Haynes Lab:Notebook/CRISPR Editing/2016/03/01: Difference between revisions
Rene M Davis (talk | contribs) |
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get a square dish to image the membrane in. remove one membrane from buffer, dab the edge with a kimwipe to remove extra liquid, place protein side up in dish<br> | get a square dish to image the membrane in. remove one membrane from buffer, dab the edge with a kimwipe to remove extra liquid, place protein side up in dish<br> | ||
image. was difficult to get a good pic because our background was really high. can try to fix by setting the exposure area to the highest signal. | image. was difficult to get a good pic because our background was really high. can try to fix by setting the exposure area to the highest signal. | ||
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'''conclusion:''' background signal was really high, likely because the blocking buffer was less stringent than when KAH did hers with milk. can compare this image to the ponceau stain, see if the ratio of the stained bands is higher. should redo with much less secondary antibody, maybe dilute 1:3000 or more. | '''conclusion:''' background signal was really high, likely because the blocking buffer was less stringent than when KAH did hers with milk. can compare this image to the ponceau stain, see if the ratio of the stained bands is higher. should redo with much less secondary antibody, maybe dilute 1:3000 or more. |
Revision as of 21:01, 12 March 2016
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03/01/2016Western secondary antibody and imaging
STAINING HRP
set the membrane protein side down so it soaks in the bubble. drag the membrane until the bottom edge is just touching the bubble, then slowly lay the membrane across. Transfect Luc14s
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