Phospho-Caspase 9 (Ser196) Recombinant antibody

Phospho-Caspase 9 (Ser196) Uni-rAb^TM Recombinant Antibody for WB, IF/ICC, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

WB, IF/ICC, ELISA

Conjugate

Unconjugated

CloneNo.

3P16

Cat no : 80346-1-RR

Synonyms

CASP9, Caspase 9, Caspase 9/p35/p10, Apoptotic protease Mch-6, Apoptotic protease Mch 6

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Validation Data Gallery

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  WB analysis using 80346-1-RR

  Non-treated HEK-293 and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 80346-1-RR (Phospho-Caspase 9 (Ser196) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Tubulin antibody as loading control.

• 

  WB analysis using 80346-1-RR

  Non-treated Jurkat and Calyculin A treated Jurkat cells were subjected to SDS PAGE followed by western blot with 80346-1-RR (Phospho-Caspase 9 (Ser196) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Tubulin antibody as loading control.

• 

  WB analysis using 80346-1-RR

  Non-treated NIH/3T3 and Calyculin A treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 80346-1-RR (Phospho-Caspase 9 (Ser196) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Tubulin antibody as loading control.

• 

  IF Staining of HeLa using 80346-1-RR

  Immunofluorescent analysis of (4% PFA) fixed Calyculin A treated HeLa cells using Phospho-Caspase 9 (Ser196) antibody (80346-1-RR, Clone: 3P16 ) at dilution of 1:3075 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002).

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Tested Applications

• Positive WB detected in: NIH/3T3 cells, HEK-293 cells, Jurkat cells, Calyculin A treated HEK-293 cells, Calyculin A treated Jurkat cells, Calyculin A treated NIH/3T3 cells
• Positive IF/ICC detected in: Calyculin A treated HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:1537-1:6150

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

80346-1-RR targets Phospho-Caspase 9 (Ser196) in WB, IF/ICC, ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: Peptide
• Full Name: caspase 9, apoptosis-related cysteine peptidase
• Calculated Molecular Weight: 46 kDa
• Observed Molecular Weight: 46 kDa, 35 kDa
• GenBank Accession Number: BC002452
• Gene Symbol: Caspase 9
• Gene ID (NCBI): 842
• RRID: AB_2918890
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Caspase 9 also name as MCH6, APAF3, APAF-3, ICE-LAP6 and CASPASE-9c, is a member of the cysteine-aspartic acid protease (caspase) family. It's synthesized as a 46kDa precursor protein which can be cleaved into a 35kDa subunit and a 11kDa subunit. The phosphorylated type can be detected at 55kDa and 35kDa. It plays a central role in the mitochondrial or intrinsic apoptotic pathway that is engaged in response to many apoptotic stimuli. Once activated, caspase-9 cleaves and activates the effector caspases 3 and 7 to bring about apoptosis. It can be phosphorylated by PKB/AKT1 at Ser196, this modification will downregulate its activity and decrease apoptosis. Akt phosphorylation site found in human caspase 9 is absent in mouse caspase 9.It's reported that there is an increase in caspase 9 expression and activity in the hypoxic brain. Inhibition of Caspase 9 activity would render opportunity to treat neurological diseases such as stroke, neurodegenerative diseases or brain injury caused by hypoxia. (PMID: 19788417, PMID: 10529400, PMID: 9812896, PMID: 18840507) In recent years, the localization of caspase9 was a focus of interest. Beside its cytoplasmic distribution, a very extensive localization study was done on rat brain tissue, where caspase9 was found located predominantly in the nucleus and to a lesser extend in the cytoplasm [PMID: 15541731].