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Home > Product > Antibody > Rabbit Anti-Caspase 3 precursor antibody
Caspase 3 precursor; APOPAIN; CASP3; Caspase 3 apoptosis related cysteine protease; Caspase3; CPP32; CPP32B; Cysteine protease CPP32; Human cysteine protease CPP32 isoform alpha mRNA complete cds; PARP cleavage protease; SCA 1; SCA1; SREBP cleavage activi
Cat:
SL2593R
Species Reactivity：
Human,Mouse,Rat,(predicted: Dog,Rabbit,)
Immunogen：
KLH conjugated synthetic peptide derived from human Caspase 3 precursor:11-120/277
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=1:100-500（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: Lane 1: Spleen (Mouse) Lysate at 40 ugLane 2: Lung (Mouse) Lysate at 40 ugPrimary: Anti-Caspase 3 precursor (SL2593R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 32 kDObserved band size: 32 kDSample: Liver (Mouse) Lysate at 40 ugPrimary: Anti-Caspase 3 precursor  (SL2593R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 28 kDObserved band size: 28 kDSample: Lane1: Brain (Mouse) Lysate at 30 ugLane2: Liver (Mouse) Lysate at 30 ugPrimary: Anti-pro-caspase-3 (SL2593R) at 1:300 dilution; Secondary: HRP conjugated Goat Anti-Rabbit IgG(SL0295G-HRP) at 1:3000 dilution; Predicted band size : 28kDObserved band size : 30kDTissue/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-pro-caspase-3 Polyclonal Antibody, Unconjugated(SL2593R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) stainingBlank control (blue line): Hela (fixed with 80% methanol (5 min at -20℃) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature). Primary Antibody (green line): Rabbit Anti-Caspase 3 precursor antibody (SL2593R), Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1μg /test. Blank control: NIH/3T3. Primary Antibody (green line): Rabbit Anti-Caspase 3 precursor antibody (SL2593R) Dilution: 2μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-AF488Dilution: 1μg /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.Blank control (blue line): HL60 (fixed with 2% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature). Primary Antibody (green line): Rabbit Anti-Caspase 3 precursor antibody (SL2593R), Dilution: 1μg /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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Product PDFs
Datasheet:


Caspases are a family of cysteine proteases that are key mediators of programmed cell death or apoptosis. The precursor form of all caspases is composed of a prodomain, and large and small catalytic subunits. The active forms of caspases are generated by several stimuli including ligand-receptor interactions, growth factor deprivation and inhibitors of cellular functions. All known caspases require cleavage adjacent to aspartates to liberate one large and one small subunit, which associate into a2b2 tetramer to form the active enzyme. Gene for Caspase 3 also known as Yama, CPP32, and apopain codes for a 32-kDa protein. Caspase 3 cleaves the death substrate poly(ADP-ribose) polymerase (PARP) to a specific 85 kDa form observed during apoptosis and is inhibitable by the CrmA protein. Other Caspase 3 substrates include DNA-PK, actin, GAS2, and procaspase-6, etc. Caspase 3 is activated by cleavage events at Asp-28/Ser-29 (between N-terminal pro-domain) and Asp-175/Ser-176 (between large and small subunits) to generate a large subunit of 17-kDa and a small subunit of 12-kDa.

Function:
Involved in the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp-|-Gly-217' bond. Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9. Involved in the cleavage of huntingtin. Triggers cell adhesion in sympathetic neurons through RET cleavage.

Subunit:
Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 17 kDa (p17) and a 12 kDa (p12) subunit. Interacts with BIRC6/bruce.

Subcellular Location:
Cytoplasm.

Tissue Specificity:
Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system.

Post-translational modifications:
Cleavage by granzyme B, caspase-6, caspase-8 and caspase-10 generates the two active subunits. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. Active heterodimers between the small subunit of caspase-7 protease and the large subunit of caspase-3 also occur and vice versa.
S-nitrosylated on its catalytic site cysteine in unstimulated human cell lines and denitrosylated upon activation of the Fas apoptotic pathway, associated with an increase in intracellular caspase activity. Fas therefore activates caspase-3 not only by inducing the cleavage of the caspase zymogen to its active subunits, but also by stimulating the denitrosylation of its active site thiol.

Similarity:
Belongs to the peptidase C14A family.

SWISS:
P55213

Gene ID:
836

Database links:

Entrez Gene: 836 Human

Entrez Gene: 12367 Mouse

Entrez Gene: 397244 Pig

Entrez Gene: 100008168 Rabbit

Entrez Gene: 25402 Rat

Omim: 600636 Human

SwissProt: P42574 Human

SwissProt: P70677 Mouse

SwissProt: Q95ND5 Pig

SwissProt: Q8MJC3 Rabbit

SwissProt: P55213 Rat

Unigene: 141125 Human

Unigene: 3885 Mouse

Unigene: 10562 Rat




Picture

Sample:
Lane 1: Spleen (Mouse) Lysate at 40 ug
Lane 2: Lung (Mouse) Lysate at 40 ug
Primary: Anti-Caspase 3 precursor (SL2593R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 32 kD
Observed band size: 32 kD
Sample:
Liver (Mouse) Lysate at 40 ug
Primary: Anti-Caspase 3 precursor  (SL2593R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 28 kD
Observed band size: 28 kD
Sample:
Lane1: Brain (Mouse) Lysate at 30 ug
Lane2: Liver (Mouse) Lysate at 30 ug
Primary: Anti-pro-caspase-3 (SL2593R) at 1:300 dilution;
Secondary: HRP conjugated Goat Anti-Rabbit IgG(SL0295G-HRP) at 1:3000 dilution;
Predicted band size : 28kD
Observed band size : 30kD
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-pro-caspase-3 Polyclonal Antibody, Unconjugated(SL2593R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining
Blank control (blue line): Hela (fixed with 80% methanol (5 min at -20℃) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature).
Primary Antibody (green line): Rabbit Anti-Caspase 3 precursor antibody (SL2593R), Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1μg /test.
Blank control: NIH/3T3.
Primary Antibody (green line): Rabbit Anti-Caspase 3 precursor antibody (SL2593R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF488
Dilution: 1μg /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.
Blank control (blue line): HL60 (fixed with 2% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature).
Primary Antibody (green line): Rabbit Anti-Caspase 3 precursor antibody (SL2593R), Dilution: 1μg /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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