Human Phospho-TAK1 (Ser412) and Total TAK1 Cell-Based ELISA Kit

Blue ice

Adherent Cell Culture

Colorimetric

96-well Microplate

Uncoated 96-well Strip Microplate
Wash Buffers
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 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.

Mitogen-activated protein kinase kinase kinase 7 (EC 2.7.11.25) (Transforming growth factor-beta-activated kinase 1) (TGF-beta-activated kinase 1)

This ELISA kit recognizes Human TAK1 phosphorylated at site Serine-412 as well as total TAK1.

HeLa cells were treated with Calyculin A and IL-1 beta