Western blot abreview for Anti-Insulin Receptor (phospho Y972) antibody

Poor
Abreviews
Application
Western blot
Sample
Chinese hamster Cell lysate - whole cell (CHO Expressing Human Insulin Receptor Þ)
Gel Running Conditions
Reduced Denaturing (Biorad TGX Stain-Free, 10%)
Loading amount
30 µg
Treatment
10 nM Insulin for 10min
Specification
CHO Expressing Human Insulin Receptor Þ
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Other product details

Incubation time
9 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 3% BSA in TBST
Dilution
1/5000

Secondary antibody

Dilution
1/10000
Name
Non-Abcam antibody was used: Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L)
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase

Detection

Detection method
ECL+
Negative control
CHO-hIR fasting in F12 for 2h and another 10min.
Exposure
29 second(s)
Bands
Specific: 95 kDa
Positive control
CHO-hIR fasting in F12 for 2h and stimulated with 10 nM Insulin (Sigma) for 10min.

Additional data

Additional Notes
I've repeated for many times, including changing blocking buffer to 5% BSA. Anyway, it's extremely confused that the pY1162/1163 is different from pY972, as the phosphorylation of tryosin residue in site 972 always keeps up with phosphorylation of these three tyrosine residues in pY1158/1162/1163.
Abcam response
Our Scientific Support team is currently investigating this negative Abreview, and we will resolve the issue in accordance with our Abpromise Guarantee.
Username

Dr. yang lou

Verified customer

提交于 Jul 17 2017

注册