Caspase 3 ELISA Kit (CASP3)

Full Name: Caspase 3 ELISA Kit (CASP3)
Reactivity: Human
Sample Type: Serum, Plasma, Biological Fluids, Tissue Homogenates
Sensitivity: 0.188 ng/ml

INTRODUCTION

Caspase-3 is an essential caspase protein encoded by the CASP3 gene. The CASP3 gene has wide cross-species comparisons, from mammals to lizards and even teleosts. The gene is located on the long arm of chromosome 7 at position 17.1, it has 9 exons and 8 introns. It encodes a protein which contains 2 zinc fingers, a serine/threonine kinase domain, proline-rich region and cysteine-rich region. The protein is a member of the apoptosis-associated speck-like protein (ASC) family and is an anti-apoptotic factor. CASP3 regulates cell cycle progression, proliferation and apoptosis, via regulation of the Akt signalling pathway. Akt is a kinase enzyme that in humans is encoded by the “AKT1” and “AKT2” genes. It is a serine/threonine protein kinase that was originally found to be associated with cell growth and proliferation.

Caspase-3 are inactive enzymes that need to be processed by specific proteolytic cleavage at specific aspartic residues in order to activate them. Asp-1858, a particular aspartic residue found at the C-terminus, is cleaved by proteolysis to activate them, whereas Asp-1203 is found at the N-terminus. The activated enzyme proceeds with hydrolytic cleavage of Asp-1203 creating a new C/D terminus bond, before eventually joining together to form an active enzyme. CASP3 then proceeds with proteolytic cleavage of various substrates, including poly(ADP ribose) polymerase (PARP). This process is responsible for DNA fragmentation during apoptosis.

INTENDED USE

Human caspase 3 ELISA kit is intended for detecting amount of CASP3 (caspase 3, CPP32, SCA-1, apoptain) present in human serum, plasma, biological fluids and tissue homogenates.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

One 96-Well Plate: Pre-coated with CASP3
Standards: Lyophilized recombinant.
Sample/Standard Dilution Buffer.
Biotinylated-labelled Antibody.
Antibody Dilution Buffer.
HRP-Streptavidin Conjugate (SABC).
SABC Dilution Buffer.
TMB Substrate.
Wash Buffer (25x).
Plate Sealer.
Product Instructions.

TYPICAL RESULTS

The following caspase 3 ELISA kit it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 0.312, 0.625, 1.25, 2.5, 5, 10, 20 ng/ml.
Reactivity: Human
Sensitivity: 0.188 ng/ml
Range: 0.313 – 20 ng/ml
Principle: Sandwich
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for CASP3, no cross reactivity or interference between CASP3 and analogues was detected.
– Recovery: Serum (88 – 104%), EDTA Plasma (85 – 105%), Heparin Plasma (90 – 99%).
– Linearity: Serum (90 – 100%), EDTA Plasma (88 – 96%), Heparin Plasma (80 – 91%).
– Precison Intra-Assay: CV < 8%.
– Precison Inter-Assay: CV < 10%.
– Stability: Less than 10%.

REFERENCES

Caspase 3-mediated stimulation of tumor cell repopulation during cancer radiotherapy. Nat Med. (2011) 17 (7): 860-6. Huang Q., et al.
Hypochlorous acid reversibly inhibits caspase-3: a potential regulator of apoptosis. Free Radic Res. (2020) 54 (1): 43-56. Jeelani R., et al.
Flow cytometric detection of activated caspase 3. Methods Mol Biol. (2008) 414: 47-56. Fox R. and Aubert M.

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