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Western Blot Forum Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


No Bands on Western Blot

No Bands on Western Blot - Western Blot Forum

No Bands on Western Blot - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


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Old 08-27-2007, 01:51 PM
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Default No Bands on Western Blot



Hi,I am new here.

I am getting no bands on my western

what should i do?

Last edited by admin; 08-27-2007 at 01:54 PM.
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Old 08-27-2007, 01:51 PM
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Default Re: air bubble while loading sample.

Ok will look into this.
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Old 08-27-2007, 04:26 PM
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Default Re: No Bands on Western Blot

1. Check transfer efficiency, stain used gel after transfer, or protein blot on membrane.
2. Check any reference material to see if you should be running denatured or native proteins.
3. If you are running protein mix, like a cell lysate, what percent or amount should be your target protein.
4. Troubleshoot the Antibody concentration by running the positive control in multiple lanes, blotting, cutting membrane into strips and blotting one condition on strip at a time until find best conditions.
5. Check your detection reagents.
6. Check your camera or scanner. (increase exposure time)
7. You can even try blotting by skipping the blocking step, but expect to see nonspecific binding at areas of high protein concentration though. You may see a very weak signal this way at your expected size range though.
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