Human Caspase-9 Antibody
R&D Systems, part of Bio-Techne | Catalog # AF8301
Key Product Details
Species Reactivity
Validated:
Human
Cited:
Human
Applications
Validated:
Western Blot
Cited:
Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
Product Specifications
Immunogen
E. coli-derived recombinant human Caspase-9
Val139-Asp330
Accession # P55211
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.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Scientific Data Images for Human Caspase-9 Antibody
Detection of Human Caspase-9 by Western Blot.
Western blot shows lysates of HEK293T human embryonic kidney cell line and HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Caspase-9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8301) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Caspase-9 at approximately 46 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 2.
Detection of Human Caspase-9 by Western Blot.
Western blot shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 ug/ml Staurosporine (STS) for 2 hours. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Caspase-9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8301) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for Caspase-9 at approximately 46, 37, 35 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 2.Applications for Human Caspase-9 Antibody
Application
Recommended Usage
Western Blot
1 µg/mL
Sample: Staurosporine-treated Jurkat human acute T cell leukemia cell line, HEK293T human embryonic kidney cell line, HepG2 human hepatocellular carcinoma cell line
Sample: Staurosporine-treated Jurkat human acute T cell leukemia cell line, HEK293T human embryonic kidney cell line, HepG2 human hepatocellular carcinoma cell line
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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-9
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.
Additional Caspase-9 Products
Product Documents for Human Caspase-9 Antibody
Product Specific Notices for Human Caspase-9 Antibody
For research use only
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