Revision 1
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
IL-1β (3A6) Mouse mAb 12242 20 µl 17,31 kDa Mouse IgG1
CCL5/RANTES Antibody 2989 20 µl 10 kDa Rabbit 
PAI-1 Antibody 27535 20 µl 50 kDa Rabbit 
TNF-α (D2D4) XP® Rabbit mAb 11948 20 µl 17,25,28 kDa Rabbit IgG
MCP-1 (E9R7Z) Rabbit mAb 41987 20 µl 11-13, 20-30 kDa Rabbit IgG
MMP-2 (D2O4T) Rabbit mAb 87809 20 µl 64,72 kDa Rabbit IgG
IL-6 (D5W4V) XP® Rabbit mAb 12912 20 µl 24 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl Horse 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit provides an economical means of detecting multiple components of the SASP in mouse samples. The kit includes enough antibodies 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 antibodies.

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). The composition of the SASP varies, and SASP components can be either beneficial or deleterious in human disease, depending on the context (3).
The Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit provides a collection of antibodies to various SASP components, including TNF-α, interleukin-6 (IL-6), the multifunctional cytokine IL-1β, the chemokines RANTES/CCL5 and MCP-1, the matrix metalloprotease MMP-2, and the serine-protease inhibitor PAI-1.

  1. Tchkonia, T. et al. (2013) J Clin Invest 123, 966-72.
  2. Sun, Y. et al. (2018) Trends Mol Med 24, 871-885.
  3. Rao, S.G. and Jackson, J.G. (2016) Trends Cancer 2, 676-687.

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.
    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 1
    #85741

    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit

    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™ analysis of lysates (1.0 mg/mL) from THP-1 cells treated with TPA (80nM, O/N) and LPS (1ug/ml, 1 hr) using IL-1β (3A6) Mouse mAb #12242. 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.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 2 Expand Image

    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.

    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 3 Expand Image
    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).
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from THP-1 cells, untreated (-) or LPS-treated (100 ng/mL, 3 hr; +), using IL-1β (3A6) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 5 Expand Image
    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).
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from mouse placenta and mouse spleen using PAI-1 Antibody (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The differential in PAI-1 expression between placenta and spleen is consistent with their reported RNA expression profiles (www.proteinatlas.org), confirming specificity of the antibody for PAI-1.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of HeLa cells, mock transfected or transfected with mouse CCL5/RANTES, using CCL5/RANTES Antibody.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from RAW 264.7 cells, untreated (-) or treated with LPS #14011 (1 μg/ml, overnight; +), using MCP-1 (E9R7Z) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 9 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.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 10 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.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from U87, 3T3-L1 and C2C12 cells using MMP-2 (D4O4T) Rabbit mAb.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of 1 ng recombinant mouse TNF-α #5178 using TNF-α (D2D4) XP® Rabbit mAb.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from Raw 264.7 cells, untreated (-) or treated with Brefeldin A #9972 (300 ng/mL, last 3 hr of stimulation; +) and LPS (100 ng/mL, 6 hr; +), using IL-1β (3A6) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 14 Expand Image
    Western blot analysis of recombinant Mouse His6Interleukin-6 (mHis6IL-6) using IL-6 (D5W4V) XP® Rabbit mAb.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 15 Expand Image
    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.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 16 Expand Image
    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.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 17 Expand Image
    Western blot analysis of 1 ng recombinant Human Interleukin-1β (hIL-1β) #8900 and 1 ng recombinant Mouse Interleukin-1β (mIL-1β) #5204 using IL-1β (3A6) Mouse mAb.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 18 Expand Image
    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.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 19 Expand Image
    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).
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded human large intestine (chronic colitis of the colon) using IL-1-β (3A6) Mouse mAb.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 21 Expand Image
    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).
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 22 Expand Image
    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.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 23 Expand Image
    Immunohistochemical analysis of paraffin-embedded human large intestine (ulcerative chronic colitis of the rectum) using IL-1-β (3A6) Mouse mAb.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 24 Expand Image
    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.
    Mouse Reactive Senescence Associated Secretory Phenotype (SASP) Antibody Sampler Kit: Image 25 Expand Image
    Immunohistochemical analysis of paraffin embedded THP-1 cell pellets, control (left) or LPS-treated (right) using IL-1-β (3A6) Mouse mAb.
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    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.