Immunoblotting (Western Blot): 1 μg/mL for HRPO/ECL detection. Recommended blocking buffer: BSA/Tween 20 basedblocking buffer. DO NOT USE MILK OR CASEIN FOR BLOCKING! ELISA: 0.05 μg/mL. Immunoprecipitation: 1-10 μg per 10^6 pervanadate-treated A431 cells. Positive Control: Phosphoserine/Phosphothreonine. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protocol
Positive Control: pSer / pThr Molecular Weight MarkerFormulation: The pSer/pThr molecular weight marker contains rabbit musclephosphoproteins isolated by Fe3+/IDA - affinity chromatography. Proteins are lyophilizedfrom PBS/NaF/PEG/Sucrose/ Bromophenolblue and Na - azide. After reconstitution thesolution contains 0. 09% Na-azide. Stability: Reconstitute by addition of 200 µl H2O. After complete solubilization add 200 µl2x SDS-PAGE sample buffer, mix and incubate at 90°C for 5 min. Application: The pSer/pThr molecular weight marker is recommended for immunoblotapplications. Use 20µl molecular weight marker per lane. Note: Use BSA based blotincubation buffers. Milk, Casein and Blotto might interfere with antibody - antigeninteraction. Storage: Aliquote and store frozen. Avoid repeated freeze/thaw cycles. Shelf life: one year from despatch.
Restrictions
For Research Use only
Reconstitution
Restore with 1 mL H2O (15 min, RT).
Buffer
1 mL2 x PBS / 0.09 % Sodium Azide / PEG and Sucrose.
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
-20 °C/-80 °C
Storage Comment
Store lyophilized (preferably in a desiccator) at -20 °C and reconstituted (aliquote andfreeze in liquid nitrogen) at -80 °C. Avoid repeated freezing and thawing. Thaw aliquots at 37 °C. Thawed aliquots may be stored at 4 °C up to 1 week. Shelf life: one year from despatch.
Phosphorylation and dephosphorylation of cellular proteins are central steps in transducing extracellular signals to the cell nucleus. Phosphorylated epitopes may serve as docking sites for the assembly 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 serine residues is mediated by serine/threonine kinases.