Anti-CCR2 antibody [E68] (ab32144)

Customer kindly called to discuss a problem which has arisen with their use of anti-CCR2 E86 in flow cytometry. While this does give an excellent peak in their normal flow experiments, recent experiments using a CCR knock out line has yielded the same peaks. The KO line has been sequenced and is know tho not express. Their conversations with Jackson Labs have yielded the possibility that this has occurred in previous similar experiments before. They ask if we have any knowledge of this type of issue and if we could please provide the reasoning/data that lead to our statement " The antibody does not cross react with other CKR family members."

ANSWER:

Thank you for contacting us regarding the issues which you have faced using ab32144.

The lab is very interested in the discussions which you had with Jackson regarding this spike in the knock out mouse. Could you please provide any additional details regarding this? Was this with another Abcam product or that of another group? Was Jackson speaking about this mouse line in particular?

Any details that you could provide including but not limited to this would be of great help in trying to resolve this problem for you.

I look forward to your reply. Please let me know if you have any questions.

Can we get a refund as originally requested instead of credit? Or can we choose to use this credit to receive a refund? Also for ab32144 (credit note ******).
Thank you and have a good weekend,

ANSWER:

Thank you for your reply.

I have forwarded you enquiry to our accounts team (mailto:us.creditnotes@abcam.com) and they will look into this and get back you .

Please let me know if there is anything else I can help you with.

Hope you have a great weekend.

Dear Abcam-Team,
could you please check the concentration of the ab32144 Lot: GR1-14954-1,
since I need it for the isotype control for flow cytometry analysis.
Sincerely

ANSWER:

Thank you for your inquiry.

I am happy to confirm that the concentration of lot 993363 (GR14954-1) is 0.579mg/mL.

I also would like to comment on the seemingly low concentration:

The IgG produced by rabbits have greater affinity and greater sensitivity than mouse IgG (about 100-1000 fold) and at the same time the total amount of IgG produced by the rabbit hybridomas is generally lower than in mouse hybridomas.

Many 'veteran' antibody users are generally familiar with the IgG amount provided from mouse antibodies and can judge how many reactions they may get or need to use based on concentration. Unfortunately, this experience is not transferable to rabbit monoclonal antibodies because the concentration scales are generally very different.

Since the specificity and sensitivity is much higher with rabbit monoclonal antibodies, less IgG is required per reaction.

I would like to reassure you that while the amount of IgG may not be comparable to mouse monoclonal antibodies, customers are receiving a comparable number of reactions/tests/blots/stainings per tube.

I hope this information is helpful. Please do not hesitate to contact me with any further questions in this matter.

I would like to work on CCL2/CCR2 signalling pathway in sheep and as such I am looking for an anti-CCR2 antibody that will cross-react with sheep. I have not found any except E68 that cross-reacts with 4 other spp but has not been tested in sheep. Before I decide to purchase this, can I get a sample to test in our lab? Let me know if this would be possible.Thanks in advance.

ANSWER:

Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

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 intersted in participating in this program and testing ab32144 in sheep please let me know. Also, please do not hesitate to contact us with any other questions.

WHy was the ICC/IF application removed from this antibody. What cell types were giving non-specific staining?

ANSWER:

Thank you for your inquiry, and I deeply apologize for the delayed response.
Unfortunately, because our retest result showed negative, we do not have the image which can be sent to you. We do not have any specific information as to which cells were staining non-specifically; however, the result showed a weak and non-specific staining - nuclear staining was observed.
I am sorry that we are not able to provide you with any further details.
Please let me know if there is anything else I can help with.