Anti-C4d 抗体 (ab36075)

Thank you for the reply.

Could you please confirm whether either of the vials of ab36075 purchased showed any staining (one of the 20xxxx and the one from the xxxxx)?

Also, I asked for some feedback of how the antibodies had been stored. Has the customer provided this information?

I look forward to receiving your reply.

Thank you for contacting us. The antibody is supplied in liquid form and does not have to be reconstituted. The vial should contain 50 ul (0.2 mg/ml), if you do not see it you should spin it down. I can direct you to a starting dilution if you can tell me what application you plan to use.

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Thank you for your reply.

Having reviewed this case it seems that using the protocol the customer has applied the antibody should be producing staining.

May I ask, how has the antibody been stored? At what temperature and was it aliquotted?

The order details you gave to xxxx refers to an order of the 24th of September 2010. Was the antibody used before the initial complaint was made in April of this year? What were the results of these experiments?

Many thanks for your cooperation, I look forward to receiving your reply.

Thank you very much for having followed up with the customer and provided this additional information.

When looking through the protocol, I was surprised about the information on the secondary antibody. The customer mentioned to have used a rabbit anti human secondary antibody. This would not give a staining, as the primary antibody is a rabbit antibody. For a secondary antibody to detect the ab36075 an anti rabbit (e.g. goat anti rabbit, or donkey anti rabbit, ..) must be used. Can you please confirm this?

If the customer obtains no signal with an anti rabbit secondary antibody, we will be pleased to replace or credit this antibody for them. I apologize for the inconvenience.

For future experiments, I can also recommend strongly to the customer to include a blocking step. Indeed, blocking is a very crucial step for obtaining good images. Most of the time, 10% serum block is appropriate. As serum, we recommend to use serum of the species in which the secondary antibody is made (if a goat anti rabbit is used, we recommend therefore to use a goat serum to block). For further details or tips, please recommend the customer also our online protocols. https://www.abcam.com/index.html?pageconfig=resource&rid=11384

Thank you very much for your cooperation. I look forward to hear back from you.

We have previously been in contact regarding ab36075, which was not providing satisfactory results.

A free of charge replacement vial was issued and I am writing to inquire if the replacement product has worked. I would appreciate if you could let me know whether satisfactory results were obtained with this new vial as I am keen to know how you are progressing.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, please do not hesitate to let us know.

I wish you the best of luck with your research and please do not hesitate to contact us if you have any further questions.

Thank you for contacting us.

Your credit note ID is XXXXXXX.

I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

Thank you for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

Having reviewed the protocol details, I believe this product should have given satisfactory results. The only thing I can suggest now is manual staining with Microwave antigen retrieval method.

I apologize for the inconvenience and am pleased to offer you a credit note, or refund in compensation.

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

Thank you for this additional information.

I would be pleased to investigate this issue further to see whether we could provide some protocol tips. Can you please therefore send us the protocol details with help of the filled questionnaire (it should not take more than 5 minutes to fill) as well as an image of the result? This will help us to see whether there are any modifications which could significantly improve the results. If we are unable to improve the results, we will replace the antibody for the customer. Can you actually double check the order number you provided? I found there was a more recent order last year as well.

Thank you for your cooperation. I am looking forward to hear back from you.

Thank you for your response.

I am sorry to hear that you have been experiencing problems when using our products. Our anti-Twist antibody in guaranteed to work in mouse tissues in IHC-Fr. We have added recommended dilutions or specific protocols for this producthowever there are customer’s reviews in our Abreview section which do give some protocol information that these customers have used. I noticed differences between these Abreview protocols and was wondering if you could provide me with the protocol steps which you have used, any troubleshooting step that you taken and any images should you have these. If we cannot solve the issues which you are experiencing, I will be glad to send you a free of charge replacement, credit note or refund under our Abpromise guarantee.

We currently do not recommend using ab36075 anti-C4d on mouse tissues. The homology for the immunogen used to create this product and the mouse protein is below the threshold for cross reactivity. As such, I am very happy to look over your protocol and provide tips and suggestions which may help. However this product would not be covered by our Abpromise.

You are doing mouse on mouse staining with these products.Staining of mouse tissue using mouse antibody is a complicated process as high levels of background are often observed. It is notoriously difficult to eliminate this background.I would strongly recommend these tips for eliminating background when staining mouse tissue with a mouse monoclonal.

-1. Block endogenous IgG withunconjugated Fab fragment Anti-Mouse IgG (H+L) for 1 hr at room temperature, or overnight 4°C

-2. Block endogenous Fc receptors withF (ab) monomeric secondary antibodies

-3. Incubate sections with 1% Triton (in PBS) at room temperature for 30 min to ‘clean’ the tissue.

-4. Use TBS –Tween 20 as a washing buffer. This often gives less background than using PBS-Tween 20.

Further information may be found at:https://www.abcam.com/index.html?pageconfig=resource&rid=11465



Alternately I would suggest using theMouse on Mouse (M.O.M) Polymer IHC Kit, ab127055 which is designed toallow the use ofmouse antibodies on mouse tissueto obtain excellent signal intensities and clean background.(https://www.abcam.com/Mouse-on-Mouse-MOM-Polymer-IHC-Kit-ab127055.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.