Rabbit Anti-Caspase-9 antibody

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Caspase-9 subunit p35; Apaf-3; APAF 3; APAF3; Apoptosis related cysteine peptidase; Apoptotic protease activating factor 3; Apoptotic protease MCH 6; Apoptotic protease MCH6; CASP 9; CASP9; Caspase 9; Caspase 9 apoptosis related cysteine protease; Caspase

Cat:

SL20773R

Species Reactivity：

Human,Mouse,Rat,(predicted: Pig,Cow,Rabbit,Sheep,)

Immunogen：

KLH conjugated synthetic peptide derived from human Caspase-9:231-330/416

Format：

Liquid

Storage instructions：

Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Concentration：

1mg/ml

Clonality：

Polyclonal

Isotype：

IgG

Applications：

WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500IHC-F=1:100-500Flow-Cyt=1ug/TestICC=1:100IF=1:100-500（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.

Host：

Rabbit

Product Overview：

Sample: K562 Cell (Human) Lysate at 40 ugPrimary: Anti-Caspase-9 (SL20773R) at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 35/50 kDObserved band size: 35 kDSample: Urinary bladder(Mouse) Lysate at 40 ugJurkat(Human) Cell Lysate at 30 ugPrimary: Anti-Caspase-9 (SL20773R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 46-51'35'37 kDObserved band size: 35 kDSample: Spleen (Mouse) Lysate at 40 ugPrimary: Anti-Caspase-9 (SL20773R) at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 35/50 kDObserved band size: 35 kDParaformaldehyde-fixed, paraffin embedded (rat pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Caspase-9) Polyclonal Antibody, Unconjugated (SL20773R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (mouse pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Caspase-9) Polyclonal Antibody, Unconjugated (SL20773R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (Rat urinary bladder); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Caspase-9) Polyclonal Antibody, Unconjugated (SL20773R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (Rat heart); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Caspase-9) Polyclonal Antibody, Unconjugated (SL20773R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.HepG2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, SLC0005) at 37°C for 20 min; Antibody incubation with (Caspase-9) polyclonal Antibody, Unconjugated (SL20773R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.Overlay histogram showing Hela cells(1ug/1x10^6 cells) stained with SL20773R (Green line).The cells were fixed with 80% methanol (5 min) and and then permeabilized with 0.01M PBS-Tween for 20 min . The cells were then incubated in 1x PBS/10% normal goat serum to block non-specific protein-protein interactions followed by the antibody for 30 min at 22℃. The secondary antibody used was fluorescein isothiocyanate goat anti-rabbit IgG (H+L) (SL0295G-FITC , Brillant blue line) at 1/200 dilution for 30 min at 22℃. Isotype control antibody was rabbit IgG (polyclonal,SL0295P,Orange line) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of 20,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

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Unit:

Price: $228.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
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Caspase 9 (also known as ICE like apoptotic protease 6 (ICE LAP6), apoptotic protease Mch6, and apoptotic protease activating factor 3 (Apaf3)) is a member of the peptidase family C14 that contains a CARD domain. This caspase is active as a heterotetramer and has been reported to have two isoforms. ProCaspase 9 has been reported to be approximately 47 kD. This caspase is present in the cytosol and, upon activation, translocates to the mitochondria. Caspase 9 is involved in the caspase activation cascade responsible for apoptosis execution and cleaves/activates Caspase 3 and Caspase 6. Caspase 9 is inhibited by the dominant negative isoform, BclXL, cIAP1, cIAP2, XIAP, and Livin. This caspase becomes activated when recruited to Apaf1/cytochrome c complex, and following cleavage by Apaf1, granzyme B, Caspase 3, possibly Caspase 8 and Caspase 10 into large p37 and small p10 subunits. Caspase 9 intereacts with BIRC7 and has been shown to cleave PARP and vimentin.

Function:
Involved in the activation cascade of caspases responsible for apoptosis execution. Binding of caspase-9 to Apaf-1 leads to activation of the protease which then cleaves and activates caspase-3. Proteolytically cleaves poly(ADP-ribose) polymerase (PARP).
Isoform 2 lacks activity is an dominant-negative inhibitor of caspase-9.

Subunit:
Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 35 kDa (p35) and a 10 kDa (p10) subunit. Caspase-9 and APAF1 bind to each other via their respective NH2-terminal CED-3 homologous domains in the presence of cytochrome C and ATP. Interacts (inactive form) with EFHD2. Interacts with HAX1. Interacts with BIRC2/c-IAP1, XIAP/BIRC4, BIRC5/survivin, BIRC6/bruce and BIRC7/livin.

Tissue Specificity:
Ubiquitous, with highest expression in the heart, moderate expression in liver, skeletal muscle, and pancreas. Low levels in all other tissues. Within the heart, specifically expressed in myocytes.

Post-translational modifications:
Cleavages at Asp-315 by granzyme B and at Asp-330 by caspase-3 generate the two active subunits. Caspase-8 and -10 can also be involved in these processing events.
Phosphorylated at Thr-125 by MAPK1/ERK2. Phosphorylation at Thr-125 is sufficient to block caspase-9 processing and subsequent caspase-3 activation.

Similarity:
Belongs to the peptidase C14A family.
Contains 1 CARD domain.

SWISS:
P55211

Gene ID:
842

Database links:

Entrez Gene: 842 Human

Entrez Gene: 12371 Mouse

Entrez Gene: 58918 Rat

Omim: 602234 Human

SwissProt: P55211 Human

SwissProt: Q4FJK5 Mouse

SwissProt: Q920G4 Rat

Unigene: 329502 Human

Unigene: 88829 Mouse

Unigene: 32199 Rat

Caspase-9半胱氨酸蛋白酶家族成员之一，又称ICE-Lap6（ICE Like apoptotease 6）参与细胞凋亡过程和细胞因子的加工过程，在许多胚胎和成人组织中都有分布。此抗体主要用于肿瘤研究。 Picture

Sample:
Urinary bladder(Mouse) Lysate at 40 ug
Jurkat(Human) Cell Lysate at 30 ug
Primary: Anti-Caspase-9 (SL20773R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 46-51'35'37 kD
Observed band size: 35 kD

Sample:
Spleen (Mouse) Lysate at 40 ug
Primary: Anti-Caspase-9 (SL20773R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 35/50 kD
Observed band size: 35 kD

Paraformaldehyde-fixed, paraffin embedded (rat pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Caspase-9) Polyclonal Antibody, Unconjugated (SL20773R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Caspase-9) Polyclonal Antibody, Unconjugated (SL20773R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Rat urinary bladder); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Caspase-9) Polyclonal Antibody, Unconjugated (SL20773R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Rat heart); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Caspase-9) Polyclonal Antibody, Unconjugated (SL20773R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

HepG2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, SLC0005) at 37°C for 20 min; Antibody incubation with (Caspase-9) polyclonal Antibody, Unconjugated (SL20773R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Overlay histogram showing Hela cells(1ug/1x10^6 cells) stained with SL20773R (Green line). The cells were fixed with 80% methanol (5 min) and and then permeabilized with 0.01M PBS-Tween for 20 min . The cells were then incubated in 1x PBS/10% normal goat serum to block non-specific protein-protein interactions followed by the antibody for 30 min at 22℃. The secondary antibody used was fluorescein isothiocyanate goat anti-rabbit IgG (H+L) (SL0295G-FITC , Brillant blue line) at 1/200 dilution for 30 min at 22℃. Isotype control antibody was rabbit IgG (polyclonal,SL0295P,Orange line) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of 20,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

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