Anti-CCR5 antibody [E164] (ab32048)

Dear Sir/ Madam, Sorry to disturb. Our customer raises an enquiry about ab32048: Anti-CCR5 antibody [E164]. She has purchased in December last year for original testing in WB. Recently, she decides to test it in IHC with human sample unfortunately, no Abpromise is included in IHC. Therefore, she sincerely would like to apply the discount code to test. Would you please help create discount code for her?

ANSWER:

Thank you for contacting us.

I have enquired with the lab which tested this anti-CCR5 antibody in various applications, and they confirmed that it is not suitable for IHC, ICC/IF and IP, as also stated on the datasheet. For this reason (and because we cannot issue a testing discount code after purchase of a product), I am afraid I cannot be of help in this regard.

If she requires an anti-CCR5 antibody suitable for IHC-P on human tissue sections, we could recommend for example the mouse monoclonal ab110103 or the rabbit polyclonal ab65850. Should further specifications be required, I would be pleased to assist in finding the most suitable antibody.

Click here (or use the following: http://www.abcam.com/index.html?datasheet=110103).

Click here (or use the following: http://www.abcam.com/index.html?datasheet=65850).

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

Dear Sir/ Madam,
Sorry to disturb you. Our customer raises an enquiry about ab32048: Anti-CCR5 antibody [E164]. She has used ab32048 to conduct WB and the results confuse her. Her sample is mouse brain (cortex & striatum) and the dilution factor of ab32048 is 1/1000. There are signals both around 60 and 41 kDa (It is indicated in online datasheet that the predicted size of CCR5 protein is 41 kDa but observed band size is 60 kDa multiple bands were detected and the signal in 60 kDa was much strong than in 41 kDa.) She would like to know which band is the targeted signal. I am glad to provide the image if you need.  Would you please help this customer resolve this problem? Thank you very much.
Best regards,

ANSWER:

Thank you for contacting us.


The anti-CCR5 antibody [E164] (ab32048) when tested in house detected a band of ˜60 kDa in Jurkat, K562 and Daudicell lysate. This is most likelydifferent to the SwissProt referenced 41 kDa as the protein goes through post translational modifications (SwissProt: P51682). The following reference has reported that the 42 kDa form is mainly found in thecytoplasm whilst the 62 kDa form was mainly found in the cell membrane. Bothband of the gel maytherefore be of the CCR5 protein.


http://www.sciencedirect.com/science/article/pii/S0006291X02009087


They also observed that the 62 kDa form was more readily detectable under non-reducing conditions. Depending on how you are currently performing your western blot you may therefore be able to increase or decrease the band of62 kDa band by decreasing or increasing respectively the amount of reducing agent used.


I hope this information has been of help. If you have any further questions please do not hesitate to contact us again.