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help with protein size

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1 reply to this topic

#1 cyndi



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Posted 21 September 2004 - 08:41 AM

Hi all,

Hope someone has a suggestion here. I purchased an antibody from a company whose technical support said that the specificity of the antibody was confirmed by running a Western. Their band showed up around 105-105 kDa. I ran a Western with the antibody (both rat tissue which they used and my test tissue) and I am getting bands for both sets of tissue at about 50-52 kDa. I have tried using protease inhibitors, both an recipe and a inhibitor cocktail, and still have similar results. The antibody is a polyclonal, but I'm not sure how likely it is that it is now recognizing a different protein since it has already been tested in rat with a single band at 105. Also, FYI the protein is a dimer, but literature suggests that each subunit should be about 100-110 kDa, not the dimer. Any suggestions or explanations as to why this protein would be half the size of the normal?

#2 mujan



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Posted 23 September 2004 - 11:27 AM

Could you be seeing the heavy chain of the IgG? Have you tried a longer exposure to see if something is present in the 105 range? Try with and w/o BME or boiling.

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