Revision 1

#56062Store at -20C

1 Kit

(8 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
p16 INK4A (D3W8G) Rabbit mAb 92803 20 µl 16 kDa Rabbit IgG
p21 Waf1/Cip1 (12D1) Rabbit mAb 2947 20 µl 21 kDa Rabbit IgG
Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb 9718 20 µl 15 kDa Rabbit IgG
Lamin B1 (D9V6H) Rabbit mAb 13435 20 µl 68, 45 kDa Rabbit IgG
HMGB1 (D3E5) Rabbit mAb 6893 20 µl 29 kDa Rabbit IgG
IL-6 (D3K2N) Rabbit mAb 12153 20 µl 21-28 kDa Rabbit IgG
TNF-α (D5G9) Rabbit mAb 6945 20 µl 18, 25 kDa Rabbit IgG
MMP-3 (D7F5B) Rabbit mAb 14351 20 µl 60 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 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.

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.

Background References

    Trademarks and Patents

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

    使用に関する制限

    法的な権限を与えられたCSTの担当者が署名した書面によって別途明示的に合意された場合を除き、 CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、 または、ここに定められた条件と異なるものは、法的な権限を与えられたCSTの担当者が別途書面にて受諾した場合を除き、拒絶され、 いかなる効力も効果も有しません。

    研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

    Revision 1
    #56062

    Senescence Marker Antibody Sampler Kit

    Senescence Marker Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™ analysis of lysates (1.0 mg/mL) from MCF-7 cells using p21 Waf1/Cip1 (12D1) Rabbit mAb #2947. 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.
    Senescence Marker Antibody Sampler Kit: Image 2 Expand Image
    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 3 Expand Image
    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 4 Expand Image
    Western blot analysis of extracts from various cell lines using MMP-3 (D7F5B) Rabbit mAb.
    Senescence Marker Antibody Sampler Kit: Image 5 Expand Image
    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 6 Expand Image
    Western blot analysis of extracts from various cell lines using HMGB1 (D3E5) Rabbit mAb.
    Senescence Marker Antibody Sampler Kit: Image 7 Expand Image
    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 8 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.
    Senescence Marker Antibody Sampler Kit: Image 9 Expand Image
    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 10 Expand Image
    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).
    Senescence Marker Antibody Sampler Kit: Image 11 Expand Image
    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.
    Senescence Marker Antibody Sampler Kit: Image 12 Expand Image
    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 13 Expand Image
    Western blot analysis of extracts from various cell lines and tissues using Lamin B1 (D9V6H) Rabbit mAb.
    Senescence Marker Antibody Sampler Kit: Image 14 Expand Image
    Western blot analysis of extracts from various cell types using p21 Waf1/Cip1 (12D1) Rabbit mAb.
    Senescence Marker Antibody Sampler Kit: Image 15 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse lung using HMGB1 (D3E5) Rabbit mAb.
    Senescence Marker Antibody Sampler Kit: Image 16 Expand Image
    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.
    Senescence Marker Antibody Sampler Kit: Image 17 Expand Image
    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 18 Expand Image
    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 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb performed on the Leica BOND Rx.
    Senescence Marker Antibody Sampler Kit: Image 20 Expand Image
    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 21 Expand Image
    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 22 Expand Image
    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 23 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcioma using HMGB1 (D3E5) Rabbit mAb.
    Senescence Marker Antibody Sampler Kit: Image 24 Expand Image
    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 25 Expand Image
    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 26 Expand Image
    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 27 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcioma using HMGB1 (D3E5) Rabbit mAb.
    Senescence Marker Antibody Sampler Kit: Image 28 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb.
    Senescence Marker Antibody Sampler Kit: Image 29 Expand Image
    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 30 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate carcioma using HMGB1 (D3E5) Rabbit mAb.
    Senescence Marker Antibody Sampler Kit: Image 31 Expand Image
    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 32 Expand Image
    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 33 Expand Image
    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 34 Expand Image
    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 35 Expand Image
    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 36 Expand Image
    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 37 Expand Image
    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.