Anti-EGF antibody (ab9695)

Thank you very much for your inquiry.
The immunogen for ab9695 was the full length recombinant hEGF (human Epidermal Growth Factor) and it therefore impossible to determine an epitope.
It is possible to determine experimentally if the antibody blocks the binding site of EGFR, but this has not been done and also would have to be repeated with every new immunization round (for every batch of antibody).
To our knowledge, ab9695 has not been tested in immunoprecipitation and therefore is also not covered by our guarantee for this application. However by participating in our AbTrial program you can now use our products in an untested application or species without financial risk.
Simply follow these easy steps below to apply for our AbTrial Program:
1. Reply to this email, letting us know you are interested in testing this product.
2. Our scientists will email you an inactive personal discount code for the value of the product.
3. Purchase and test the product at the regular price.
4. Submit your results, including your discount code in the additional notes section of your Abreview.
5. Once the Abreview is submitted, the discount code will become active.
6. Apply your discount code on your next order to receive that value off.
Please let me know if you have any questions about this offer and I would be happy to help you further.
The Terms and Conditions of this offer can be found at: www.abcam.com/abtrial.

I am sorry for the delay in getting back to you in regards to the antigen retrieval protocol used with the anti-EGF antibody (ab9695).
Please find the protocol attached. Due to the type of tissue used in this testing of ab9695 (human skin tissue), the HEIR would not have been suitable as it can causes the skin to lift off of the slide. This does not mean that it will not be suitable with other tissue samples.
I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

Thank you for contacting us yesterday and your interest in our products.
As discussed over the phone I have been looking into what may serve as suitable positive controls with the anti-EGF antibody (ab9695). As discussed over the phone, we have used the antibody to stain normal human skin as well as murine forepaw vessel. The protein seems to also be expressed in kidney, salivary gland, cerebrum and prostate, so if you have access to any of these tissues they could serve as a control.
If I can be of any further assistance, please do not hesitate to let me know.
Until then, I wish you all the best with your research.

Thank you for your answer.

We guarantee the ab9695 for WB under either reducing or non-reducing conditions. The free of charge replacement will also be covered by our Abcam guarantee so if you would experience problems we would be pleased to help in this case as well.

Please let me know about your decision in this case. i look forward hearing from you if you decide to receive the ab9695 as a replacement.

Thank you for contacting us.
I am sorry to hear that the antibodies are not working as expected.
As we discussed over the phone, if you please could try the no primary control, this would help us a lot in determining whether the problem comes from the secondary antibody alone, or if there are issues with the primary antibody and isotype control as well.
Please let me know your results.
I look forward to hear back from you and assist you further in resolving his problem.

Thakn you for your enquiry. I can confirm the following procedure was used to laboratory test this antibody on skin samples to provide the image on the datasheet. Please note this should be a guideline only and may require further optimization in your laboratory: Antibody concentration: 1.0 μg/ml Tissue: Normal Human Skin (formalin fixed, paraffin embedded) Protocol: Rabbit HRP Polymer Kit (AEC chromogen) Antigen Retrieval: Proteinase K Incubation Times: 10 min Proteinase K 20 min 1X PBS/ 0.2% Trinton X 100 30 min Fc Receptor Block 10 min Sniper Protein Block Overnight Primary Antibody @ 4°C 10 min Peroxidase Block 20 min Rabbit Probe 20 min Rabbit Polymer HRP 5-10 min Romulin AEC chromogen Results: This antibody stained normal skin with 4+ apocrine gland epithelial and secretory cells. Everything else is negative. Ideal staining is at 1.0 μg/ml, yet this antibody concentration range was tested from 1.0 μg/ml- 0.25 μg/ml. I can confirm that any publications that we are currently aware of will be listed on the datasheet. I'm sorry there are none to provide on this occasion for ab9695 EGF antibody. In many cases, regrettably we do not have access to articles mentioning our products, or subscriptions to the journals that publish them. I hope this will be helpful. Should you have any further questions, please do not hesitate to contact us.

Thank you for your enquiry. I have been in touch with the source of this antibody and I can tell you that ab9695 has been tested and shown to neutralise the specific activity of the antigen (EGF). The neutralisation is measured by ND50. ND50 is the amount of antibody to neutralize 50% of the biological activity of the specific concentration of antigen. Unfortunately this antibody is not supplied conjugated to flurophores. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.