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78551
Mouse Reactive Senescence Marker Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Mouse Reactive Senescence Marker Antibody Sampler Kit #78551

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Simple Western™ analysis of lysates (1.0 mg/mL) from 3T3-L1 cells using MMP-2 (D2O4T) Rabbit mAb #87809. 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 12-230 kDa separation module.

Western blot analysis of extracts from Raw 264.7 cells, untreated (-) or treated (+) with LPS (100 ng/mL, 6 hr) and Brefeldin A #9972 (300 ng/mL, last 3 hr of stimulation), using TNF-α (D2D4) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from Raw 264.7 cells, untreated (-) or treated with LPS (100 ng/ml, 6 hr) and Brefeldin A #9972 (300 ng/ml, last 3 hr of stimulation; +), using IL-6 (D5W4V) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from various cell lines and rat spleen tissue using Lamin B1 (E6M5T) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Relatively low expression of Lamin B1 protein in U-87 MG cells is consistent with the predicted expression pattern.
Immunoprecipitation of p16 INK4A protein from mouse A20 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is p16 INK4A (E5F3Y) Rabbit mAb. Western blot analysis was performed using p16 INK4A (E5F3Y) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody. 
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing full-length mouse p16 INK4A protein (mINK4A-Myc/DDK; +) or full-length mouse p15 INK4B protein (mINK4B-Myc/DDK; +), using p16 INK4A (E5F3Y) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western blot analysis of extracts from MEF cells, untreated (-) or treated with Doxorubicin #5927 (250 nM, 24 hours followed by 7 days without doxorubicin; +), using p16 INK4A (E5F3Y) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). p16 INK4A protein expression is induced by doxorubicin as expected.
Western blot analysis of extracts from various mouse cell lines using p16 INK4A (E5F3Y) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Negative expression of p16 INK4A protein in NIH/3T3 cells is consistent with the predicted expression pattern.
Western blot analysis of extracts from control C2C12 cells (lane 1) or p21 Waf1/Cip1 knockout C2C12 cells (lane 2) using p21 Waf1/Cip1 (E2R7A) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The absence of signal in the p21 Waf1/Cip1 knockout C2C12 cells confirms specificity of the antibody for p21 Waf1/Cip1.
Western blot analysis of extracts from various cell lines using HMGB1 (D3E5) Rabbit mAb.
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.
Western blot analysis of extracts from U87, 3T3-L1 and C2C12 cells using MMP-2 (D4O4T) Rabbit mAb.
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).
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.
Western blot analysis of 1 ng recombinant mouse TNF-α #5178 using TNF-α (D2D4) XP® Rabbit mAb.
Western blot analysis of recombinant Mouse His6Interleukin-6 (mHis6IL-6) using IL-6 (D5W4V) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded normal human breast using Lamin B1 (E6M5T) Rabbit mAb.
Western blot analysis of extracts from C2C12, NIH/3T3 (mouse), and MCF7 (human) cells, untreated (-) or treated with Nutlin 3a (10 μM, 24 hr; +), using p21 Waf1/Cip1 (E2R7A) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Immunohistochemical analysis of paraffin-embedded mouse lung using HMGB1 (D3E5) Rabbit mAb.
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.
Immunoprecipitation of MMP-2 protein from U87 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is MMP-2 (D2O4T) Rabbit mAb. Western blot analysis was performed using MMP-2 (D2O4T) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb performed on the Leica BOND Rx.
Immunoprecipitation of TNF-α from Raw 264.7 cell extracts, treated with LPS (100 ng/mL, 6 hr) and Brefeldin A #9972 (300 ng/mL, last 3 hr of stimulation), using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or TNF-α (D2D4) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using TNF-α (D2D4) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) #3678 and Anti-mouse IgG, HRP-linked Antibody #7076 were used as sencondary antibodies.
Immunoprecipitation of IL-6 from Raw 264.7 cell extracts treated with LPS (100 ng/ml, 6 hr) and Brefeldin A #9972 (300 ng/ml, last 3 hr of stimulation) using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or IL-6 (D5W4V) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using IL-6 (D5W4V) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded BeWo cell pellet (left, high-expressing) or U-87 MG cell pellet (right, low-expressing) using Lamin B1 (E6M5T) Rabbit mAb.
