Western blot abreview for Anti-FCGRT/FCRN antibody

Good
Abreviews
Application
Western blot
Sample
Mouse Cell lysate - whole cell (MDCK cell lysate transfected with human, mouse and)
Gel Running Conditions
Reduced Denaturing (12% Tris-HCl)
Loading amount
10 µg
Specification
MDCK cell lysate transfected with human, mouse and
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Other product details

Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C
Dilution
1/200

Secondary antibody

Dilution
1/20000
Name

Detection

Detection method
ECL
Negative control
Untransfected MDCK cell lysate was used as negative control. No band was observed.
Exposure
3 minute(s) and 0 second(s)
Positive control
The positive control MDCK cell lysate transfected with human FcRn-YFP expression construct demonstrates a specific band at ~ 70kDa in agreement with the expected molecular weight of ~65kDa. Also the human FcRn protein levels increase when co-transfected with either human, mouse or rabbit beta-2 microglobulin in agreement with previous reports on the necessity of beta-2 microglobulin in the stabilization and proper function of FcRn.

Additional data

Additional Notes
This antibody works well for detection of human FcRn in transfected cells lysates. However, this antibody DOES NOT cross react with mouse FcRn. Beta-actin was used as a loading control. PVDF membrane, cell lysis buffer was PBS, 0.5% CHAPS, + complete protease inhibitor tablet.
Abcam response
We would like to thank this user for their valuable feedback. The homology with mouse is only 65% and we therefore would not expect this antibody to cross-react with mouse.

Jonathan

Verified customer

Submitted Aug 13 2013

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