The CellSimple™ Cellular Stress Antibody Assay Kit is designed for use with the CellSimple™ Cell Analyzer. The kits allows for the quick and easy measurement of the extent of cellular stress or DNA damage under given experimental conditions.
The conjugated antibodies in this kit are supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibodies. Protect from light. Do not freeze. The remaining components of the kit should be stored at the recommended temperature upon receipt. All components in this kit are stable for at least 12 months when stored at the recommended temperature and left unused.
Phospho-p38 MAPK (Thr180/Tyr182) (3D7) Rabbit mAb (PE conjugate) recognizes endogenous levels of p38 MAPK only when dually phosphorylated at
Thr180/Tyr182. This antibody does not cross-react with other MAPK family members. Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb (Alexa Fluor® 488 Conjugate) recognizes H2A.X only when phosphorylated at Ser139. This antibody does not cross react with other phosphorylated histone proteins.
The CellSimple™ Cell Analyzer is a benchtop instrument that utilizes a disposable thin-film cassette and a combination of a 488 nm laser, two photomultiplier tubes (525/45 nm and 561 nm LP filters), Coulter Principle-based cell measurements, and on-board software to provide easy-to-run applications and data analysis. Data acquisition occurs within approximately 10 seconds per test. The instrument relies on disposable cassettes for sample handling, which alleviates the need for flow cell cleaning and fluidics maintenance and the instrument is small enough to be portable between the lab bench and the hood. Applications include quantitative assessments of cell viability, apoptosis, other labeled antibody markers and single and multiplexed bead-based assays for protein and cellular analysis.
p38 MAP kinase (MAPK) is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharide (LPS), UV light, and growth factors. p38 MAPK is activated by phosphorylation at Thr180 and Tyr182 by various MAPK kinases. Once activated the p38 MAPK phosphorylates a wide spectrum of cellular targets to help the cells adapt and cope with noxious conditions (1). Histone H2A.X is required for checkpoint-mediated cell cycle arrest and DNA repair following double-stranded DNA breaks. DNA damage, caused by ionizing radiation, UV-light, or radiomimetic agents results in rapid phosphorylation of H2A.X at Ser139. This very early event in the DNA-damage response is required for recruitment of a multitude of DNA-damage response proteins. In addition to its role in DNA-damage repair, H2A.X is required for DNA fragmentation during apoptosis and is phosphorylated by various kinases in response to apoptotic signals (2).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. CellSimple is a trademark of Cell Signaling Technology, Inc. XP is a registered trademark of Cell Signaling Technology, Inc. Alexa Fluor is a registered trademark of Life Technologies Corporation. The Alexa Fluor dye conjugates in this product are sold under license from Life Technologies Corporation, for research use only excluding use in combination with DNA microarrays and high content screening (HCS). This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or firstname.lastname@example.org.