Anti-CCR5 antibody (ab7346)

Good day   I am a postdoctoral scientist at the NIH and I am interested in purchasing one of your CCR5 peptides. However I am not sure which of the peptide products (ab92719, ab95861 or ab7876) might be more suited to my experiment.   I am working on anti-HIV targets and I want to test (via ELISAs) if my target inhibits the gp120-CCR5 binding. Could you recommend one of your CCR5 peptides (and corresponding anti-CCR5 antibody) that might be most suited for my application?   Thanks and regards

ANSWER:

Thank you for contacting Abcam.



These peptides are very short fragments of the CCR5 protein and as such may not contain the gp120 binding site. You may wish to use the whole protein. While this is not currently available at Abcam, you may find this by searching biocompare.com These peptides (ab7876ab92719 and ab95861) are the immunizing antigens for antibodies ab7346, ab64620 and ab85837 respectively.



None of the antibodies has yet been tested in ELISA. For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:

The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: http://www.abcam.com/collaborationdiscount.



If you would be interested in this offer please let me know.



The product pages for these antibodies are at:

http://www.abcam.com/CCR5-antibody-ab7346.html



http://www.abcam.com/CCR5-antibody-mAbcam-64620-ab64620.html



http://www.abcam.com/CCR5-antibody-ab85837.html



I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

Could you tell me if the epitope recognized by your CCR5 (ab7346) corresponding to amino acids 6-20 of Human CCR5 is also present in HIV Clade C population, please?

ANSWER:

Thank you for contacting us with your question.

I aligned the immunogen sequence SSPIYDINYYTSEPC against the HIV M:C (taxid 505186) protein database and no putative conserved domains were identified.

I hope this information is helpful, but please let me know if you have further questions.

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED --

DESCRIPTION OF THE PROBLEM No CCR5 band, only background bands.

SAMPLE Human sample from blood derived Mature Dendritic cells. Purified protein using Pierce M-PER reagent.

PRIMARY ANTIBODY Primary Antibody: Abcam rabbit polyclonal CCR5 at 1:1000, diluted with 3:1 with TBS-T and NAP-Blocker and incubated at 4C overnight. Wash 30 minutes with TBS-T

Attempted second time at advice of abcam tech, using 5:1 dilution of TBS-T and NAP-blocker. Wash 30 minutes with TBS-T

DETECTION METHOD Detected with 45 minute step of Invitrogen Streptavadin 1:2500, then 30 minute wash. Developed with Pierce NBT-BCIP solution at RT until bands develop.

POSITIVE AND NEGATIVE CONTROLS USED None

ANTIBODY STORAGE CONDITIONS Stored at +4C

SAMPLE PREPARATION Used 1:1 Laemmli Sample Buffer that has 5% B-MERC. Heated sample for 5 minutes at 95C.

AMOUNT OF PROTEIN LOADED 30 micrograms per lane

ELECTROPHORESIS/GEL CONDITIONS GeneMate Express Polyacrylamide Mini-gels. 4-20% Express gels

TRANSFER AND BLOCKING CONDITIONS Transfer: Transfered at 25V for 100 minutes onto PVDF membrane. Blocker: G-Biosciences NAP-Blocker 1:1 with TBS-Tween. Blocked for 30 minutes at RT.

SECONDARY ANTIBODY [another company] goat anti-rabbit, diluted 1:2000 in 3:1 TBS-T and NAP-blocker. Incubated at RT for 2 hours. Wash 30 minutes with TBS-T

Attempted second time using 5:1 dilution of TBS-T and NAP-blocker. Wash 30 minutes with TBS-T

HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

WHAT STEPS HAVE YOU ALTERED? I have changed the amount of blocking reagent in the Primary, secondary, and Streptavadin solutions from 3:1 TBS-T/NAP-Blocker to 5:1 TBS-T/NAP-Blocker with no different results.

ANSWER:

Thank you for your enquiry.

I am sorry to hear that you have been having difficulties with this antibody. I have read your technical questionnaire and I appreciate the additional steps that you have performed in order to try and get this antibody to work.

I am certainly prepared to offer you a credit note against this purchase provided that the antibody was purchased within the past 90 days. If this is the case please email me details of the order including the order number and date of purchase. I look forward to hearing from you.

This email is additional information for my previous email whichi subject is "Question about antibody and peptid".

I forgot to ask you how pure they are and which method these peptides have been purified by.

Thank you for your cooperation.

ab2001, HREALEKDIKTSYIMDHC ab7346, SSPIYDINYYTSEPC ab3430, MSDFDSNPFADPDLNNPC ab3346, MSAPAQPPAEGTEGAAPGC ab3347, MSATAATVPPAAPAGEGGC

Thank you for your cooperation

ANSWER:

Thank you again for your enquiry. Regarding ab4969, ab4957, and ab4958 (the peptides corresponding to the antibodies you indicated), they are all greater than 95% pure, and they were purified via HPLC.

If you have any further questions, please contact us again.

This email is additional information for my previous email whichi subject is "Question about antibody and peptid".

I forgot to ask you how pure they are and which method these peptides have been purified by.

Thank you for your cooperation.

ab2001, HREALEKDIKTSYIMDHC ab7346, SSPIYDINYYTSEPC ab3430, MSDFDSNPFADPDLNNPC ab3346, MSAPAQPPAEGTEGAAPGC ab3347, MSATAATVPPAAPAGEGGC

Thank you for your cooperation

ANSWER:

Thank you for your enquiry. For ab2001 and ab7346, the peptides are non-HPLC, which averages out to 60-70% purity. Ab2001 (the antibody) was immunoaffinity chromatography purified IgG, and ab7346 (CCR5 antibody) is purified IgG.

I will have to get back to you regarding the other peptides.

If you have any additional questions, please let us know.