CaspSCREEN Caspase Screening Kit

Antigen: Caspase (CASP)

Synonyme: DCP-1, DCP1, l(2)01862, l(2)02132, DmelCG5370, CG5370

Applikation: Enzymaktivitätsmessung (EAA)

Produktnummer: ABIN411789

Produktbeschreibung

Weitere Bezeichnung: Caspase

Beschreibung: Activation of members of caspase-family proteases plays a key role in apoptosis. The CaspSCREEN TM Flow Cytometric Apoptosis Detection Kit provides a convenient means for detecting activation of caspases by flow cytometry in living cells. The assay is based on the cleavage of (aspartyl) 2 -Rhodamine 110 (D 2 R), a reported substrate for members of caspase family proteases. The caspase substrate D 2 R is non-fluorescent, however, upon cleavage of the substrate by cellular caspases, the released rhodamine 110 gives rise to fluorescence that can be measured at excitation of 488 nm and emission of 530 nm. As the D 2 R is more cell-permeable than other fluorometric caspase substrates, activation of caspases can easily be measured in intact cells by flow cytometry.

Anwendungen

Protokoll: 1. Induce apoptosis in cells by desired method. Concurrently incubate a control culture without induction. 2. Count cells and pellet 1 x 10 5 cells. 3. Resuspend cells in 0.3 ml of D 2 R Incubation Buffer. 3. Add 3 μ l of the 1 M DTT (10 mM final concentration). 4. Add 1 μ l of the D 2 R Reagent. 5. Incubate at 37 o C for 10-20 min in the dark. 6. Analyzing cells by flow cytometry using FL-1 channel (Ex/Em = 488/530 nm).

Lagerung: Store kit at –20 o C (Store the Incubation Buffer at 4 o C after opening). • All reagents are stable for 1 year under proper storage conditions. V. References: 1. Hug, H., et al. (1999) Biochemistry 38:13906-13911. 2. Los, M., et al. (1999) Methods in Mo. Biol., Vol. 12, Ch. 14. Human Press Inc., Totowa, NJ.

Beschränkungen: Nur für Forschungszwecke einsetzbar