Anti-FANCD2 antibody (ab2187)

I'm looking for an antibody that will recognize FancD2 in mouse. Your product sheet says that ab2187 should do this, but I don't see any references in which it has been used in mouse. Can you tell me the basis for your belief that it will cross-react?

ANSWER:

Thank you for contacting Abcam regarding ab2187.


I can confirm that this antibody gave positive results using IR treated MEF cells. Additionally, this antibody is covered by our Abpromise guarantee which states that we will replace or refund any product that does not perform as specified on the datasheet when reported within 6 months of purchase.


I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

Hi Kate,

Please see below the response from our customer;

Thanks for getting back so soon. I would like to get a replacement of AB2187 preferably with a different lot number. Thanks again for your help.

Thanks for your help!

Kind regards,

ANSWER:

Thank you for kindly confirming these details.

As requested, we would be pleased to arrange this free of charge replacement. In order to do this, I am forwarding these details on to our distributors team who will be happy to organize this for you.

Original order number: ####
1 X vial: ab2187 FANCD2 antibody
Original lot number:GR48772-3

Please provide a FOCR from a different lot number for this customer.

Thank you for your help and cooperation with this case. Please do not hesitate to contact us if you need anything further.

LOT NUMBER GR48772-3
ORDER NUMBER PO-12372
DESCRIPTION OF THE PROBLEM With previous antibody purchased (June, 2010) lot #833716, I got nuclear foci in mouse embryos. With the new antibody purchased (Nov, 2011), there is complete absence of staining in the nucleus. Steps and reagents used has all been the same.
SAMPLE Mouse Embryos (Grown in vitro) PRIMARY ANTIBODY Abcam, AB2187, in 1% BSA, 0.1% Tween at 1:500 for 2hours at room temp. wash 3x15 mins with diluent.
DETECTION METHOD Confocal Microscopy
POSITIVE AND NEGATIVE CONTROLS USED No primary, as well as different secondary.
ANTIBODY STORAGE CONDITIONS Immediate storage at -80C in single use aliquots
FIXATION OF SAMPLE 2% PFA with Triton X permeabilisation for half hour at room temp
PERMEABILIZATION STEP Triton X, together with fixing
BLOCKING CONDITIONS 3% Goat Serum in 1% BSA, 0.1% Tween –PBS for overnight in room temp
SECONDARY ANTIBODY Invitrogen Alexafluor 488 Anti-rabbit goat, in 1% BSA, 0.1% Tween at 1:1000 for 1hours at room temp. wash 3x15 mins with diluent
HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4
HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes
DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes
WHAT STEPS HAVE YOU ALTERED? Different secondary,
ADDITIONAL NOTES I have used the same method stated above with 2010 antibody to obtain nuclear foci. is a previous lot available as a replacement?

ANSWER:

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will provide us with vital information for our monitoring of product quality

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been succesful. Reviewing the details, I can confirm that this antibody has not been tested in whole mount, IHC-Fr or IHC-P and would not usually be covered by our 6 month guarantee for these untested applications. However, as the previous lot has worked in whole mount in this case, I am pleased to make an exception.This indicates there are no further majortips to provide on this occasion to help improve the results.

I apologise for the inconvenience and am pleased to offer you a free of charge replacement or credit note in compensation.

With regards to lot numbers, I am sorry to confirm that we don't have any of lot 833716 leftin stock. I would like to reassure you that we monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of this antibody or this newer batch. Regrettably, I can suggest you have received a bad vial on this occasion. Therefore, we would be pleased to provide a replacement from a different lot, which will still be covered by our guarantee. Alternatively, I fully understand your concerns and you prefer a credit note in this case, I will be pleased to arrange this for you.

In addition, I can suggest you may like to consider trying staining overnight 4oC.which may help to provide more consistent results between different lots. It will also allow a longer time for the antibody to penetrate through the larger peice of embryonic tissue.

There is further information on whole mount including troubleshooting tips on our webpages which may be helpful:

http://www.abcam.com/index.html?pageconfig=resource&rid=11330

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

The customer wished toknow whether we have any products to this target that will not cross react with GST fusion.

ANSWER:

Thank you for your enquiry.

In my colleague Charlotte's absence I am dealing with her enquiries.

I have heard back from the source of ab2187 and this antibody was raised using an N-terminal fragment GST-fusion protein. If you are interested in an antibody that does not demonstrate reactivity against GST I would like to recommend the three monoclonal antibodies that we have against this target. These will not show reactivity with GST given their single epitope specificity.

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Sorry, I have looked the datasheet carefully, but can not find the information about which fragment of D2 protein is used to raise this poyclonal Ab. Look, I CAN also get very nice IP results by monoclonal Ab offered by SantaCruz. This is the issue here. My problem, is in certain circumstance I want to study, the IP works badlly, which could be due to epitope masking. That is why I need to know which epitopes your Ab is against. I want some Ab recognizing a different region of D2 other than the N terminal, if that is a commercial secret (though I understand why such information shall be kept in closet), I wonder if I can get some free sample to test. Thank you sir, Best regards, Gang

ANSWER:

Thank you for your enquiry. Our FANCD2 polyclonal antibody has been made to a human FANCD2 fusion protein directed against the N-terminal portion of the protein.