Human/Mouse/Rat p53 (S15) Cell-Based ELISA Kit

p53

-20°C

Adherent Cell Culture

Colorimetric

96-well Microplate

uncoated 96-well Microplate
Wash Buffer A
Wash Buffer B
Fixing Solution
Quenching Buffer
Blocking Buffer
Anti-phospho 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 manufacture'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.

41116158

The antibodies provided in this kit recognizes Human/Rat/Mouse p53 phosphorylated at Serine-15 and total p53 for comparison.

T47D cells were stimulated by 300J/m2 of UV radiation then incubated for 4 hours at 37°C
T47D cells were treated or untreated with 50 J/m2 UV. Cell lysates were analyzed using Western Blot.