Anti-PERK 抗体 (ab65142)

Thank you for the clarification.

Have you considered using an antibody concentration kit, like our ab102776, for a buffer exchange? This may be more cost-effective than having the lab do a custom preparation.

https://www.abcam.com/index.html?pageconfig=resource&rid=13489

https://www.abcam.com/Antibody-Concentration-Kit-1-column-ab102776.html

If you are strictly interested in the ab65142 prepared without azide, I will double check with the lab to see if this is possible. Let me know what you think.

Thank you for your inquiry.

The formulation for ab65142 is:

Preservative: 0.02% Sodium Azide
Constituents: 50% Glycerol, PBS (without Mg2+ and Ca2+), 150mM Sodium chloride, pH 7.4

It does not contain any carrier proteins like BSA. Are you looking for an azide-free or glycerol-free preparation? I just want to make sure before I contact the lab. I look forward to your reply.

Thank you for contacting us.

Your credit note ID is XXXXXX.

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:

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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 very much for your email.

We unfortunately do not have any more suggestions to make. We are not sure about the reason of non specific band so in order to understand more about the problem I am happy in providing a free of charge replacement vial.

Could you confirm how you would like to proceed?

Thank you for your enquiry regarding ab65142 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. Additionally, thank you for supplying an image, as this has been very beneficial in understanding your concerns.

The protocol looks fine to me however I would like to make few comments/suggestions that might help to improve your results and that you could consider trying:

- Use ice cold RIPA buffer and try fresh protease inhibitors
- Use 70-80% confluent cells for preparation of lysates
- We always heat lysates in sample buffer for 10 minutes at 100C
- We always use 5% BSA for blocking for 2 hours
- In our lab we wash the membrane once for 10 minutes and 3 times for 5 min with TBST
- Try no primary control

I hope these suggestions will help to improve your results. If in any case the results do not improve please do not hesitate to contact me for further assistance.