Rabbit Anti-Phospho-Caspase-9 (Thr125) antibody

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APAF 3; APAF3; Apoptosis related cysteine peptidase; Apoptotic protease activating factor 3; Apoptotic protease Mch 6; CASP 9; CASP9; Caspase 9 Dominant Negative; Caspase 9 precursor; Caspase 9c; Caspase9; EC 3.4.22.; ICE LAP6; ICE like apoptotic protease

Cat:

SL3082R

Species Reactivity：

Human,Mouse,Rat,

Immunogen：

KLH conjugated Synthesised phosphopeptide derived from human Caspase-9 around the phosphorylation site of Thr125:PE(p-T)PR

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=0.2μg /testIF=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: Heart (Rat) Lysate at 40 ugHeart (Mouse) Lysate at 40 ugPrimary: Anti-Phospho-Caspase-9 (Thr125) (SL3082R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 46-51/35/37 kDObserved band size: 51/37/35 kDSample: Brain(Rat)Lysate at 40 ugPrimary: Anti-p-Caspase-9 (Thr125)(SL3082R)at 1/300 dilutionSecondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilutionPredicted band size: 35kDObserved band size: 35kDSample:H9C2 (Rat)cell Lysate at 40 ugPrimary: Anti-p-Caspase-9 (Thr125)(SL3082R)at 1/300 dilutionSecondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilutionPredicted band size: 35kDObserved band size: 35kDParaformaldehyde-fixed, paraffin embedded (rat brain); 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 (P-caspase-9(Thr125)) Polyclonal Antibody, Unconjugated (SL3082R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining. Paraformaldehyde-fixed, paraffin embedded (rat brain); 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 (P-caspase-9(Thr125)) Polyclonal Antibody, Unconjugated (SL3082R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining. Paraformaldehyde-fixed, paraffin embedded (Mouse brain); 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 (Phospho-Caspase-9 (Thr125)) Polyclonal Antibody, Unconjugated (SL3082R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Blank control (blue line): Hela (fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min on ice). Primary Antibody (green line): Rabbit Anti-Phospho-Caspase-9 (Thr125) antibody (SL3082R), Dilution: 0.2μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1μg /test.

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Unit:

Price: $256.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
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This gene encodes a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. This protein can undergo autoproteolytic processing and activation by the apoptosome, a protein complex of cytochrome c and the apoptotic peptidase activating factor 1; this step is thought to be one of the earliest in the caspase activation cascade. This protein is thought to play a central role in apoptosis and to be a tumor suppressor. Alternative splicing results in multiple transcript variants. [provided by RefSeq, May 2013]

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 with the inhibitors BIRC2, BIRC4, BIRC5 and BIRC7. Interacts (inactive form) with EFHD2. Interacts with HAX1.

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凋亡的研究。 Picture

Sample: Brain(Rat)Lysate at 40 ug
Primary: Anti-p-Caspase-9 (Thr125)(SL3082R)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilution
Predicted band size: 35kD
Observed band size: 35kD

Sample:H9C2 (Rat)cell Lysate at 40 ug
Primary: Anti-p-Caspase-9 (Thr125)(SL3082R)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilution
Predicted band size: 35kD
Observed band size: 35kD

Paraformaldehyde-fixed, paraffin embedded (rat brain); 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 (P-caspase-9(Thr125)) Polyclonal Antibody, Unconjugated (SL3082R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Mouse brain); 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 (Phospho-Caspase-9 (Thr125)) Polyclonal Antibody, Unconjugated (SL3082R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Blank control (blue line): Hela (fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody (green line): Rabbit Anti-Phospho-Caspase-9 (Thr125) antibody (SL3082R), Dilution: 0.2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1μg /test.

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