Hello everyone,
I just finished a western today where I loaded cell lysates (each lane had the same cell line but with different plasmids in them) and I blotted using the same primary and secondary for all lanes. After development, I saw non-specic bands that were present in some lanes but not in others. Any idea to what is happening?!
Submit your paper to J Biol Methods today!
Non-specific bands different for same cell line, using same primary and secondar
Started by Elia El Habre, Sep 12 2012 07:52 AM
western non-specific molecular sds-page
1 reply to this topic
#2
bob1
-
- Global Moderators
-
- 5,108 posts
Thelymitra pulchella
327
Excellent
Excellent
Posted 12 September 2012 - 01:16 PM
Details please - how much protein or cells did you load per lane? What was the antibody against? Were the samples all collected at the same time? Were the samples all prepared in the same manner? Did you use (fresh) protease inhibitors?
Also tagged with one or more of these keywords: western, non-specific, molecular, sds-page
Protocols and Techniques Forums →
SDS-PAGE and Western Blotting →
Need help translating my western blot protocol into every languageStarted by doxorubicin, 03 Oct 2013  sds-page, western blot, language and 2 more...
|
|
|
||
Protocols and Techniques Forums →
SDS-PAGE and Western Blotting →
Problem with SDS-PAGE gelStarted by bio_film, 11 Aug 2013 sds-page, Gel, polyacrylamide and 1 more...
|
|
|
||
Protocols and Techniques Forums →
SDS-PAGE and Western Blotting →
How to dissolve TCA/acetone precipitated proteins?Started by Bacillus_Guy, 30 Jul 2013 precipitation, tca, acteone and 2 more...
|
|
|
||
Protocols and Techniques Forums →
SDS-PAGE and Western Blotting →
SDS-Page trouble-shootingStarted by Bacillus_Guy, 29 Jul 2013 sds-page, coomassie
|
|
|
||
Protocols and Techniques Forums →
PCR, RT-PCR and Real-Time PCR →
How to avoid nonspecific amplification (band) in PCRStarted by Inbox, 26 Jul 2013 PCR, non-specific
|
|
|
