ProSci

Caspase-9 Antibody

Product Code:
 
PSI-2073
Product Group:
 
Primary Antibodies
Supplier:
 
ProSci
Host Type:
 
Rabbit
Antibody Isotype:
 
IgG
Antibody Clonality:
 
Polyclonal
Regulatory Status:
 
RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunofluorescence (IF)
  • Immunohistochemistry (IHC)
  • Western Blot (WB)
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<strong>Figure 1 Independent Antibody Validation (IAV) via Protein Expression Profile in Human Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: Caspase 9, 2071 (1 μg/mL), Caspase 9, 2073 (1 μg/mL) and beta-actin (1.5 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 2 Western Blot Validation in Human Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: : Caspase 9, 2073 (1 μg/mL)), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 3 Western Blot Validation in Mouse Cell Line</strong><br> Loading: 15 μg of 3T3/NIH cell lysate. Antibodies: : Caspase 9, 2073 (1 μg/mL)), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 4 Immunofluorescence Validation of Caspase 9 in K562 Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling Caspase 9 with 2073 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green).
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<strong>Figure 5 Immunofluorescence Validation of Caspase 9 in HeLa Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa cells labeling Caspase 9 with 2073 at 5 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
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<strong>Figure 6 Immunocytochemistry Validation of Caspase 9 in HeLa Cells</strong><br> Immunocytochemical analysis of HeLa cells using anti-Caspase 9 antibody (2073) at 5 μg/ml. Cells was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
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<strong>Figure 1 Independent Antibody Validation (IAV) via Protein Expression Profile in Human Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: Caspase 9, 2071 (1 μg/mL), Caspase 9, 2073 (1 μg/mL) and beta-actin (1.5 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 Western Blot Validation in Human Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: : Caspase 9, 2073 (1 μg/mL)), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Western Blot Validation in Mouse Cell Line</strong><br> Loading: 15 μg of 3T3/NIH cell lysate. Antibodies: : Caspase 9, 2073 (1 μg/mL)), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Immunofluorescence Validation of Caspase 9 in K562 Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling Caspase 9 with 2073 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green).
<strong>Figure 5 Immunofluorescence Validation of Caspase 9 in HeLa Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa cells labeling Caspase 9 with 2073 at 5 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
<strong>Figure 6 Immunocytochemistry Validation of Caspase 9 in HeLa Cells</strong><br> Immunocytochemical analysis of HeLa cells using anti-Caspase 9 antibody (2073) at 5 μg/ml. Cells was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

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PSI-2073-0.02mg0.02mg£150.00
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Further Information

Additional Names:
Caspase-9 Antibody: MCH6, APAF3, APAF-3, PPP1R56, ICE-LAP6, MCH6, Caspase-9, Apoptotic protease Mch-6, CASP-9
Application Note:
WB: 1 μg/mL; ICC: 2- 5 μg/mL; IF: 5-20 μg/mL.

Antibody validated: Western Blot in human and mouse samples, Immunocytochemistry, Immunofluorescence in human samples. All other applications and species not yet tested.
Background:
Caspase-9 Antibody: Apoptosis is related to many diseases and induced by a family of cell death receptors and their ligands. Cell death signals are transduced by death domain containing adapter molecules and members of the caspase family of proteases. A novel member in the caspase family was recently identified and designated ICE-LAP6, Mch6, and Apaf-3. Caspase-9 and Apaf-1 bind to each other, which leads to caspase-9 activation. Caspase-9 is also activated by granzyme B and CPP32. Activated caspase-9 cleaves and activates caspase-3 that is one of the key proteases, being responsible for the proteolytic cleavage of many key proteins in apoptosis. Caspase-9 play a central role in cell death induced by a wide variety of apoptosis activators including TNFα, TRAIL, anti-CD-95, FADD, and TRADD. Caspase-9 is expressed in a variety of human tissues.
Background References:
  • Duan et al. J Biol Chem 1996;271:16720-4.
  • Srinivasula et al. J Biol Chem 1996;271:27099-106.
Buffer:
Caspase-9 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Conjugate:
Unconjugated
DISCLAIMER:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.
Homology:
Predicted species reactivity based on immunogen sequence: Rat (50%).
Immunogen:
Anti-Caspase 9 antibody (2073) was raised against a peptide corresponding to 20 amino acids near the center of human Caspase 9.

The immunogen is located within amino acids 290-340 of Caspase 9.
ISOFORMS:
Human CASPASE 9 has 4 isoforms, including isoform 1 (416aa, 46kD), isoform 2 (266aa, 30kD), isoform 3 (154aa, 17kD), and isoform 4 (333aa, 37kD). Mouse CASPASE 9 has one isoform (454aa, 50kD) and Rat CASPASE 9 also has one isoform (454aa, 50kD). 2073 can detect human and mouse isoform.
NCBI Gene ID #:
842
NCBI Official Name:
caspase 9, apoptosis-related cysteine peptidase
NCBI Official Symbol:
CASP9
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 46kD

Observed: 46 kD
Protein Accession #:
P55211
Protein GI Number:
28558771
Purification:
Caspase-9 Antibody is affinity chromatography purified via peptide column.
Research Area:
Apoptosis,Cancer
SPECIFICITY:
Caspase-9 antibody is predicted to have no cross reactivity to other members in the caspase family.
Swissprot #:
P55211
User NOte:
Optimal dilutions for each application to be determined by the researcher.
VALIDATION:

Independent Antibody Validation in Cell lines (Figure 1) shows similar CASPASE 9 expression profile in human cell lines detected by two independent anti-CASPASE 9 antibodies that recognize different epitopes, 2071 against N-terminus domain and 2073 against central domain.  CASPASE 9 proteins are detected in the most tested cell lines at different expression levels by the two independent antibodies.