Anti-ADAM15 antibody (ab84834)

Dear Sir or Madam,

I would like to enquire about a possible cross-reaction of one the abcam antibodies we use in our lab. I use the anti-ADAM15 antibody (ab84834) to detect ADAM15 by western blotting after overexpression in HEK293 cells. In my experiments I wasn't also seeing signals for ADAM15 but also two very distinct bands in HEK293 cells which were overexpressing ADAM12. These bands run at approx. 100 and 75 kDa which would fit to the predicted size of the pro- and active form of ADAM12. With this result the question rises if there's there any known cross-reactivity for
this antibody?

Attached to this e-mail you'll find a picture of the western blot

Thank you for your effords.

Best regards

ANSWER:

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for WB and human samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I would appreciate if you are also able to provide any image which would help us to assess the results.

Adam 15 has 11 isoforms according to the Uniprot protein database site:
http://www.uniprot.org/uniprot/Q13444

I have blasted the sequence of the largest isoform, and this does not indicate anysignificantalignment with the Adam 12 protein, so there should not be crossreactivity with Adam 12.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.




Order Details
Antibody code:

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number:



General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, wrong band size, more bands, no band etc.)


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


Amount of protein loaded


Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method (ECL, ECLPlus etc.)


Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the Western?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?


Additional Notes:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.