I recently start to study how mgluR translocate from membrane to cytosol. I used mouse mgluR1 antibody from BD bioscience and found multiple bands (>250, 150 and 110). I heated up the sample at 60C and 5-6 min. The multiple bands observation does not agree with the info provided by BD , which is 133KDa.. I then prepared another batch of sample (heated up 95C for 5 min). The membrane fractions showed bands at 110KD band and cytosol fractions showed bands are 130KD . I am really confused here. Can anyone tell me what is going on? Which band is correct? Both are 7 % gels SDS-PAGE. First round used beta-mercaptoethanol, the second time I used DTT.
Should I use 60C or 95C to get the correct band for mGLUR1? I found some papers support the multiple bands observation, they didn't heat up the lysate at all, they said this is to avoid aggregate. Can anyone shed some light on this matter?
Hi ! Finally you solve the problem? I´m having the same issue. WB of hippocampus showed bands at 200 KD. Some papers describe a molecular weight about 125 KD for monomer and 250 KD for the dimer. I hope you can help me. Cheers!
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