Phosphoserine

Rabbit

IgG Polyclonal

Unconjugated. Also available conjugated with HRP, HRP.

Immunoaffinity purified

Unmodified

Phosphoserine conjugated with KLH.

Recognizes phosphoserine, peptidyl-phosphoserine and serine-phosphorylated proteins. Does not cross-react with ATP, phosphotyrosine, peptidyl phosphothreonine and serine. Slight cross-reactivity with free phosphothreonine. Readily reacts to known phosphoproteins such as phosvitin and alpha casein. Species cross-reactivity: All species.

• IHC (1:50)
• Western blot
• Immunoprecipitation
• ELISA (1:250 - 1:1000)

Suitable for use in Western Blot, Immunoprecipitation, Immunohistochemistry and ELISA. Western Blot (tissue lysates): 50-100 ug/lane total protein, antibody dilution can be from 1:250-1:1000, overloading will show solid exposed lanes. Dilute the primary antibody until multiple phosphorylated bands will appear. If using purified proteins dilutions from 1:50-1:1000 will need to be tried. At 1:300 dilution the antibody normally detects 10 ng of protein with 2 hour incubations and enhanced HRP detection. Blocking of the blot should be with non-phosphoserine containing compounds such as 1-5% BSA in TBS that has been filtered through a 0.45um. Milk or casein is not recommended because of the presence of phospho-serine in these solutions. Buffers should be Tris based so that phosphate ion interference is minimized. Immunoprecipitation (tissue extracts): 10-20 ug/mg protein. Note that this will immunoprecipitate all phosphoserine proteins in the extracts. If using purified protein exacts containing only the protein of interest, antibody amounts should be decreased to 5 ug/500 ul of extracts. ELISA (kinase assay): 1:250-1:500. Immunohistochemistry: 1:50. Detects all phosphoserines, and any phosphorylated serine protein or peptide can be used to block antibody staining. Typically, a10M excess of peptide is used in Tris based buffers. Positive control: NIH 3T3 cells (+/- TPA), K562 cells and EGF-stimulated A431 cells.

PBS, 50% Glycerol

For long-term storage and to avoid freeze-thaw cycles, aliquot and store at -20°C. Aliquots are stable for up to 1 year at -20°C.

For research use only. Intended for use by laboratory professionals.

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