Anti-Transferrin Receptor antibody [MEM-189] (ab1086)

I am a interested in purchasing a primary anti-transferrin receptor antibody (ab1086) which, according to the datasheet, reacts with human, mouse and rat. However, because this antibody is raised in mouse I am worried it will react to more than my protein of interest when using mouse samples. This is an issue that I ran into in the past and I had to introduce an additional blocking step to eliminate non-specific binding of such primary antibody with mouse proteins. Is this something that I should take into account with this antibody as well? and if so what kind of blocking do I have to do?
Many thanks in advance
Best regards

ANSWER:

Thank you for contacting us.

Yes, the extra blocking step is always required to avoid the mouse back ground. We have recommendations that can be found by clicking the following link;

http://www.abcam.com/index.html?pageconfig=resource&rid=11465

We have kit which is perfect for mouse on mouse staining; please have a look at the datasheet

http://www.abcam.com/Mouse-on-Mouse-MOM-Polymer-IHC-Kit-ab127055.html

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

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Would these products
(ab1086, ab84036, ab9179, ab8598) detect Trf1 and Trf2?

ANSWER:

Thank you for your enquiry.

We have not tested these products (ab1086, ab84036, ab9179, ab8598) for cross reactivity with Trf2. However, based on immunogen sequence (ClustalW analysis, Score 23.0), we do not expect to see any cross reactivity whatsoever.

I hope this information will help settle any concerns that you may have.

Thank you for your help. I have stored them as the product information indicated, long term in the -20. But I understand keeping the freeze thaws down. I am running some more westerns soon and will include TFRC ab1086 in the bunch. I will keep you posted on the results. If the bands come out at the wrong molecular weight I will be able to get a refund, but not actually return the antibodies? Also, out of curiosity, why is this particular antibody to be used on non reduced samples? Thanks again,

ANSWER:

Thank you for bringing that to my attention.
I did some additional research into CD71. CD71, also known as the transferrin receptor (TFR), is a type II membrane glycoprotein that exists as a disulfide-linked homodimer of two identical subunits. Ab1086 used under non-reducing conditions should recognize the homodimer at greater than 170kDa after post translation modifications. Did you see any bands in that range?
If used in reducing conditions it seems that the product should pick up the band around 85kDa (before PTM). The datasheet is not very clear about this. When repeating your experiments I would recommend looking for the larger weight. It may also be useful to try reducing conditions as well.
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.

Great to hear I can return them. They have been stored in the -20 since arrival. I am not sure what information you need about the original vial so I will give you what I have and see if you have more questions. I lysed the cells in SDS free RIPA buffer with protease inhibitor tablets EDTA free from Roche. Centrifuge to remove DNA. BCA for protein concentration. 10ug protein with 4x loading dye invitrogen, no beta mercaptoethanol. Run on a nupage gel in MOPS SDS running buffer. Transfer to a nitrocellulose blot. Block in Licor brand blocking buffer, use ab1086 at 1:800 over night in blocking buffer and beta actin at 1:5000. Wash in TBST. Incubate in licor anti mouse and rabbit 1 hr. wash in TBST, read on scanner.

I will look into getting the order number for the original order, but that may take a little time. I will email when I have that information. I do know that it arrived on if that could narrow the search for you. Let me know if there is any other information you need.

ANSWER:

Thank you for your response.

I am very sorry to say this but we cannot accept returned items that have been frozen. This is because we need to limit any freeze/thaw cycles that the product might experience. I am sorry about the inconvenience that this might cause.

May I suggest however that you repeat your experiment using one of these vials? I have had the opportunity to review your protocol, which looks fine. If your results are the same (a band at less than 1/2 the expected weight) then we might conclude that the issue is with the product. In which case I might refund or replace all the products without need for return.

Please let me know if this is acceptable for you and if you have any questions.

I recently started using the Abcam antibody AB1086 for TFRC.  I ordered one tube and used it and got a positive signal on a western blot using the non reducing conditions indicated on the data sheet provided.  I recently ordered more to use in a FACS experiment as a control.  I have now realized that the band I was taking as positive signal is not of the proper size.  TFRC is 84kD and the band I see is at about 45. I would like to return the three new tubes I bought if possible as this antibody is not suitable for my experiment. . I am attaching a PPT slide with the western image. Please let me know if you need any more information and how you would like to precede. material from any computer.  Thank you.

ANSWER:

I am sorry that you have been experiencing problems with one of our products and wish to thank you for bringing this to our attention. As this order was placed and delivered within the last seven day I may issue a return for you. Since this is ultimately a problem with one of our product I am waiving the standard 20% restocking fee for you.

Before I may process this return however I will need a little information:

Your order was for three vials of ab1086. Have any of these been opened?

How have these products been stored since delivery?

I would also like to discuss the original vial of ab1086 with you and perhaps issue a free of charge replacement or refund for that product. In order to do this I will need some information about your experiments with this product. Could you please send me the protocol used?

Would you also be able to send me the order number for the original vial?

Please let me know if you have any questions.