Could you help me find a suitable loading control for microsomal proteins? We would like to do western blots on cyp1a1 and ugt. Beta-actin is commonly used in our lab, but I'm not sure if I can use it for microsomal fractions.
Use anti-KDEL (KDEL is the ER retention amino acid sequence)
Multiple ER resident proteins will be lit up with anti-KDEL (see below). Alternatively, you can blot for BiP (GRP78), which is a classical ER resident heat shock protein.
Below is information from Pierce:
The sequence Lys-Asp-Glu-Leu (KDEL) or a closely related sequence, is present at the carboxy-terminus of soluble endoplasmic reticulum (ER) resident proteins and some membrane proteins. 78 and 94 kDa glucose regulated proteins, GRP78 and GRP94 respectively, and protein disulfide isomerase (PDI) all share the C-terminal KDEL sequence. The presence of carboxy-terminal KDEL appears to be necessary for ER retention and appears to be sufficient to reduce the secretion of proteins from the ER. This retention is reported to be mediated by a KDEL receptor
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