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Nanog double band in WB


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#1 Stefano Giulitti

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Posted 24 April 2014 - 01:40 PM

Hi, i purchased an anti- human Nanog antibody from Reprocell (https://www.reprocel...man-nanog-poly/)

Nanog is expressed in pluripotent stem cells.

The antibody works ok by immunofluorescence. now I've tested it for western blot using an embryonic pluripotent stem cell line (HES2).

The datasheet shows an unique band at 37 kDa.

 

In my western (attached) 2 bands are visible. One at approx 42 kDa and one at higher values (60kDa).

Assuming a differential behavior during the PAGE depending on gel and buffer used, the 42 kDa can be the one indicated as 37 kDa.

 

Nanog dimerize, but SDS-PAGE should dissolve dimers (theoretically) and 42 kDa should give an 80 kDa band.

So what about the 60 kDa band?

 

thanks in advance

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#2 bob1

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Posted 28 April 2014 - 01:51 PM

Proteins are well known for not necessarily running at their predicted size - have a look at the antibody datasheet and see what info they provide.  You should also look at the literature and see what size their westerns are giving (if you can find the info).






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