Human/Mouse/Rat Phospho-p38 MAPK (T180/Y182) Cell-Based ELISA

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
H
R
P-
Conjugated Secondary Antibody
T
M
B 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 R
T with shaking.
Add 200 µl of prepared 1
X Quenching Buffer and incubate 20 min at R
T.
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 R
T.
Add 50 µl of prepared 1
X H
R
P-
Anti-
Rabbit or Mouse Ig
G and incubate for 1 h at R
T.
Add 100 µl of T
M
B One-
Step Substrate Reagent to each well.
Incubate 30 min at R
T.
Add 50 µl of Stop Solution to each well.
Read at 450 nm immediately.

Mitogen-activated protein kinase 14 (MAP kinase 14) (MAPK 14) (EC 2.7.11.24) (Cytokine suppressive anti-inflammatory drug-binding protein) (CSAID-binding protein) (CSBP) (MAP kinase MXI2) (MAX-interacting protein 2) (Mitogen-activated protein kinase p38 alpha) (MAP kinase p38 alpha) (Stress-activated protein kinase 2a) (SAPK2a)