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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.
Figure 1 Independent Antibody Validation (IAV) via Protein Expression Profile in Human Cell Lines
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.
IAV
Trial Size

Caspase-9 Antibody

CATALOG NUMBER: 2073

Clonality:
Polyclonal
Tested Applications:
ELISA, ICC, IF, IP, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
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.
Conjugate:
Unconjugated
Tested Applications:
ELISA, ICC, IF, IP, WB
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.
Specificity:
Caspase-9 antibody is predicted to have no cross reactivity to other members in the caspase family.
Positive Control 1:
Cat. No. 1201 - HeLa Cell Lysate
Positive Control 2:
Cat. No. 17-004 - K-562 Cell Slide
Predicted Molecular Weight:
Predicted: 46kD
Observed: 46 kD
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.

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.
Purification:
Caspase-9 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
Caspase-9 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
Caspase-9 antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
CASP9
Additional Names:
Caspase-9 Antibody: MCH6, APAF3, APAF-3, PPP1R56, ICE-LAP6, MCH6, Caspase-9, Apoptotic protease Mch-6, CASP-9
Protein Accession Number:
P55211
Protein Gi Number:
28558771
Ncbi Gene Id Number:
842
User Note:
Optimal dilutions for each application to be determined by the researcher.
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 Reference 1:
Duan et al. J Biol Chem 1996;271:16720-4.
Background Reference 2:
Srinivasula et al. J Biol Chem 1996;271:27099-106.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

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.

CATALOG NUMBER:

2073

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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