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Anti-KIF14 antibody (ab3746)

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1 question for ab3746

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Question 1

Wednesday 02-May-2012

Dear Sir/Madam,
Our lab (Adrian Whithouse lab, Leeds University) recent purchased a large batch of antibodies from your company. Purchase Order 4500686042. Two of which are not working correctly.
I have attachted a word document containing Western blots using the two primary antibodies:-
Anti-Periplakin antibody - (ab72422)
Anti-KIF14 antibody - (ab3746)
These western blots have been exposed for 15 minutes. Whereas, the Abcam info sheets state 10 seconds required.
The Periplakin blot shows a band at approximately 55 kDa that appeared after 10 seconds, however the band just below 200 kDa (which I believe to be periplakin) wasn't picked up until 15 minutes exposure.
The KIF14 blots shows a band at just below 200kDa (which I believe to be KIF14), however this wasn't picked up until 15 minutues exposure.
The samples that I used to perform this experiment, are the same samples used to conduct MS/MS-SILAC and the results of this confirmed that there was a drastic increase in these proteins in these samples compared to mock whole cell lysate. Hence I should be seeing a strong band for both KIF14 and Periplakin.
Is there any alternativies or a differnt batch of these antibodies that you can send us?
I look forward to hearing from you.
Kind regards

ANSWER:

 

Thank you for contacting us and for sending an image of your blot.

I am sorry to hear you are experiencing difficulties with two of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I am addingsome questions belowso that we can gather further information regarding the samples tested and the protocol used. Once we receive the answers, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

Samples
Species of the sample :
Type of the tissue/cell lysate :

Transfer
Transfer conditions (time and temperature, wet or semi-dry):
Transfer buffer :
Protein transfer verified?

Blocking
Blocking agent and concentration :

Primary antibody
concentration or dilution :
Diluent buffer :
Incubation time :
Incubation temperature :

I look forward to receiving your reply.

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