Custom Human/Mouse Cleaved-Caspase-3 (D175) Cell-based ELISA Kit

Blue ice

RayBio® CASP-3 (D175) Cell-based ELISA Kit. This ELISA is for measuring cleaved CASP-3 (Asp-175) in human adherent cell lines.

Uncoated 96-well Strip Microplate
Wash Buffers
Fixing Solution
Quneching Buffer
Blocking Buffer
Anti-cleaved Antibody
Anti-pan antibody
HRP-Conjugated Secondary Antibody
TMB One-Step Substrate
Stop Solution

Seed 10, 000-30, 000 cells into each well and incubate overnight.
Apply various treatment, inhibitors or activators according to manufacturer's instructions.
Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking.
Add 200 µl of prepared 1
X Quenching Buffer and incubate 20 min at RT.
Add 200 µl of Blocking Solution and incubate for 1 h at 37°C.
Add 50 µl of 1
X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.
Add 50 µl of prepared 1
X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.
Add 100 µl of TMB One-Step Substrate Reagent to each well.
Incubate 30 min at RT.
Add 50 µl of Stop Solution to each well.
Read at 450 nm immediately.

Caspase-3 (CASP-3) (EC 3.4.22.56) (Apopain) (Cysteine protease CPP32) (CPP-32) (Protein Yama) (SREBP cleavage activity 1) (SCA-1) [Cleaved into: Caspase-3 subunit p17 Caspase-3 subunit p12]