Quick/One-step/automated Western Blot
Posted 23 November 2011 - 05:05 AM
I've seen companies promote a "one-hour" detection system etc (http://www.genscript...stern_tech.html) but is there something like a PCR machine out there yet?
Posted 24 November 2011 - 12:47 AM
Posted 24 November 2011 - 01:03 AM
One day WB = 6 AM to 6 PM?
Total protein extraction (15 min - 2 h depending on how many samples you have), total protein quantification using BCA method (30 min), SDS-PAGE (3-4 h), transfer (2-3 h), blocking (1 h), primary (1h), secondary (30 min), total washing (30 min), visualise (15 min). Sounds plausible to me. Problem is I deal with large sample sizes
Posted 24 November 2011 - 01:12 AM
I generally run 8-12% gels which take >2 h just to get the dye front off the gel @ 100V. I'm fearful of overheating if I go beyond that.
And do semi-wet transfers take shorter time vs wet transfer? I always do wet transfers.
Posted 24 November 2011 - 01:16 AM
Posted 24 November 2011 - 01:22 AM
I should probably try out your running/transfer settings. it could really save alot of time albeit being in the lab the whole day.
Do you find that an overnight transfer gives better results vs. hourly transfers?
And, overnight primary antibody incubation at 4 degrees C vs. 1 h RT incubations?
Posted 24 November 2011 - 01:25 AM
Posted 24 November 2011 - 01:29 AM