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Active Caspase Activated DNase (CAD)

DFF-B; DFFB; CPAN; DFF40; DFF2; DNA Fragmentation Factor 40kDa Beta Polypeptide; Caspase-activated deoxyribonuclease; Caspase-activated nuclease

ACTIVITY TEST

Caspase Activated DNase (CAD), corresponding to product code RPB058Hu01, is a key nuclease in eukaryotic apoptosis. Normally, it exists in an inactive state in the cytoplasm, bound to its inhibitor. Upon apoptotic signal activation, effector caspases (e.g., Caspase-3) cleave the inhibitor, releasing active CAD. The activated enzyme translocates to the nucleus, where it cleaves chromosomal DNA into nucleosome-sized fragments, a hallmark of apoptotic cell death. Its strict regulation prevents unintended DNA damage, ensuring genomic stability in healthy cells. CAD interacts with DFFa (product code L426Hu) as its specific inhibitor; DFFa binds CAD to inhibit its nuclease activity in non-apoptotic cells.Thus a functional ELISA assay was conducted to detect the interaction of recombinant human CAD and recombinant human DFFa. Briefly, CAD was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to DFFa-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-CAD pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630nm immediately. The binding activity of recombinant human CAD and recombinant human DFFa was shown in Figure 1, the EC50 for this effect is 0.01067µg/mL.

USAGE

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Catalog No. Related products for research use of Homo sapiens (Human) Organism species Applications (RESEARCH USE ONLY!)
RPB058Hu01 Recombinant Caspase Activated DNase (CAD) Positive Control; Immunogen; SDS-PAGE; WB.
APB058Hu01 Active Caspase Activated DNase (CAD) Cell culture; Activity Assays.
PAB058Hu01 Polyclonal Antibody to Caspase Activated DNase (CAD) WB; IHC; ICC; IP.
MAB058Hu22 Monoclonal Antibody to Caspase Activated DNase (CAD) WB; IHC; ICC; IP.
SEB058Hu ELISA Kit for Caspase Activated DNase (CAD) Enzyme-linked immunosorbent assay for Antigen Detection.
CMB058Hu Multiplex Assay Kit for Caspase Activated DNase (CAD) ,etc. by CBA (Cytometric Bead Array) FLIA Kit for Antigen Detection.
LMB058Hu Multiplex Assay Kit for Caspase Activated DNase (CAD) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.