Anti-Histidine decarboxylase antibody (ab37291)

Getting signal from human samples in westrn blot, but not from mouse or rat.

ANSWER:

Thank you for calling Abcam earlier today.

I am sorry that you were having some problems using ab37291 in western blot. As we discussed on the telephone, I am sending you a new vial of this this to try out. Your new order number is ********.

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

I got this product from a distributor. So if you agree to change another antibody, I will ask them for the oder number.

ANSWER:

Thank you for your reply.

I am happy to offer you a replacement product. I can arrange this through the distributor - please provide the distributor name. Also please indicate if you are okay to receive another vial of this antibody or a credit.

I look forward to your reply so that I may process your request.

I am having trouble with this antibody in WB. I purchased it at october, if possible, I want a replacement, Thank you!

ANSWER:

Thank you for your reply.

In order to send a replacement I will need a specific order number. Did you order the item directly from Abcam or through a distributor?

I look forward to your reply so that I may assist you further.

Thank you for your suggestions. As you know, I have problems with three antibodies, the problem of two is have no apparent band(ab3611 and ab15323), and the other is with multiple bands(ab37291), would you like to give me a total suggestions for these antibodies？ Thank you!

ANSWER:

Thank you for your reply.

I can see that my colleagues are assisting you with ab3611 and ab15323. The protocol suggestions that I have given you are specific for ab37291 and I see that they have also offered protocol suggestions for these products.

Regarding ab37291, I would recommend decreasing the antibody concentration as we have found this to work well down to 0.5ug/ml. If this does not improve your results, please let me know and I am happy to offer a replacement or credit.

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

DESCRIPTION OF THE PROBLEM Multiple bands SAMPLE rat tissue PRIMARY ANTIBODY 1:100 in 5%milk in TBST 4℃overnight DETECTION METHOD ECL plus POSITIVE AND NEGATIVE CONTROLS USED no ANTIBODY STORAGE CONDITIONS -20℃ SAMPLE PREPARATION RIPA，pmsf， protease inhibitors（rotch）lysis buffer； 100℃5min to denaturation AMOUNT OF PROTEIN LOADED 50ug ELECTROPHORESIS/GEL CONDITIONS 8-10%gel 80v,30min 100v,1hour TRANSFER AND BLOCKING CONDITIONS wet transfer, 100mA,100-120minutes blocking: 5% milk in TBST two hours SECONDARY ANTIBODY 1:5000.5%milk in TBST 2 hours at room temperature HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? add the wash times

ANSWER:

Thank you for contacting Abcam regarding ab37291.

I am sorry that you have been experiencing difficulties with this antibody in WB. I have reviewed the protocol information you provided and I would like to make some suggestions to improve your results.

Regarding the concentration of primary antibody being used, 2ug/ml is quite high. I would recommend decreasing the concentration to 1 or 0.5ug/ml which should reduce the background you observe. It may also be necessary to reduce the incubation time to 1-2 hours at room temperature rather than overnight.

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