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56062
Senescence Marker Antibody Sampler Kit
Primary Antibodies

Senescence Marker Antibody Sampler Kit #56062

Western Blotting Image 1

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).

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Western Blotting Image 2

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

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Western Blotting Image 3

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).

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Western Blotting Image 4

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

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Western Blotting Image 5

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

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Western Blotting Image 6

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

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Western Blotting Image 7

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.

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Western Blotting Image 8

Western blot analysis of extracts from various cell lines using MMP3 (D7F5B) Rabbit mAb.

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Western Blotting Image 9

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.

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IP Image 10

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.

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Western Blotting Image 11

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.

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IHC-P (paraffin) Image 12

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

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Western Blotting Image 13

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

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IHC-P (paraffin) Image 14

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

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Western Blotting Image 15

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).

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Western Blotting Image 16

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.

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IP Image 17

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.

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IHC-P (paraffin) Image 18

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).

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IHC-P (paraffin) Image 19

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

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IP Image 20

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.

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Western Blotting Image 21

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

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IHC-P (paraffin) Image 22

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).

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IHC-P (paraffin) Image 23

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

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IHC-P (paraffin) Image 24

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

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IHC-P (paraffin) Image 25

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.

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IHC-P (paraffin) Image 26

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

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IHC-P (paraffin) Image 27

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

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Flow Cytometry Image 28

Flow cytometric analysis of HeLa cells (red) and MCF7 cells (blue), using p21 Waf1/Cip1 (12D1) Rabbit mAb.

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IHC-P (paraffin) Image 29

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

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IF-IC Image 30

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).

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Flow Cytometry Image 31

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.

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IF-IC Image 32

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).

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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
MMP3 (D7F5B) Rabbit mAb 14351 20 µl
  • WB
H R 60 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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.

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

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.

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.
Entrez-Gene Id
3014 , 3146 , 3569 , 1029 , 4001 , 4314 , 1026 , 7124
Swiss-Prot Acc.
P16104 , P09429 , P05231 , P42771 , P20700 , P08254 , P38936 , P01375
For Research Use Only. Not For Use In Diagnostic Procedures.

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.

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