Hi, I´m doing Western Blot and normalizing with GAPDH.
I usually perform it with mouse skeletal muscle samples. I am using GAPDH (D16H11) XP Rabbit mAb #5174 from CellSignalling with an antirabbit secondary Ab.
The problem is that I use to see 2 bands at 37 and 45 kDa when it is indicated to give only 1 band at 37 kDa. More often is the upper band more intense and homogeneous between samples but it sometimes happens with the lower band.
Does anyone know what could be happening? Thanks
Hello Gabriel,
They could be isoforms in order to meet various developmental, physiological, or pathological needs. You may want to refer to the links attached below for more information.
https://www.degruyter.com/document/doi/10.1515/biol-2021-0130/html?lang=en
https://www.uniprot.org/uniprotkb/P16858/entry#sequences
https://www.ncbi.nlm.nih.gov/gene?Db=gene&Cmd=DetailsSearch&Term=14433
https://www.genecards.org/cgi-bin/carddisp.pl?gene=GAPDH
Best.
Hi
as you'll see in the attached picture, there are 2 isoforms differing at 5' of the gene.
all the best
fred
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