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Anti-Ran antibody (ab4781)

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1 question for ab4781

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Question 1

Tuesday 14-November-2006

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED --

DESCRIPTION OF THE PROBLEM No signal or weak signal. NO sSIGNAL.

SAMPLE I have generated a recombinant Leishmania Ran with very high homology to the human ran (-80%). I have loaded 2ug of purified protein and there was no signal at all even at a dilution of 1:150. Is this normal for a polyclonal antibody? Is the antigen a small peptitde or the whole protein? On the contrary when I do immunofluorescence there seems to be nuclear staining in Leishmania samples. Please help!!

PRIMARY ANTIBODY 1:500, 1:150

DETECTION METHOD DAB peroxidation

SAMPLE PREPARATION Recombinant protein.

TRANSFER AND BLOCKING CONDITIONS 5% milk in TBS and O/N incubation 4 oC.

SECONDARY ANTIBODY [another company] anti-rabbit HRP 1:2000

HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes

ANSWER:

  

Thank you for your enquiry. This particular antibody has not been tested in the species that you are looking at, so we can not guarantee that the antibody will work. Since the antibody is working for immunofluorescence, I would think that there could be something wrong with your Western blot protocol. Have you tried using the specific Western blot protocol listed on the antibody's data sheet? I have given you a link to the data sheet below, the specific Western blot protocol can be found under the protocols tab.

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

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