Western blot abreview for Anti-DDDDK tag antibody - (Equivalent to FLAG antibodies from Sigma)

Poor
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing (4-20)
Loading amount
30 µg
Treatment
transfected with plasmid expressing FLAG
Specification
HEK293
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Other product details

Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C
Dilution
1/1000

Secondary antibody

Dilution
1/5000
Name

Detection

Detection method
ECL prime
Negative control
cell transfected with plasmid without FLAG tagged protein
Exposure
1 second(s)
Bands
Specific: none kDa Non-specific: 70, 35 kDa
Positive control
cell transfected with plasmid expressing FLAG tagged fusion protein

Additional data

Additional Notes
I did western blot on the same lysate with Sigma FLAG antibody (mouse) and worked beautifully. Of course for that I used an anti-mouse secondary antibody. But the bottomline is, this FLAG antibody from abcam clearly does not work.
Abcam response
The reviewer has used this antibody in a tested species and application. The poor performance in this case may be due to lot-to-lot variation. We have offered a refund or replacement as per our Abpromise guarantee.

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Submitted Sep 08 2018

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