Human Caspase-3 Antibody Summary
aa 29-175
Accession # P42574
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of Human Precursor Caspase‑3 and p18 Subunit by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 µM staurosporine (STS) for 3 hours. PVDF membrane was probed with 1 µg/mL of Human Caspase-3 Monoclonal Antibody (Catalog # MAB707), followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). Specific bands were detected for precursor Caspase-3 and the p18 subunit at approximately 36 and 18 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.
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Western Blot Shows Human Caspase‑3 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Caspase-3 knockout HeLa cell line (KO). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Caspase-3 Monoclonal Antibody (Catalog # MAB707) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Caspase-3 at approximately 32 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Caspase-3
Caspase-3 (Cysteine-aspartic acid protease 3/Casp3; also Yama, apopain and CPP32) is a 29 kDa heterodimer that belongs to the peptidase C14A family of enzymes. It is widely expressed, and considered to be the major executioner caspase in the apoptotic cascade. Human procaspase-3 is a 32 kDa, 277 amino acid (aa) protein and is normally an inactive homodimer. Following cell stress/activation, procaspase-3 undergoes proteolysis to generate an N-terminal 148 aa p17/17 kDa subunit (aa 29-175), plus a 102 aa C-terminal p12/12 kDa subunit. These subunits noncovalently heterodimerize, and associate with another p17/p12 heterodimer to form an active enzyme. There is one potential variant that shows an alternative start site nine aa upstream of the standard start site coupled with a 21 aa substitution for aa 162-277. Over aa 29-175, human and mouse caspase-3 share 87% aa identity.
Product Datasheets
Citations for Human Caspase-3 Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 7
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Alpha1-antitrypsin protects lung cancer cells from staurosporine-induced apoptosis: the role of bacterial lipopolysaccharide
Authors: N Schwarz, S Tumpara, S Wrenger, E Ercetin, J Hamacher, T Welte, S Janciauski
Sci Rep, 2020-06-12;10(1):9563.
Species: Human
Sample Types: Cell Culture Supernates
Applications: Western Blot -
Preeclampsia is associated with alterations in the p53-pathway in villous trophoblast.
Authors: Sharp A, Heazell A, Baczyk D, Dunk C, Lacey H, Jones C, Perkins J, Kingdom J, Baker P, Crocker I
PLoS ONE, 2014-01-30;9(1):e87621.
Species: Human
Sample Types: Tissue Homogenates
Applications: Western Blot -
PMA synergistically enhances apicularen A-induced cytotoxicity by disrupting microtubule networks in HeLa cells.
Authors: Seo K, Kim J, Park J, Song K, Yun E, Park J, Kweon G, Yoon W, Lim K, Hwang B
BMC Cancer, 2014-01-22;14(0):36.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Interactions between important regulatory proteins and human alphaB crystallin.
Authors: Ghosh JG, Shenoy AK, Clark JI
Biochemistry, 2007-05-08;46(21):6308-17.
Species: Human
Sample Types: Recombinant Protein
Applications: ELISA-Based Protein Pin Array -
Developmental differences in the responses of IL-6 and IL-13 transgenic mice exposed to hyperoxia.
Authors: Choo-Wing R, Nedrelow JH, Homer RJ, Elias JA, Bhandari V
Am. J. Physiol. Lung Cell Mol. Physiol., 2007-03-30;293(1):L142-50.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC-P -
Proteolytically processed soluble tumor endothelial marker (TEM) 5 mediates endothelial cell survival during angiogenesis by linking integrin alpha(v)beta3 to glycosaminoglycans.
Authors: Vallon M, Essler M
J. Biol. Chem., 2006-09-17;281(45):34179-88.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Inhibition of lymphotoxin-beta receptor-mediated cell death by survivin-DeltaEx3.
Authors: You RI, Chou YC
Cancer Res., 2006-03-15;66(6):3051-61.
Species: Human
Sample Types: Cell Lysates
Applications: Immunoprecipitation
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The antibody works fine for what it is, but I think it will not work for our experiments. Compared to Jurkat or other cell types you only get very faint bands for caspase in RBCs unless you fix your membrane after the transfer, this gave a false positive comparing a negative and positive control for caspase activation. We had to switch to measuring samples only with flow. If your experiments are compatible it's a good antibody though. We tried this experiment with several antibodies of different companies and this one did give a nicer looking band.
