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Anti-RACK1 antibody (ab27982)

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1 question for ab27982

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Question 1

Tuesday 16-May-2006

BATCH NUMBER 182066 ORDER NUMBER 104789

DESCRIPTION OF THE PROBLEM No signal.

SAMPLE HEK cell lysate

PRIMARY ANTIBODY this rack ab at concentrations of 1, 2 and 4 ug/ml, RT for 3hrs

washed 4 times 5 min tbst

DETECTION METHOD ECL

POSITIVE AND NEGATIVE CONTROLS USED i also used another RACK1 Ab under the same conditions and saw bands after 30sec film exposure to blot.

ANTIBODY STORAGE CONDITIONS aliquotted and stored at -20 frosty freezer

SAMPLE PREPARATION lysed cells in SDS/Beta-ME containing sample buffer

ELECTROPHORESIS/GEL CONDITIONS 12% reducing

TRANSFER AND BLOCKING CONDITIONS 5%milk in TBS + 0.1% tween blocked for 1 hr rt shaking

SECONDARY ANTIBODY anti-rabbit 1:10000 RT 1hr

washed 4 times 5 min tbst

HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

WHAT STEPS HAVE YOU ALTERED? I tried an O/N 4degree incubation with primary. I cant think of any other steps as these conditions work with my other RACK1 Ab and therefore I know all the reagents are good.

ANSWER:

  

Thank you for your enquiry.

I am sorry to hear that you have been having difficulties with this antibody. You are using an approach that I would largely recommend. You are clearly presenting the antigen on the membrane as shown by the detection of this target using another antibody.

I am certainly prepared to offer you a credit note were the antibody purchased within the past 90 days. If this is the case please provide me with your order details and date of purchase and I will arrange for our accounts department to raise credit.

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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