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Can the sequece-specific modifed peptide antibody used for affinity-enrichment?
Yes!
Only if this modified peptide antibody is able to immunoprecipitation (IP) the modified proteins.
No!
Not all the peptide antibodies are able to perform IP studies, even if it could be utilized for Western blotting (WB).
By using the pan-specific modified amino acid antibodies to affinity-enrich all the modified proteins, you may selectively detect the target modified proteins by WB using modified sequence-specific antibodies.
For WB, you could only resolve maximum 100 ug/lane of crude proteins in SDS-PAGE, your target modified proteins in 100 ug of crude lysates may not be detectable and may also interfere with other abundant proteins, but you could enrich your target modified proteins only from 1-10 mg of crude proteins, the enriched modified proteins could readily be resolved in SDS-PAGE and become detectable level for WB. |