Hepcidin-25 peptide (ab31875)

I would like to know in which buffer I should dissolve the Hepcidin-25 peptide (ab31875). Is water ok? Thank you very much.

ANSWER:

Thank you for contacting us.

The peptide should already be in solution. Is that not the case? You should have 100ul. Be sure to centrifuge the tube to pulll down any solution trapped in the threads of the tube.

If you want to further dilute the stock solution, distilled water will be fine.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Do you have information of the reactivity of "Hepcidin-25 peptide (ab31875)" on mouse cells? Or is it possible to obtain a test-sample to test the reactivity?

ANSWER:

Thank you for contacting us.

The product ab31875 is a synthetic 25 amino acid peptide that is not, to our knowledge, active. This peptide has been used to produce the antibody ab75883, which is validated for use on mouse and human samples in WB, IHC-Fr, and ICC/IF. I hope this helps, please let me know if you have any other questions.

We appreciated the advice we have obteined from Abcam, but I communicate that the use, in our experiments, of 12% gels with MES SDS buffer is equivalent to having done running the proteins at in a gel of approximately 18% polyacrylamide, so we are confident that the protein of our interest has not been lost.

Please let me know if you can send the the immunizing peptide for blocking experiments.

ANSWER:

Thank you for your reply.

I am still concerned that the resolving conditions are not suitable for such a small protein. I will send on the peptide as an exception. However, I am not sure how it will be used for blocking if a band of the appropriate size is not detected.

Please confirm the order number and date of shipping so that I can send on the peptide. Looking forward to your reply.

Thanks very much for your detailed response, that’s helpful. Just a couple of further questions/comments: when you state that the peptide sequence corresponds to aas 60-84, are you referring to ab31876 or ab31875 or both?

There is evidence in the literature that pro-hepcidin is found in the blood as well as located intracellularly, and pre-pro-hepcidin can also be secreted from cells.

I do think it might be helpful for customers to know that the ‘hepcidin-25’ antibodies are likely to recognise other forms of hepcidin (since it can be important to distinguish between active hepcidin and inactive precursors), and therefore it would be good to provide this information on the data sheet.

 I hope to place an order for one of the hepcidin peptides and some anti-hepcidin antibodies soon.

Best wishes,

ANSWER:

Thanks you for your email. Dr. Tanya is away this week so I am looking after her inbox.

The peptide sequence for ab31876 corresponds to amino acid XXXX and peptide sequence for ab31875 corresponds to XXXX amino acids.

The antibody cross reactivity always depends on the immunogen sequences, so customers will automatically know from the datasheet e.g. which part of the immunogen the antibody is specific to and in which part of the processed protein, the immunogen lies. This is very obvious information so we do not think we need to publish this information. However we are always happy to answer any inquiry we receive regarding there products.

I hope this will be helpful. Should you any other please does not hesitate to contact us.

Hi, I have a query about this product (hepcidin-25 peptide). I don't understand why this product is termed 'hepcidin-25' when it is actually full length pre-pro-hepcidin rather than the 25 amino acid active form of hepcidin. It's very misleading, can you explain what you mean by 'hepcidin-25' here? I'm also confused about the 'hepcidin-25' antibodies you have - are these specific to the 25 aa form or the 84 aa protein?

Many thanks for your help,

ANSWER:

Thank you for your enquiry and your interest in our products.

I have conducted a quick blast and the peptide sequence picks up both hepcidin and hepcidin 25. The peptide sequence corresponds to aas 60-84 of the full length protein.

The antibody recognizes the processed hepcidin 25 (60-84 aa) which will be found at low concentrations in the blood (presumably) and it will also detect the full length pre-pro protein which would be found in the cell – hence easier to run on a Western blot.

Therefore it would be very difficult to get an antibody specific for the processed protein.

I hope this helps and if I can assist further, please do not hesitate to contact me.