Western blot abreview for Anti-Eph receptor B4 antibody

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Abreviews
Application
Western blot
Loading amount
50000 cells
Gel Running Conditions
Reduced Denaturing (10% gel)
Sample
Human Cell lysate - whole cell (A549, HEK293)
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

Name
Non-Abcam antibody was used: Goat anti-rabbit HRP IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/2000

Detection

Detection method
ECL
Bands
Specific: 108 kDa Non-specific: 50-60, ~47 (HEK293) kDa
Exposure
5 minute(s) and 0 second(s)
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.

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Verified customer

Submitted Mar 28 2014

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