Optimal working dilution should be determined by the investigator.
Protokoll
Positive Control: pSer / pThr Molecular Weight MarkerFormulation: The pSer/pThr molecular weight marker contains rabbit muscle phosphoproteins isolatedby Fe3+/IDA - affinity chromatography. Proteins are lyophilized fromPBS/NaF/PEG/Sucrose/ Bromophenolblue and Na - azide. After reconstitution the solutioncontains 0. 09% Na-azide. Stability: Reconstitute by addition of 200 µl H2O. After complete solubilization add 200 µl 2xSDS-PAGE sample buffer, mix and incubate at 90°C for 5 min. Application: The pSer/pThr molecular weight marker is recommended for immunoblot applications. Use20µl molecular weight marker per lane. Note: Use BSA based blot incubation buffers. Milk,Casein and Blotto might interfere with antibody - antigen interaction.
Beschränkungen
Nur für Forschungszwecke einsetzbar
Lagerung
-20 °C
Informationen zur Lagerung
Aliquote and store frozen. Avoid repeated freeze/thaw cycles. Shelf life: one year from despatch.
Phosphorylation and dephosphorylation of cellular proteins are central steps in transducing extracellular signals to the nucleus. Phosphorylated epitopes may serve as docking sites for the assembley of protein complexes or may alter the 3-dimensional protein structure thus modulating enzymatic activity or the ability to undergo protein-protein interactions. Modification of proteins on tyrosine residues is mediated by protein tyrosin kinases. Tyrosine phosphorylation may alter the biological activity or mediate the assembly of protein complexes via the interaction of phosphotyrosine residues with SH2 or PTB domains. Antibodies direct against phosphorylated epitopes recognize the phosphorylated amino acid in the context of the surrounding amino acid sequence. Recognition is therefore dependent on 2 criteria: 1) phosphorylation and 2) the surrounding amino acid motif. If one of the two criteria is not fulfilled, the antibody will not detect the phosphorylation site. Since the amino acid sequence varies between different phosphorylation sites, certain proteins - though phosphorylated - may not be detected by the antibody. Phosphorylation patterns in a given cell extract may differ when probed with different antibodies due to sequence specificity.