No straight band-WESTERN BLOTTING [View Printable]
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user
Group: Member Posts: 29 Joined: Jul 07, 2005
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Some of the bands in my filter do not have a straight line compared to other bands even if they all are the same proteins. What can be wrong? One end of the band seemed to move faster than the the other end.
Thanks for any ideas!
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| Posted Sep 11, 2005, 13:18 PM |
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king of WB
Group: Member Posts: 6 Joined: Oct 28, 2005
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few things to consider: the same volume for all the samples cleaning your wells before loading and when loading try to load into the middle of the wells
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| Posted Oct 31, 2005, 14:43 PM |
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vanishing
Group: Member Posts: 130 Joined: Apr 25, 2005
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check the platinum wire in the gel running apparatus (consider replacing the platinum wire) do you have a homogenous electric field?
fill the empty slots with 1x sample buffer
does it slope only on one side (from one side to the other) or do you get a "smile"?
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| Posted Nov 01, 2005, 1:54 AM |
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Add colour 2 ur life
Group: Member Posts: 35 Joined: Sep 13, 2006
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hi there are so many factors 1.leaving the wells blank, try to fill the empty wells with the loading buffer try to avoid the LAST wells and load sample buffer same volume in that wells 2. try to run the gel in recommended volts dont choose higher voltage 3.use fresh gels dont use stored gels for PAGE 4.blott long time for low voltage
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| Posted Oct 07, 2006, 9:21 AM |
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yerasi.v. reddy
Group: Member Posts: 1 Joined: Oct 25, 2006
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| user said: | Some of the bands in my filter do not have a straight line compared to other bands even if they all are the same proteins. What can be wrong? One end of the band seemed to move faster than the the other end.
Thanks for any ideas! |
hi 1) Firstly check the salt concentration of u sample, donot exceed permissable salt concentration, 2) Secondly maintain uniform pH in all the samples(pHbasic) regards, venky
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| Posted Oct 26, 2006, 7:57 AM |
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mchinmoyee
Group: Member Posts: 104 Joined: Oct 08, 2008
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Dear User, I hope you might be getting proper bands by now , if not till now, then kindly scan the image and attach here, it will be easier to detect the accurate problem what you might be having. Wishes
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| Posted Oct 22, 2008, 3:53 AM |
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