Human/Mouse Caspase-3 Antibody

(5 citations)   
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human Caspase-3.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human Caspase-3
    Met1-His277
    Accession # AAA65015
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.5 µg/mL
    See below
  • Simple Western
    5 µg/mL
    See below
  • Immunoprecipitation
    1 µg/106 cells
    See below
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.
Data Examples

Detection of Human and Mouse Caspase‑3 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line and DA3 mouse myeloma cell line untreated (-) or treated (+) with 1 µg/mL Staurosporine (STS) for 12 hours. PVDF Membrane was probed with 0.5 µg/mL of Goat Anti-Human Caspase‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-605-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for Caspase‑3 precursor at approximately 34 kDa (as indicated) and Caspase‑3 (cleaved) at approximately 18 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Immunoprecipitation

Immunoprecipitation of Human Caspase‑3. Jurkat human acute T cell leukemia cell line was treated with indicated concentrations of Staurosporine for 0 or 4 hours. Caspase‑3 was immunoprecipitated from lysates of 1‑2 x 106 cells following incubation with 1 or 4 µg Goat Anti-Human Caspase‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-605-NA) overnight at 4 ºC. Caspase‑3-antibody complexes were absorbed using Protein G expressing Staph cells (Sigma). Immunoprecipitated Caspase‑3 was detected by Western blot using 0.5 µg/mL Goat Anti-Human Caspase‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF‑605‑NA). View our recommended buffer recipes for immunoprecipitation.

Detection of Human Caspase‑3 by Simple WesternTM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, HepG2 human hepatocellular carcinoma cell line, and Jurkat human acute T cell leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for Caspase‑3 at approximately 40 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse Caspase‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-605-NA) 1:50 dilution followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 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 member of the peptidase C14A family of enzymes (1, 2, 3). It is widely expressed and is an integral component of the apoptotic cascade. Caspase-3 is considered to be the major executioner caspase; that is, the primary downstream mediator of apoptotic-associated proteolysis (2, 3, 4). Active Caspase-3 is known to utilize a Cys residue to cleave multiple substrates, including PARP, proIL-16, PKC-gamma & -δ, procaspases 6, 7 and 9, and beta -catenin (1). Human procaspase-3 is a 32 kDa, 277 amino acid (aa) protein (5, 6, 7). Normally, it is an inactive, cytosolic homodimer, but following an upstream signal that activates processing proteases, procaspase-3 undergoes proteolytic cleavage (1, 2, 8, 9). This generates an N-terminal 175 aa p20/20 kDa subunit plus a 102 aa C-terminal p12/12 kDa subunit, followed by further processing of the p20 subunit at Asp28 to generate a final p17 subunit (aa 29 - 175) (9). The p17 and p12 subunits noncovalently heterodimerize, and subsequently associate with another p17/p12 heterodimer to form an active antiparallel homodimer. The p17 subunit contains the enzyme active site (aa 161 - 165), with an embedded catalytic Cys which is normally nitrosylated and inactive. Full activation requires both proteolytic processing and Cys163 denitrosylation (10). Multiple proteases can use Caspase-3 as a substrate including Caspase-6, -8, and -10, granzyme B, and Caspase-3 itself (9, 11, 12, 13). 
  • References:
    1. Chowdhury, I. et al. (2008) Comp. Biochem. Physiol. B 151:10.
    2. Boatright, K.M. & G.S. Salvesen (2003) Curr. Opin. Cell Biol. 15:725.
    3. Launay, S. et al. (2005) Oncogene 24:5137.
    4. Walsh, J.G. et al. (2008) Proc. Natl. Scad. Sci. USA 105:12815.
    5. Nicholson, D.W. et al. (1995) Nature 376:37.
    6. Tewari, M. et al. (1995) Cell 81:801.
    7. Fernandes-Alnemri, T. et al. (1994) J. Biol. Chem. 269:30761.
    8. Milisav, I. et al. (2009) Apoptosis 14:1070.
    9. Han, Z. et al. (1997) J. Biol. Chem. 272:13432.
    10. Rossig, L. et al. (1999) J. Biol. Chem. 274:6823.
    11. Rank, K.B. et al. (2001) Protein Expr. Purif. 22:258.
    12. Atkinson, E.A. et al. (1998) J. Biol. Chem. 273:21261.
    13. Cohen, G.M. (1997) Biochem. J. 326:1.
  • Entrez Gene IDs:
    836 (Human); 12367 (Mouse)
  • Alternate Names:
    Apopain; apoptosis-related cysteine protease; CASP3; CASP-3; caspase 3, apoptosis-related cysteine peptidase; Caspase3; Caspase-3; CPP32; CPP-32; CPP32B; CPP32SREBP cleavage activity 1; Cysteine protease CPP32; EC 3.4.22; EC 3.4.22.56; LICE-1; PARP cleavage protease; procaspase3; Protein Yama; SCA-1; YAMA
Related Research Areas
Citations:

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.

5 Citations: Showing 1 - 5
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Species
Applications
Sample Type
  1. Bok is a genuine multi-BH-domain protein that triggers apoptosis in the absence of Bax and Bak and augments drug response
    Authors: S Einsele-Sc, S Malmsheime, K Bertram, D Stehle, J Johänning, M Manz, PT Daniel, BF Gillissen, K Schulze-Os, F Essmann
    J Cell Sci, 2016;0(0):.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. Overexpressing cellular repressor of E1A-stimulated genes protects mesenchymal stem cells against hypoxia- and serum deprivation-induced apoptosis by activation of PI3K/Akt.
    Authors: Deng J, Han Y, Yan C, Tian X, Tao J, Kang J, Li S
    Apoptosis, 2010;15(4):463-73.
    Species: Rat
    Sample Type: Cell Lysates
    Application: WB
  3. Ellipticine derivative NSC 338258 represents a potential new antineoplastic agent for the treatment of multiple myeloma.
    Authors: Tian E, Landowski TH, Stephens OW, Yaccoby S, Barlogie B, Shaughnessy JD
    Mol. Cancer Ther., 2008;7(3):500-9.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  4. Photochemically mediated delivery of AdhCMV-TRAIL augments the TRAIL-induced apoptosis in colorectal cancer cell lines.
    Authors: Engesaeter BØ, Bonsted A, Lillehammer T, Engebraaten O, Berg K, Maelandsmo GM
    Cancer Biol. Ther., 2006;5(11):1511-20.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  5. The B30.2 domain of pyrin, the familial Mediterranean fever protein, interacts directly with caspase-1 to modulate IL-1beta production.
    Authors: Chae JJ, Wood G, Masters SL, Richard K, Park G, Smith BJ, Kastner DL
    Proc. Natl. Acad. Sci. U.S.A., 2006;103(26):9982-7.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
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