产品资料

RayBiotech/Human/Mouse/Rat Phospho-EGFR (Y992) Cell-Based ELISA/1 Plate Kit/CBEL-EGFR992-1

Antigen Information

Accession Number:

P00533

Gene ID:

1956

Gene Symbols:

EGFR
ERBB
ERBB1
HER1

Protein Name & Synonyms:

Epidermal growth factor receptor (EC 2.7.10.1) (Proto-oncogene c-ErbB-1) (Receptor tyrosine-protein kinase erbB-1)

Assay Format

Pathway:

HER/ErbB Signaling

Specificity:

The antibodies provided in this kit recognizes human, mouse and rat EGFR phosphorylated at site Tyrosine-992 and total EGFR for comparison.

Number of Targets Detected:

Species Detected:

Human

Compatible Sample Types:

Adherent Cell Culture

Design Principle:

Cell-based

Method of Detection:

Colorimetric

Quantitative/Semi-Quantitative:

Semi-Quantitative

Solid Support:

96-well Microplate

Product Specifications

Size:

1, 2, or 5 x 96-Well Microplate Kit

Product Features

Site and signal pathway-specific
In vitro detection of adherent cell culture
No sample lysis is needed
Compatible with a standard ELISA plate reader
Faster results than with ELISA
Adaptable for high-throughput screening and drug discovery

Application Notes

Kit Components

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

Other Materials Required

Distilled or deionized water
100 ml and 1 liter graduated cylinders
Tubes to prepare sample dilutions
Protease and Phosphatase inhibitors
Precision pipettes to deliver 2 µl to 1 ml volumes
Adjustable 1-25 ml pipettes for reagent preparation
Benchtop rocker or shaker
Microplate reader capable of measuring absorbance at 450 nm

Protocol Outline

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 1X 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 1X 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 1X 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.

Storage/Stability

The entire kit may be stored at -20^°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.

No posts found

Write A Review

Manual

SDS