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Dear Technical Team,
Hope this e-mail finds you well!
Please find below a complaint from one of our customer to whom we supplied
antibodies having cat nos. ab94585, ab11259, ab124025 & ab31928.
The technical forms are also attached with this mail.
Request you to kindly look into it & advise us accordingly to do further
needful on it.
Looking forward towards your valued reply/ comments on it.
Thanking you in advance.
With Best Regards,
Asked on May 22 2012
Thank you for taking the time to complete our questionnaires and contact us. I am sorry to hearthe customer has had difficulty obtaining satisfactory results from these antibodies.
The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.
To confirm with you, four antibodies are mentioned in your email. However, 2 copies of the questionnaire for ab124025 have been provided and there is no questionnaire for ab11259.
I can provide a response for ab94585,ab124025and ab31928. If you require further help regarding ab11529, please do not hesitate to send the questionnaire and I will be pleased to help.
I would like to reassure you that ab94585 and ab31928 are tested and covered by our 6 month guarantee for use in IHC-Fr and mouse samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement. Regarding ab124025, this is tested and guaranteed in mouse, but has not been tested in frozen sections. Could you confirm if the customer has tried paraffin sections, for which we can provide the guarantee?
Reviewing this case, I would like to offer some suggestions to help optimize the results from ab94585,ab124025and ab31928. I would also appreciate if you can confirm some further details. There are similar comments to provide for all three antibodies:
1. Fixation 1 hour for frozen sections is a very long time. For Frozen sections, we recommend fixation for 10 minutes only. Otherwize there may be too much protein crosslinking from the PFA, and the epitope may not be accessible to the antibody.
2. As far as I am aware, Recoverin, GABA A Receptor alpha 1 and Neurokinin 3 Receptor NK3 are all membrane or membraneassociated protein.Detection of thesewill not require permeabilization. Indeed, Triton is a strong detergent and can disrupt membrane proteins and eptiopes, which can significantly affect the staining. Therefore, I can suggest to remove the permeabilization step.
3. I can recommend to use a more gentle detergent, such as Tween 20 in the other buffers, try 0.1% - 0.2% Tween.
4. I recommend to include 0.1 - 0.2% tween in the wash buffer, this will help to wash away any excess antibody.
5. We recommend not to mix blocking agents in an experiment. try serum only.
6. Could you confirm, are the current vials of secondary antibodies working well with other primary antibodies? What were the results of the no primary controls? The concentration of secondary antibody may need to be reduced to help optimize the results.
I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details, including information regarding ab11259.
Answered on May 22 2012