Anti-GPCR TGR5抗体(ab72608)

I am sorry to hear that you have been experiencing problems using this product in the application that you wish.

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Could you please provide some further details of the protocol used and complete the following form (attached as a word document).

Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

We expect that glycosylation will apeear as a smear on the blot, rather than distinct bands, so I am skeptical that you are detecting TGR5. In addition, you should be seeing more signal in the membrane fraction, and not an identical banding pattern in the cytosolic fraction.

Can you please tell me how the samples are prepared before loading into the gel? That is, how are the fractions prepared, and how are they denatured before loading into the gel? In general, for multi-pass membrane proteins, our recommendation is the note on the datasheet:

GPCR is multi pass membrane protein, customers may try heating the lysates for 15-20 minutes at 60-70C instead of boiling the samples. Non fat dry milk as blocking agent seems effective than BSA with current lot.

L’anticorps ab72608 n’a pas été purifié et est un antisérum entier. La concentration n’a donc pas été déterminé par contre nous estimons que la concentration sera de 1-10 mg/ml.

Thank you for your enquiry regarding ab72608 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

The protocol looks absolutely fine to me, I would only suggest loading less lysate ie. 15-20 ug per lane and running a positive control (human, mouse and rat spleen lysates) alongside if you have access to this type of tissue.

According to Swiss-Prot website, this protein can be highly N-glycosylated. There are 3 potential site alongside the polypeptide chain at 5, 75 and 153 aa. This could result multiple band ob the blot. Please take a look at the following site for further information:

http://www.uniprot.org/uniprot/Q80T02

If you need any further assistance in the future, please do not hesitate to contact me.



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Thank you for contacting Abcam about ab72608.

We are unable to provide the peptide that was used to generate ab72608 (it was all used up for development and testing of the antibody). However, you should be able to have it readily synthesized for a low cost at a facility local to you.

Here is the immunogen sequence:
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Please let me know if there is anything else I can help you with.

Thank you for your email.

I am sorry to hear that the antibody did not work. I can help you with troubleshooting so that you only have to repeat the experiment once; although there was a typos error in image but it should not have affected the specificity and quality of this antibody.

Could you please fill the attached questionnaire?

Many thanks for your cooperation in this case. I will look forward to hearing from you soon.

Thank you for your enquiry.

I can confirm that ab72608 antibody is sold as whole antiserum. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datasheet. Antibody concentration is usually determined by protein assay, and serum / ascites / tissue culture supernatant will contain a lot of other proteins, which means the antibody quantification would not be accurate.

I can confirm that for whole antiserum, concentration of antibody is known to very between 1 - 10 mg/ml.

I am sorry we are not able to provide an exact concentration on this occasion, but hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

PS: We apologies for the error on datasheet which has been corrected.

Thank you for your enquiry.

I can confirm that ab72608 antibody is sold as whole antiserum. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datasheet. Antibody concentration is usually determined by protein assay, and serum / ascites / tissue culture supernatant will contain a lot of other proteins, which means the antibody quantification would not be accurate.

I can confirm that for whole antiserum, concentration of antibody is known to vary between 1 - 10 mg/ml.

I am sorry we are not able to provide an exact concentration on this occasion, but hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Thank you for contacting Abcam.

I am sorry that you were experiencing some difficulties with ab72608 in western blot. If the protocol variations that we talked about do not prove to be successful, then please let me know and I will be happy to either replace or refund that antibody for you.

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

Thank you very much for your reply.

I will be happy to send a fresh vial to try. Could you let me know the PO or order number once your purchasing agent has found this information? I will need it to process the replacement.

Please let me know if you have any questions or if there is anything else that we can do for you.