Application
Western blot
Sample
Human Cell lysate - whole cell (A549, HEK293)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
50000 cells
Specification
A549, HEK293
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% BSA in TBS
Dilution
1/500
Secondary antibody
Dilution
1/2000
Name
Non-Abcam antibody was used: Goat anti-rabbit HRP IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: 108 kDa Non-specific: 50-60, ~47 (HEK293) kDa
Positive control
A549, HEK293 cell lysates.
Additional data
Additional Notes
Also tested with 5% milk as a blocking agent, but only non-specific bands at 50-60kDa were detected.
Abcam response
Thank you for sharing this data with us. ab64820 has previously been tested in western blot on HEK 293 cell lysates and showed a band 108kDa (please see datasheet). However, this abreview shows high background at ~50kDa, under these conditions. Abcam would welcome feedback from other customers using this product.
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.
Abcam user community
Verified customer
提交于 Mar 28 2014