Anti-EGF antibody [EGF-10] (ab10409)

We see no signal or non-specific epidermal staining on sections of paraffin-embedded human skin.

ANSWER:

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number ***** for ab9695.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

What are the protocol conditions for IHC-P?

ANSWER:

Thank you for contacting Abcam regarding ab10409.

I have confirmed the following protocol was used in IHC-P:

Heat Induced Epitope Retrieval (HIER) is performed by heating the slides immersed in retrieval solution: 10 mM sodium citrate buffer, pH 6.0, with 1 mM EDTA, at 125 °C for 4 minutes in a pressure boiler. After boiling is completed, slides remain in the pressure boiler and are allowed to cool down to 90 °C. The total processing time is ~45 minutes.

Staining was performed using a Lab Vision Autostainer 480.  All incubations were performed at RT and all reagents are applied at a volume of 300ul.

1.  Rinse in wash buffer - PBS containing 0.2% Tween 20 2.  Incubate with Ultra V Block 5 minutes 3.  Rinse 2x in wash buffer 4.  Incubate with primary antibody diluted 1:500 for 30 minutes 5.  Rinse 3X in wash buffer 6.  Incubate with HRP labeled polymer for 30 minutes 7.  Rinse 2X in wash buffer 8.  Develop using DAB 9.  Rinse 2X in distilled water 10.  Counterstain

I hope this information is helpful.  Please do not hesitate to contact me if you have any additional questions.

I would like to have a positive control for EGF ab10409 for western blots, do you have any suggestions?

ANSWER:

Thank you for your enquiry. We tested ab10409 with a recombinant EGF sample, which is available from Sigma I believe, catalogue number E9644.

Please do not hesitate to contact me again if you require further information,

I am currently working on neural stem cells. I wish to assess the EGF antibody [EGF-10] (ab10409) for immunoprecicpitation of rat EGF.I would appreciate to receive a free trial size of your antibody. Yours sincerely

ANSWER:

Thanks for your enquiry regarding ab10409. Unfortunately we do not currently offer free trial size batches of antibodies to test.

I should take this opportunity to mention that this antibody may not recognise rat EGF since the antibody is known not to recognise mouse EGF. Good luck with your research!

We have made more trials with the new vial of EGF-10 antibody that you sent us but it just does not work. We tried human brain on Western Blot until 40 ug, with an standard protocol. We have not tried EGF positive control as the total human protein is quite expensive and we don't have it, but it is reported that human brain expresses EGF. This trials have taken us quite a long time, and we do not want to continue trying this antibody. We would rather sent it back to you. Please, inform me about our posibilities in this situation.

Sincerely,

ANSWER:

Thank you for your note. We are very sorry to hear that you are still struggling with this antibody and even the replacement vial does not perform as it should do.

We can offer you either a credit note or a full refund; please do let us know how to proceed. We would be grateful if you could confirm your Purchase Order Number or Abcam Order Number.

We apologize for any inconvenience caused and we look forward to hearing from you soon.