Anti-eIF2B epsilon (phospho S539) antibody (ab4775)

Customer's email is

Thank you very much for your cooperation.

Best

ANSWER:

Thank you for your interest and for passing the customer's details.
DISCOUNT CODE: 42219
Expiration date: [31/05/2012]
I am very pleased to hear you would like to accept our offer and test ab4775 in Mouse. This code will give you 1 free PRIMARY ANTIBODYbefore the expiration date. To redeem this offer, please submit an Abreview for Mouse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.
Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.
Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.
The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Thank you for the reply.

The customer agreed to proceed with the abreview promotionwith ab4775 for WB with mouse samples. Are you interested in this deal in this case? Do you think that there is cross reactivity with mouse?
I highly appreciate your assistance.
Looking forward to your reply.
Thanks again.
Have a nice day,

ANSWER:

Thank you for your response.
I can certainly offer a discount code for testing and this case seems to be stronger since there may be some publication regarding the cross-reactivity with mouse (Western blot application).
Would you be so kind to pass your customer's details so that I can provide the discount as soon as possible?
I look forward to hearing from you soon.

Further to previous correspondence.

thanks a lot for your response and your suggestions. I recommended your suggestion to block with BSA instead of milk powder to reduce background staining but you would expect that the antibody is more specific for Phospho-elF2B epsilon. To answer your questions: the customer used an antibody for elF2B epsilon from Cell Signaling. Cell Signaling offered also a phospho-specific antibody which is now no longer in their sortiment due to a variety of problems. The cells the customer used for WB contain GSK3, also active in non-stimulated cells (see JBC 280, 33006-33014, 2005). elF2B should be phosphorylated under these conditions.

Do you offer a positive control the customer could use for the immunoblotting with this antibody?

Thanks a lot in advance for further support!

Have a nice day and weekend!

Best regards

ANSWER:

Thank you for getting back to me.

Thank you for confirming that both eIF2B and GSK3 are expressed in these cells.

We recommend a positive control of recombinant eIF-2B epsilon treated with GSK 3 beta and lambda phosphatase or CHO-T cells transfected with human insulin receptor (IR) +/- insulin. Unfortunately we do not cell these as lysates or extracts.

I hope this information helps. Please do not hesitate to contact me should you require further assistance.