Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T cells)
Gel Running Conditions
Reduced Denaturing (4-12% gel)
Loading amount
10 µg
Specification
HEK293T cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C
Dilution
1/1000
Secondary antibody
Dilution
1/10000
Name
Non-Abcam antibody was used: anti-mouse IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL Prime
Negative control
Mock transfected cells
Exposure
3 second(s)
Bands
Specific: 23 kDa Non-specific: 23 kDa
Positive control
Cells transfected with plasmid encoding Nef and its mutants.
Additional data
Additional Notes
This antibody fails to detect Nef expressed in transfected cells. It cross-reacts with an unknown cellular protein of the same molecular weight (mock transfected cells as negative control) as well as the myoglobin red band of the molecular weight marker (SeeBlue Plus 2, Thermo Fisher).
Abcam response
Our Scientific Support team has resolved the issue in accordance with our Abpromise.
A free of charge replacement with the alternative ab42358 was sent and gives positive results.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Dr. Jason Lee
Verified customer
投稿 Aug 15 2016