Recommended Products
biological source
rabbit
Quality Level
antibody form
purified immunoglobulin
antibody product type
primary antibodies
clone
polyclonal
species reactivity
mouse, human, rat
manufacturer/tradename
Upstate®
technique(s)
immunohistochemistry: suitable
western blot: suitable
isotype
IgG
NCBI accession no.
UniProt accession no.
shipped in
wet ice
target post-translational modification
unmodified
Gene Information
human ... CASP3(836)
General description
Caspase-3 (UniProt: P42574; also known as EC:3.4.22.56, CASP-3, Apopain, Cysteine protease CPP32, CPP-32, Protein Yama, SREBP cleavage activity 1, SCA-1) is encoded by the CASP3 (also known as CPP32) gene (Gene ID: 836) in human. Cysteine-aspartic proteases or Caspases play essential roles in apoptosis, necrosis, and inflammation. Historically, caspases were numbered in the order in which they were identified. Caspase-3 is a heterotetrameric enzyme that consists of two anti-parallel arranged heterodimers, each one formed by a 17 kDa (p17) and a 12 kDa (p12) subunit. Caspase-3 is initially produced with a propeptide sequence (aa 1-9), the removal of which yields the 268 aa. caspase-3 proenzyme. Upon activation, the proenzyme is proteolytically cleaved first between Asp175-Ser176 to generate a p20 (aa 10-175) fragment and the p12 (aa 176-277) subunit. Further cleavage of the p20 fragment between Asp28-Ser29 produces the p17 (aa 29-175) subunit. The p17 and p12 subunits dimerize and forms the active caspase-3 enzyme. Caspase-3 has a strict requirement for an Asp residue at positions P1 and P4. It has a preferred cleavage sequence of Asp-Xaa-Xaa-Asp-|- with a hydrophobic amino-acid residue at P2 and a hydrophilic amino-acid residue at P3, although Val or Ala are also accepted at this position. Caspase-3 is 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 Asp216-|-Gly217 bond. Caspase-3 mediates the proteolytic activation of caspases-6 and -7, while caspase-3 itself is processed and activated by caspase-8, -9, or -10.
Specificity
Recognizes full-length Caspase 3 (Yama/Apopain) and proteolytic fragments.
Immunogen
Human full-length Caspase 3 fusion protein containing a histidine-6 tag
Application
This Anti-Caspase 3 Antibody is validated for use in Immunihistochmistry and Western Blotting for the detection of Caspase 3.
Western Blotting Analysis: 1μg/mL of this antibody detects Caspase-3 in A431 cell lysate.
Immunohistochemistry (Paraffin) Analysis: A 1:250 dilution of this antibody detected Caspase-3 in Human tonsil tissue sections.
Immunohistochemistry (Paraffin) Analysis: A 1:250 dilution of this antibody detected Caspase-3 in Human tonsil tissue sections.
Quality
routinely evaluated by immunoblot on RIPA lysates from non-stimulated human A431 cells, mouse 3T3/A31 or rat PC12 cells
Target description
32 kDa
Linkage
Replaces: 04-1090; 04-439
Physical form
Format: Purified
Protein A purified IgG in of 0.1M Tris-glycine, pH 7.4, 0.15M NaCl,and 0.05% sodium azide.
Storage and Stability
Stable for 2 years at 2-8°C from date of shipment. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
Analysis Note
Control
Positive Antigen Control: Catalog #12-301, non-stimulated A431 cell lysate. Add 2.5µL of 2-mercaptoethanol/100µL of lysate and boil for 5 minutes to reduce the preparation. Load 20µg of reduced lysate per lane for mingels.
Positive Antigen Control: Catalog #12-301, non-stimulated A431 cell lysate. Add 2.5µL of 2-mercaptoethanol/100µL of lysate and boil for 5 minutes to reduce the preparation. Load 20µg of reduced lysate per lane for mingels.
Legal Information
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
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Certificates of Analysis (COA)
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M Leist et al.
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