For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome
If you continue without changing your cookie settings, we'll assume you’re happy with this.
I had a customer contact me regarding some problems with AB7815 in WB and IHC (no signal). I realize that this antibody is not longer validated for paraffin sections, but the customer also tried it in WB without success. This item was ordered on PO xxxx. Can you please take a look at the customer's protocols attached?
Asked on Jan 31 2012
Thank you and your customer for taking the time to complete our questionnaire and contact us. I am sorry to hear your customer has had difficulty obtaining satisfactory results from this antibody. I can confirm that ab7815 is covered by our Abpromise guarantee when used in WB.
The details your customer has kindly provided will enable us to investigate this case for your customer and this is also helpful in our records for monitoring of quality.
Reviewing this case, I would like to offer some suggestions to help optimise the results from ab7815:
1.) I suggest use a positive control: Fresh skin biopsy (small fresh skin biopsy incubated for 4 hours with TNF (1 ng/ml), IL-1 (100 u/ml) or LPS (1 ìg/ml) in tissue culture medium at 37oC).
2.) I recommend to use reducing and denaturing conditions: add SDS to the lysis puffer and add beta mercaptho-ethanol to the loading buffer.
3.) Blocking in BSA can significantly increase the signal of some antibodies. Therefore I suggest to use 5% BSA as blocking agent.
4.) Since there is no background I suggest to not use any blocking agent in the antibody solution and to dilute the antibody less. An other customer had good results with a dilution of 1:500.
We are happy to offer this technical support. In the event that a product is not functioning in the applications 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 hope this information is helpful, thank you and your customer for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.
Answered on Jan 31 2012