Human Phospho-TrkB (Y816) and Total TrkB ELISA Kit

Assay Kits|Phosphorylation Assays|ELISA Kits|Phosphorylation ELISA|PTM ELISA

-20°C

Cell Lysates
Tissue Lysates

Colorimetric

96-well Microplate

Pre-Coated 96-well Strip Microplate
Wash Buffer
Anti-Phospho Antibody
HRP-Conjugated Secondary Antibody
Assay Diluent
TMB One-Step Substrate
Stop Solution
Lysis Buffer
Positive Control Sample

Prepare all reagents and samples as instructed in the manual.
Add 100 µl of sample or positive control to each well.
Incubate 2.5 h at RT or O/N at 4°C.
Add 100 µl of prepared primary antibody to each well.
Incubate 1 h at RT.
Add 100 µl of prepared 1
X HRP-Streptavidin to each well.
Incubate 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

This ELISA kit recognizes Human TrkB phosphorylated at Tyrosine-816 and Total TrkB.

RayBio® Phospho-TrkB (Tyr816) and Total TrkB ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in Human cell lysates. By determining phosphorylated TrkB protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of Human phospho-TrkB and total TrkB. An anti-pan TrkB antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and TrkB present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho-TrkB (Tyr816) antibody is added to detect phosphorylated TrkB. In the remaining wells, biotinylated anti-pan-TrkB antibody is used to detect pan TrkB. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or HRP-Streptavidin is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of TrkB (Tyr816) or pan TrkB bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

U-251 cells were treated with Pervanadate. Cell lysates were analyzed using this phosphoELISA and Western Blot.