Phosphoserine\n

Reversible protein phosphorylation plays a central role in numerous biochemical pathways and functions to alter protein conformation and activity.(6,7) Detection of protein phosphorylation is frequently accomplished by incorporation of exogenous 32P(1,5) followed by amino acid analysis.(1,5,6,7) In an effort to simplify this process, specific antibodies to Phosphotyrosine (PY) were initially developed(3,4), some with the ability to detect a single phosphorylated tyrosine residue.(3,4,8) While the production of anti-phosphotyrosine antibodies has been relatively straight forward, the structural similarity of phosphoserine (PS) and phosphothreonine (PT) has made production of specific antibodies to these amino acids more problematic.(5)\n\nApplications: \nSuitable for use in ELISA, Western Blot and Immunoprecipitation. Other applications not tested.\n\nRecommended Dilution:\nELISA: 0.1-1.0ug.ml.\nWestern Blot: 1-2ug/ml. Milk-derived blocking solutions reportedly contain phosphoproteins that may inhibit phosphoamino acid antibody binding and therefore should be avoided. A 3% BSA solution may be used.\nImmunoprecipitation: 3-5ug/IP reaction\nOptimal dilutions to be determined by the researcher.\n\nPositive Control:\nNIH 3T3 cells (+/-TPA), K562 cells, EGF-stimulated A431 cell lysates.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile 40-50% glycerol, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

P4076-50A

Size

50ug

Applications

ELISA, IP, WB

Hosts

Rabbit

Form

Supplied as a liquid in PBS, pH 7.4, 0.1% sodium azide.

P Type

Pab

Purity

Purified by affinity chromatography.

Isotype

IgG

References

1. Edelman, AM., et al., Annu. Rev. Biochem. 56:567(1987). 2. Fleming IN, et al. J. Biol. Chem. 272(52): 33105-33110 (1997). 3. Frackelton, AR., et al., Mol. Cell. Biol. 3:1343 (1983). 4. Glenny, JR., et al., J. Immunological Meth. 109:277 (1988). 5. Heffetz, D., et al., Methods in Enzymology 210:44 (1991). 6. Hunter, T. Methods in Enzymology 200, 3 (1991). 7. Hunter, T Cell 50, 823 (1987). 8. Sengupta, A., et al., Proc. Natl. Acad. Sci. USA 85:8062 (1988).

Additional Info

Recognizes proteins containing phosphorylated serine residues (pSer). Recognition is independent of neighboring amino acids and species of origin of the phosphorylated protein. No cross-reactivity to proteins phosphorylated on threonine or tyrosine residues.