Caspase-9 Antibody

Western blot shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 μg/mL staurosporine (STS) for for the indicated times. PVDF membrane was probed with 1 µg/mL of Human Caspase‑9 Polyclonal Antibody (Catalog # AF8301), followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). Specific bands were detected for Caspase‑9 precursor and pro + LSU at approximately 46 and 37 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 6.

• Reactivity: Hu
• Applications: WB, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Caspase-9 Antibody Summary

• Immunogen: E. coli-derived recombinant human Caspase-9
  Val139-Asp330
  Accession # P55211
• Specificity: Detects human Caspase-9 precursor and the pro + large subunit (LSU) that migrates as a 37 kDa band on SDS-PAGE generated by cleavage at Asp330. Does not detect the pro + LSU that migrates as a 34 kDa band on SDS-PAGE generated by cleavage at Asp315. Therefore, the major epitope detected occurs between Asp315 and Asp330.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CASP9
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/mL
  • Immunocytochemistry 5-15 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Purity: Immunogen affinity purified
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Caspase-9 Antibody

• APAF-3APAF3
• apoptotic protease activating factor 3
• Apoptotic protease Mch-6
• Apoptotic protease-activating factor 3
• CASP9
• CASP-9
• caspase 9, apoptosis-related cysteine peptidase
• Caspase9
• Caspase-9
• EC 3.4.22.62
• ICE-LAP6CASPASE-9c
• ICE-like apoptotic protease 6
• MCH6apoptosis-related cysteine protease

Background

Caspase-9 (Cysteine-aspartic acid protease 9/Casp-9; also APAF-3, Mch6 and ICE-LAP6) is a 35-37 kDa member of the peptidase C14A family of enzymes. Casp-9 is an initiator caspase that is part of the intrinsic apoptosis pathway. It is widely expressed and is particularly important during development. Human proCaspase-9 is a 47-48 kDa, 416 amino acid (aa) protein and it contains one CARD region (aa 1-92) and catalytic residues at His237 and Cys287. Following mitochondrial disruption, cytochrome c is released from mitochrondria. Cytochrome c acts on APAF-1, which induces procaspase-9 dimerization. The act of dimerization activates proCasp-9, leading to either the activation of Casp-3, or the autocleavage of proCasp-9, generating a 35 kDa subunit (aa 1-315) and a 12 kDa subunit. Activated Casp-3 will also act on proCasp-9, generating a 37 kDa subunit (aa 1-330) and a 10 kDa subunit (aa 331-416). These subunits associate to form an active heterotetramer. Casp-9 has an alternative start site at Met84 and a deletion of aa 140-289 that generates a dominant negative, 31 kDa isoform. Over aa 1-134, human Casp-9 shares 81% aa identity with mouse Casp-9; over aa 139-330, human Casp-9 shares 73% aa identity with mouse Casp-9.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Caspase-9 Antibody (AF8301)(2)

Publications using AF8301

• Tian E, Landowski TH, Stephens OW, Yaccoby S, Barlogie B, Shaughnessy JD Ellipticine derivative NSC 338258 represents a potential new antineoplastic agent for the treatment of multiple myeloma. Mol. Cancer Ther., 2008;7(3):500-9. 2008 [PMID: 18319333] (Western Blot, Human)
• Chae JJ, Wood G, Masters SL, Richard K, Park G, Smith BJ, Kastner DL The B30.2 domain of pyrin, the familial Mediterranean fever protein, interacts directly with caspase-1 to modulate IL-1beta production. Proc. Natl. Acad. Sci. U.S.A., 2006;103(26):9982-7. 2006 [PMID: 16785446] (Western Blot, Human)

FAQs for Caspase-9 Antibody (AF8301). (Showing 1 - 1 of 1 FAQs).

1. Could I use HeLa cell lysate as a positive control for Caspase 9 primary antibodies in WB?
   • After looking through our testing it does appear you should be fine using HeLa as a positive control cell lysate for Caspase 9 in Western Blot, as we have used this sample successfully in our QC tests and shown expression of Caspase 9.

Bioinformatics

• Gene Symbol: CASP9
• Uniprot:
  • P55211()