The epidermal growth factor receptor (EGFR; ErbB-1; HER1) is a receptor tyrosine kinase critical for regulating cell proliferation, differentiation, survival, migration, and organ development. Dysregulation of EGFR signaling-through mutations, overexpression, or loss-of-function, is implicated in a diverse spectrum of human diseases.
Tribioscience’s Fast EGFR Kinase Activity Assay is designed to quantitatively detect EGFR Kinase Activity levels in different tissues including skin, muscle, neural, serum, and other biological samples. In this assay EGFR phosphorates tyrosine-based substrate in the presence of co-substrate ATP. The phosphotyrosine concentration can be detected with anti-phorsphotyrosine based sandwich ELISA technology. The assay combines multiple reactions into a one-step. The measurement can be finished in 1.5 hours. The detection range is from 78 to 5000 pg/mL. The levels of sample EGFR Kinase activity are parallel to the standard curves obtained using the kit standards linearly. These results indicate that this kit can be used to determine relative mass values for natural EGFR Kinase Activity.
MAIN FEATURES
Simple: Multiple reactions into one-step.
Accurate: Directly measure the phorsphoration of tyrosine
Fast: Just 1.5 hours.
KIT CONTENT AND STORAGE CONDITIONS
PART
PART#
DESCRIPTION
STORAGE OF OPENED/ RECONSTITUTED
Substrate coated plate
TBS2048A
96 well polystyrene microplate (12 strips of 8 wells) coated with a specific substrate for EGFR kinase.
Return unused wells to the foil pouch. Reseal along entire edge of the zip-seal. May be stored for up to 1 month at 2-8 °C.
Co-substrate
TBS2048B
50 µL of ATP (50x)
Aliquot and store at -20 ℃ for up to 1 month in a manual defrost freezer. Avoid repeated freeze-thaw cycles.
EGFR Standard
TBS2048C
20 µL of EGFR kinase (0.125 ng/µL).
Activate Buffer
TBS2048D
6 mL of a buffer with activators.
Detection A
TBS2048E
12 µL of anti-mouse phosphotyrosine antibody (200x).
Detection B
TBS2048F
6 µL of anti-mouse IgG-HRP.
May be stored for up to 6 months at 2-8 °C.
Assay Diluent
TBS2048G
15 mL of a buffered protein base with preservatives.
Wash Buffer
TBS2048H
12 mL of concentrated solution (10x).
TMB Substrate
TBS2048I
11 mL of ultra-sensitive TMB substrate.
Stop Solution
TBS2048J
6 mL of 2 N sulfuric acid.
Store the unopened kit at -20 ℃. Do not use past kit expiration date.
The kit contains sufficient materials to run an ELISA on one 96 well plate.
