Search and Browse
Download
Enter Data
|
Assay Method Information |
|
| | In vitro OSR1 Phosphorylation Assay (OSR1) |
| Description: | 0.2 nM GST-hWNK1(1-491) (Carna Biosciences, cat# 05-179); 3.5 ug/ml GST-OSR1; 20 mM HEPES; 0.01% Tween; 5 mM MnCl2; 1 mM TCEP; 2% DMSO; 0.5 uM ATP, pH 7.3 (total volume 40 ul) were incubated for 15 min at room temperature. Samples were 2168then mixed with Laemmli sample buffer and resolved on a 10% Tris-glycine gel, and transferred to nitrocellulose. Membranes were blocked in 5% non-fat milk dissolved in Tris-buffered saline with tween (TBST) for 30 min, washed 3x for 10 min in TBST and incubated with phospho-OSR1 antibody (1:1,000; see Reagents section) at 4 °C overnight. Blots were again washed with TBST and incubated with HRP-conjugated anti-rabbit IgG (1:2,500; catalog # 7074, Cell Signaling) at room temperature for 30 min. Blots were again washed and developed with SuperSignal West Femto Maximum Sensitivity Chemiluminescent Substrate for HRP (Thermo Scientific), and quantified with a Fujifilm LAS-3000 Luminescent Image Analyzer. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |
|
Home |
| |
Search |
| |
Deposit |
| |
SiteMap |
| |
About us |
| |
Email us |
| |
Info |
|
©2000 BindingDB. All rights reserved. |
|||||||||||||