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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. |
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#1
06-30-2008, 02:22 PM
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 GAPDH Not visible on western blot Hi, I am trying to detect GAPDH in prostate tissue and its not visible. Tried a variety of Ab concs and also different blockers but still no joy. I'm not sure if there are large amounts of GAPDH protein expressed in prostate tissue, does anyone know if there is and also what other loading controls are good to use? I'm going to try alpha tubulin but any other ideas would be good. Thanks!  |
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#2
07-02-2008, 02:48 PM
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| Re: GAPDH Not visible on western blot Have you been able to detect any other bands on Westerns using the same samples? What are your antibody concentrations/dilutions and the substrate being used? |
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#3
07-03-2008, 03:13 AM
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| Re: GAPDH Not visible on western blot Assuming that prostate tissue has no detectable levels of GAPDH, maybe try actin? |
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#4
10-02-2008, 03:13 PM
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| Re: GAPDH Not visible on western blot ethidiumbromide has the right idea. I have found that Beta-actin is very reliable. I use it at about a 1:10,000 concentration. I have also been having troubles with my GAPDH as of late. After searching the web, I have learned that GAPDH will not reliably band when blocked with anything other than a 5% BSA. Hope that helped a bit, Good luck! |
| The Following User Says Thank You to anna.oskar For This Useful Post: | ||
mbsum09 (07-17-2009)
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| blot , gapdh , visible , western |
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