Phosphoserine

Antibodies / Antibodies-Amino Acids

-20°C

Supplied as a liquid in 0.01M PBS, pH 7.4, 15mM sodium azide.

Phosphoserine conjugated to KLH.

Protein phosphorylation and dephosphorylation are basic signaling mechanisms that modify protein function in eukaryotic cells.4 Phosphorylation is a rare post- translational event in normal tissues, however, the abundance of phosphorylated cellular proteins increases several fold following various activation processes. The main amino acids that are phosphorylated are tyrosine, serine, or threonine (pTyr/pSer/pThr), each having specific kinases that phosphorylate them and specific phosphatases that dephosphorylate them. Different growth factor receptors, such as the EGF-R, PDGF-R, and insulin receptor, contain Tyr/Ser/Thr residues that are autophosphorylated upon binding to their ligands. Ligand binding of other receptors, like the T cell antigen receptor complex or receptors for some hematopoietic growth factors, stimulate downstream associated kinases.6 An understanding of transformation by oncogenes and mitogenic processes of growth factors depends on the identification of their substrate and a subsequent determination of how phosphorylation affects the properties of these proteins. Studies on the role of phosphorylated proteins have been hampered by their low abundance and the problem of distinguishing the various types of phosphorylated proteins. Immunoblotting of cellular proteins with antibodies directed against phosphoamino acids is advantageous over 32P labeling, and can therefore be employed to monitor alterations in phosphorylation of specific proteins as they occur in intact organs or even in whole animals. Indeed, mono- and polyclonal antibodies directed against phosphorylated residues were generated and found useful as analytical and preparative tools 2, 7 by enabling the identification, quantification and immunoaffinity isolation of phosphorylated cellular proteins.

Applications:
Suitable for use in Indirect ELISA, Western Blot and Dot Blot. Other applications not tested.

Recommended Dilution:
ELISA (indirect): 0.3-0.6ug/ml using phosphoserine conjugated to BSA
Western Blot (indirect): 2.5-5ug/ml using total rat brain extract
Note: Do not use milk as a blocking agent or in diluents, as milk casein is phosphorylated at several serine residues. BSA is recommended instead.
Optimal dilutions to be determined by researcher.

Hybridoma:
Mouse myeloma cells with splenocytes from Balb/c mice.

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.