Anti-E2F2抗体[2302C4a] (ab70731)

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.

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 ab70731.

The image indicates that there is not signal when using this antibody. The background bands might vanish as soon as the antibody finds a specific protein to bind to.

1.) The trizol protein extraction protocol is using a very high comncentration of urea (10M).

However, urea and thiourea can hydrolyze to cyanate and thiocyanate, respectively, which can modify amino groups on proteins, (e.g. carbamylation of proteins by isocyanate).

I am aware that your cutomer might only have small samples and is using the Trizol for this reason to get protein and RNA/DNA at the same time.

Therefore I suggest to use a positive control with a standard RIPA buffer to optimised the protocol for this antibody first.

2.) When testing our antibodies, our lab uses 5% BSA as a blocking reagent, so I recommend switching to this instead of milk. Some antibodies give stronger, more specific signals on blots blocked with BSA instead of milk, so doing this may improve the results you are seeing, and reduce the non-specific bands. An example of the above is the western blot obtained with the Abcam GAPDH antibody ab9385 : https://www.abcam.com/ab9385 .

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.

Thank you for bringing this to our attention. Can you please describe your samples and protocol? In particular, what is the species and tissue source of the sample, and how was the lysate prepared? What type of membrane are you transferring to from the gel, and how is the membrane blocked before incubation with the antibody? Have you stained blots of your samples successfully with any other antibodies, using these same reagents? I look forward to your reply.