protein.

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.

  1. Tchkonia, T. et al. (2013) J Clin Invest 123, 966-72.
  2. Sun, Y. et al. (2018) Trends Mol Med 24, 871-885.

Pathways & Proteins

Explore pathways + proteins related to this product.

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's 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's 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.

For Research Use Only. Not For Use In Diagnostic Procedures.
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U.S. Patent No. 5,675,063.