Anti-Parkin antibody (ab6177)

I appreciate very much your technical advise and great efforts on resolving the problems re Parkin antibody. The attached files are blots probing with different lot of the Parkin antibody (72864 and ACA0) within recent weeks. I hope the data can convince you that the quality of parkin antibody is not as good as before.

As i am in a hurry to get some decent blots for our revised paper (JNC), I sincerely hope you can provide me with the reliable quality of parkin antibody. If it is not possible, it will be understanding and i would not hesitate to seek from other suppliers.

Many thanks for your help.

ANSWER:

Thank you for sending me the images of your blots. Yes, there is a difference between the two lots that you have used. I'm sending you a replacement batch free of charge. For your record it is order number 65268 and it being shipped to your attention/JY932095. You should receive this by Friday. Please let me know how the replacement works out for you and if you have any additional questions.

BATCH NUMBER 72864 ORDER NUMBER -- NOT SPECIFIED --

DESCRIPTION OF THE PROBLEM High background Very poor signals

SAMPLE cell extract

PRIMARY ANTIBODY AB 6177, rabbit, 1:3000, RT * 1 hr Wash with PBS-T 6*10 min

SECONDARY ANTIBODY HRP-conjugated anti-rabbit AB, 1:2000, RT * 1hr Wash with PBS-T 6*10 min

DETECTION METHOD ECL-PLUS

ANTIBODY STORAGE CONDITIONS -20

SAMPLE PREPARATION RIPA+ PIC, 95 degree * 10 min

AMOUNT OF PROTEIN LOADED 20 ug

ELECTROPHORESIS/GEL CONDITIONS reducing 10-20% linear gradient SDS-PAGE gel

TRANSFER AND BLOCKING CONDITIONS Semi-dry transfer 5% dried-fat milk at RT * 1 hr

HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

WHAT STEPS HAVE YOU ALTERED? different lot of parkin antibody

ADDITIONAL NOTES Probbing with Parkin Antibody (lot number: ACA0) gave me clean and clear results. Unfortunately, using lot number 72864 antibody gave extremely high background and poor signal. I was wondering if you could replace me with the previous batch of antibody or another one? Thanks for your help.

P.S I purchased through King's College London in late-November. I will try to send you purchase number of King's when they return to work.

ANSWER:

Thank you for your enquiry and I'm sorry to hear that you are experiencing difficulty with ab6177. Can you be more specific about the cell extract you are using (eg - from which species)? Are your conditions the same as when you used the previous batch of ab6177? Did you try blocking with BSA to try to decrease the background you are seeing? Are you seeing any bands at all on your blot?

I can send you a newer batch of ab6177 to try, please indicate the purchase order number that was used. Thank you, and happy new year.

Is monoclonal antibodies against mouse parkin available with your company?

ANSWER:

We do not currently stock a monoclonal to mouse Parkin. The polyclonal antibody, ab6177, is all we have to offer against Parkin at the moment and this has not been tested in mouse. Many apologies.