Rabbit Anti-caspase-9 p10 antibody

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APAF 3; APAF3; Apoptosis related cysteine peptidase; Apoptotic protease activating factor 3; Apoptotic protease MCH 6; Apoptotic protease MCH6 antibody CASP 9; CASP9; Caspase 9; Caspase 9 apoptosis related cysteine protease; Caspase 9 precursor; Caspase 9

Cat:

SL8502R

Species Reactivity：

Human,Mouse,Rat,(predicted: Chicken,Dog,Pig,Cow,Horse,Rabbit,)

Immunogen：

KLH conjugated synthetic peptide derived from human caspase-9 subunit p10:351-416/416

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=1μg /TestIF=1:50-200（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: Brain (Mouse) Lysate at 40 ugLung (Mouse) Lysate at 40 ugPrimary: Anti-caspase-9 p10 (SL8502R) at 1/300 dilutionSecondary: HRP conjugated Goat-Anti-rabbit IgG (SL0295G-HRP) at 1/5000 dilutionPredicted band size: 10/50 kDObserved band size: 50 kDSample: Lane 1: Esophagus (Mouse) Lysate at 40 ugLane 2: Stomach (Mouse) Lysate at 40 ugLane 3: Urinary bladder (Mouse) Lysate at 40 ugPrimary: Anti-caspase-9 p10 (SL8502R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 46-51/37/35/10 kDObserved band size: 46 kDSample: Jurkat(Human) Cell Lysate at 30 ugHela(Human) Cell Lysate at 30 ugHepG2(Human) Cell Lysate at 30 ugPrimary: Anti-caspase-9 p10 (SL8502R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 46-51/37/35/10 kDObserved band size: 46/37 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 p10) Polyclonal Antibody, Unconjugated (SL8502R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti-caspase-9 p10 Polyclonal Antibody, Unconjugated(SL8502R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) stainingBlank control:K562 (fixed with 80% methanol (5 min) and and then permeabilized with 0.01M PBS-Tween for 20 min). Primary Antibody:Rabbit Anti-caspase-9 p10 antibody (SL8502R,Green); Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA; Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions; Secondary Antibody: Goat anti-rabbit IgG-FITC(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.Blank control:K562. Primary Antibody (green line): Rabbit Anti-caspase-9 p10 antibody (SL8502R) Dilution: 2μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-FITCDilution: 0.5μg /test. ProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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Unit:

Price: $228.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
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A unique family of cysteine proteases has been described that differs in sequence, structure and substrate specificity from any previously described protease family. This family, Ced-3/caspase-1, is comprised of caspase-1, caspase-2, caspase-3, caspase-4, caspase-6, caspase-7 (also designated Mch3, ICE-LAP3 or CMH-1), caspase-9 and caspase-10. Ced-3/caspase-1 family members function as key components of the apoptotic machinery and act to destroy specific target proteins which are critical to cellular longevity. Poly(ADP-ribose) polymerase plays an integral role in surveying for DNA mutations and double strand breaks. Caspase-3, caspase-7 and caspase-9, but not caspase-1, have been shown to cleave the nuclear protein PARP into an apoptotic fragment. Caspase-6, but not caspase-3, has been shown to cleave the nuclear lamins which are critical to maintaining the integrity of the nuclear envelope and cellular morphology. Caspase-10 has been shown to activate caspase-3 and caspase-7 in response to apoptotic stimuli.

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

Picture

Sample:
Lane 1: Esophagus (Mouse) Lysate at 40 ug
Lane 2: Stomach (Mouse) Lysate at 40 ug
Lane 3: Urinary bladder (Mouse) Lysate at 40 ug
Primary:
Anti-caspase-9 p10 (SL8502R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 46-51/37/35/10 kD
Observed band size: 46 kD

Sample:
Jurkat(Human) Cell Lysate at 30 ug
Hela(Human) Cell Lysate at 30 ug
HepG2(Human) Cell Lysate at 30 ug
Primary: Anti-caspase-9 p10 (SL8502R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 46-51/37/35/10 kD
Observed band size: 46/37 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 p10) Polyclonal Antibody, Unconjugated (SL8502R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-caspase-9 p10 Polyclonal Antibody, Unconjugated(SL8502R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining

Blank control:K562 (fixed with 80% methanol (5 min) and and then permeabilized with 0.01M PBS-Tween for 20 min).
Primary Antibody:Rabbit Anti-caspase-9 p10 antibody (SL8502R,Green); Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions;
Secondary Antibody: Goat anti-rabbit IgG-FITC(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.

Blank control:K562.
Primary Antibody (green line): Rabbit Anti-caspase-9 p10 antibody (SL8502R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 0.5μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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