Anti-VCP antibody [5] (ab11433)

Dear Mr/Ms,
I am usingAnti-VCP antibody(ab11433)from your company for co-ip purpose. The co-ip kit I am using suggests thatI should use 2ug antibody while there is no concentration in the datasheet. Would you please tell me the concentration or the approximate amount of antibody I should use? Thank you very much!
Best Regards

ANSWER:

Thank you for contacting us.

The Anti-VCP antibody [5] (ab11433) is supplied as unpurified ascites fluid for which we do not quantify the protein concentration. Ascites can range in concentration from 0.5-10 mg/ml IgG but are generally closer to 5 mg/ml. We suggest a standard concentration of 10-50 ug/ml antibody for Immunoprecipitation experiments which would translate to about a dilution of 1:100-1:500.

You may also find it useful to have a look at the Abreviews submitted by other users of this antibody in IP, which you can find at the product’s datasheet under the tab “Abreviews”.

I hope this information is helpful. Please feel free to contact me again with any additional questions you may have.

Dear friends at Abcam,   I am working on a project with a novel VCP/p97 protein interactor and would very much like to test your mouse monoclonal anti-VCP antibody (ab11433) for IP and Immunofluorescence. Currently I work with anti-VCP ab (Novus Biologicals),which is not good for IP, and since it is rabbit polyclonal, I can not combine it with ab against this novel protein we work with (also rabbit poly), for IF staining of endogenous proteins. I would appreciate very much if you could send me a small aliquot of this antibody to test it, and if I find it suitable for my work, I will purchase an aliquot and continue to work with it. Also, we will mention this ab in any publication created with a help of this ab, as well. Thank you in advance and best regards.    

ANSWER:

Thank you for contacting us.

Because we carry over 80,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase. In the case of ab11433 the covered applications include IP and ICC/IF, IHC-P, IHC-Fr, Flow Cyt, WB.

We like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates. To find out more about our Abreview system, please see the following link:

http://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.  

Would you tell me when this antibody used in IP, how to dilute it? Thank you.

ANSWER:

The best dilution factor for the antibody will need to be determined by comparing immunoprecipitated protein with protein left remaining in the "supernatant" after running over a Protein G column. As a starting point, try 5 ul of antibody added to 1 mg of lysate. The concentration of the antibody is not determined since it is an ascites fluid, which can vary from 1 to 10 mg/ml.

I hope this helps. Please let me know if you have any other questions.

I would like to know if these antibodies can detect Cdc48 protein from yeast lysate (S. cerevisiae) ? If YES, which dilution is optimal for WB (1:2000 ? ) Thank you in advance!

ANSWER:

Thank you for your enquiry. All the information we have on species cross reactivity is specified on the datasheet. To our knowledge, cross reactivity with S. cerevisiae has not yet been tested. Should you decide to go ahead and purchase this product, please let us know how you get on and in return we will forward a reward of your choice, typically an Amazon gift voucher.