Weird - shaped band on western blotting

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namakulaila
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Weird - shaped band on western blotting

Post by namakulaila » Thu Oct 17, 2013 6:17 am

Hi everyone,

This is my first post in this forum.
My name is Laila and currently working with the hepatoma cells AH109A to see the cell cycle regulation protein.

Recently I got a weird-shaped band on my western blot. It is like an upside-down of smiling face in each lane. I use the same loading buffer, electrophoresis buffer, lysis buffer, and also the same 1st and 2nd antibody for this experiment. I also try to reduce the amount of protein in each lane to 25μg, and work with 15mA during electrophoresis but still no change.

Is there anybody had the same problem previously?
I attached the tubulin band for reference


Regards,
Laila
Attachments
tubulin.tif
weird- shaped tubulin band
(1.5 MiB) Not downloaded yet

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JackBean
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Post by JackBean » Thu Oct 17, 2013 2:17 pm

are you sure you didn't put your gel upside down?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

namakulaila
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Post by namakulaila » Fri Oct 18, 2013 4:09 am

Yes, I am sure I put it on the right position, since I always mark upper side of the gel

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