Application
Western blot
Sample
Human Cell lysate - whole cell (Prostate, Kidney)
Gel Running Conditions
Reduced Non-Denaturing (Native) (10)
Loading amount
40 µg
Specification
Prostate, Kidney
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Incubation time
15 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 0.1% TBS-T
Dilution
1/1000
Secondary antibody
Dilution
1/2000
Name
Non-Abcam antibody was used: Anti-Rabbit Secondary
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Negative control
N/A
Exposure
10 minute(s) and 0 second(s)
Bands
Specific: none kDa Non-specific: 30, 45, 50, 55, 60, kDa
Positive control
I used total protein lysates of HEK293T cells, as suggested by Abcam.
Additional data
Additional Notes
This antibody performed very poorly for me. While the background was relatively clean, there was just too many non-specific bands around the predicted area of interest for me to determine which was the real band. This was in contrast to the pictures that are provided on Abcam's website, which showed one clear band at the correct size. I loaded both 20ug and 40ug of both my test sample and the positive control. While there does seem to be bands present in the predicted 45kDa region, there are too many other bands there, making it impossible to confidently determine the correct one. Unfortunately, there was no other lot in stock at the time of this review, so I don't know if it's just a bad lot. Abcam customer service, however, was extremely helpful and refunded me. Thanks!
Abcam response
We are very sorry that this antibody did not work well in this case. We have followed up with the customer in accordance to our AbPromise (www.abcam.com/abpromise).
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
投稿 Nov 14 2013