Immunoprecipitation of p21 Waf1/Cip1 protein from extracts of C2C12 cells treated with Nutlin 3a (10 μM, 24 hr).  Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is p21 Waf1/Cip1 (E2R7A) Rabbit mAb. Western blot analysis was performed using  p21 Waf1/Cip1 (E2R7A) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody. 
Immunohistochemical analysis of paraffin-embedded human colon carcioma using HMGB1 (D3E5) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb, showing nuclear localization.
Confocal immunofluorescent analysis of Raw 264.7 cells, treated with LPS (100 ng/mL, 6 hr; left) or untreated (right), using TNF-α (D2D4) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent analysis of Raw 264.7 cells, untreated (left) or treated with LPS (100 ng/ml, 6 hr; right), using IL-6 (D5W4V) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor= DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded human esophageal adenocarcinoma using Lamin B1 (E6M5T) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcioma using HMGB1 (D3E5) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb.
Flow cytometric analysis of Raw 264.7 cells, untreated (blue) or treated with LPS (100 ng/ml, 6 hr) and Brefeldin A #9972 (300 ng/ml, last 3 hr of treatment) (green), using TNF-α (D2D4) XP® 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.
Flow cytometric analysis of Raw 264.7 cells, treated with LPS (100 ng/ml, 6 hr) and Brefeldin A #9972 (300 ng/ml, last 3 hr of LPS treatment) (green) or Brefeldin A alone (300 ng/ml, 3 hr) (blue) using IL-6 (D5W4V) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.
Immunohistochemical analysis of paraffin-embedded human squamous cell carcinoma of the tonsil using Lamin B1 (E6M5T) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human prostate carcioma using HMGB1 (D3E5) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma untreated (left) or lambda-phosphatase-treated (right), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using Lamin B1 (E6M5T) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using Lamin B1 (E6M5T) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded HT-29 cells untreated (left) or UV-treated (right), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb.
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).
Immunohistochemical analysis of paraffin-embedded human mature ovarian teratoma using Lamin B1 (E6M5T) Rabbit mAb.
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.
Immunohistochemical analysis of paraffin-embedded human ovarian clear cell carcinoma using Lamin B1 (E6M5T) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemical analysis of paraffin-embedded normal human pancreas using Lamin B1 (E6M5T) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded normal human prostate using Lamin B1 (E6M5T) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded normal human thymus using Lamin B1 (E6M5T) Rabbit mAb.
To Purchase # 78551
Cat. # Size Qty. Price
78551T
1 Kit  (6 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
p16 INK4A (E5F3Y) Rabbit mAb 29271 20 µl
  • WB
  • IP
M 16 Rabbit IgG
p21 Waf1/Cip1 (E2R7A) Rabbit mAb 37543 20 µl
  • WB
  • IP
H M 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 (E6M5T) Rabbit mAb 17416 20 µl
  • WB
  • IHC
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 (D5W4V) XP® Rabbit mAb 12912 20 µl
  • WB
  • IP
  • IF
  • F
M 24 Rabbit IgG
TNF-α (D2D4) XP® Rabbit mAb 11948 20 µl
  • WB
  • IP
  • IF
  • F
M 17,25,28 Rabbit IgG
MMP-2 (D2O4T) Rabbit mAb 87809 20 µl
  • WB
  • IP
H M 64,72 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Mouse Reactive Senescence Marker Antibody Sampler Kit provides an economical means of detecting multiple markers of cellular senescence in mouse samples. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Mouse Reactive Senescence Marker Antibody Sampler Kit detects endogenous levels of its target protein. Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb detects endogenous levels of H2A.X only when phosphorylated at Ser139.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Pro67 of mouse p16 INK4A protein, Arg455 of human lamin B1 protein, Ala137 of human HMGB1 protein, Pro117 of human MMP-2 protein, a synthetic phosphopeptide corresponding to residues surrounding Ser139 of human H2A.X protein, or recombinant proteins specific to human p21 Waf1/Cip1 protein or mouse TNF-α 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 (3). The Mouse Reactive 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-α, MMP2). The kit also includes an antibody to lamin B1, which is frequently reduced in senescent cells.

Pathways

Explore pathways related to this product.

Limited Uses

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