3. Predict the patterns (number of bands and apparent mol. wt) of the following

Question

3. Predict the patterns (number of bands and apparent mol. wt) of the following proteins on SDS gels: a) a monomeric protein with a molecular weight of 65,000 Da (65kDa) b) a trimeric protein containing three chains, each with a molecular weight of 60,000 Da c) immunoglobulin G in a non-reducing gel (no -mercaptoethanol added in the sample solution-the light chains have a molecular weight of 25,000 Da and the heavy chains 50,000 Da immunoglobulin G in a reducing gel (-mercaptoethanol added in the sample solution) d)

Explanation / Answer

3) a) A single band with a molecular weight of 65kDa will be observed when a monomeric protein with a molecular weight of 65kDA was ran on SDS PAGE.

b) A single band with a molecular weight of 60kDA will be observed because in SDS PAGE the interactions between three chains will be lost and they become monomers. So, a single thick band with a molecular weight of 60kDA will be seen.

c) A single band with a molecular weight of 150kDa will be observed on SDS PAGE because in non-reducing gel immunoglobulin will be intact and it has two light and two heavy chains. So, total molecular weight is 150kDa (25+25+50+50).

d) Two bands with molecular weight 50 and 25kDa will be seen on SDS PAGE because in reducing gel, disulfide linkages between heavy and light chains will be reduced and immunoglobulin will be separated as two heavy and two light chains. so, two bands ( heavy chain-50kDA and light chain-25kDa) will be seen on gel.

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