Why is the actual band size different from the predicted?
Western blotting is a technique that separates proteins based on size - in general, the smaller the protein the faster it migrates through the gel. However, migration is also affected by other factors and so the actual band size observed may differ from that predicted. Common factors include...
post-translational modification - e.g. phosphorylation, glycosylation etc which increases the size of the protein
post-translation cleavage - e.g. many proteins are synthesized as pro-proteins, and then cleaved to give the active form, e.g. pro-caspases
splice variants - alternative splicing may create different sized proteins from the same gene
relative charge - the composition of amino acids (charged vs non-charged)
multimers - e.g. dimerisation of a protein. This is usually prevented in reducing conditions, although strong interactions can result in the appearance of higher bands作者: queen 时间: 2013-5-13 15:55
Why is the actual band size different from the predicted?
Western blotting is a technique that separates proteins based on size - in general, the smaller the protein the faster it migrates through the gel. However, migration is also affected by other factors and so the actual band size observed may differ from that predicted. Common factors include...
post-translational modification - e.g. phosphorylation, glycosylation etc which increases the size of the protein
post-translation cleavage - e.g. many proteins are synthesized as pro-proteins, and then cleaved to give the active form, e.g. pro-caspases
splice variants - alternative splicing may create different sized proteins from the same gene
relative charge - the composition of amino acids (charged vs non-charged)
multimers - e.g. dimerisation of a protein. This is usually prevented in reducing conditions, although strong interactions can result in the appearance of higher bands
Notch1 (D1E11) XP® Rabbit mAb detects endogenous levels of total Notch1 protein. It recognizes both the full-length (~300 kDa) and the transmembrane/intracellular region NTM (~120 kDa).
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro2438 of human Notch1.
