Phosphothreonine Antibody (6K2H0)

Western Blot: Phosphothreonine Antibody (6K2H0) [NBP3-33576] - Western blot analysis of lysates from NIH 3T3 or NIH 3T3 treat with Calyculin A, using Phosphothreonine Rabbit mAb at 1:1000 dilution. NIH/3T3 cells were treated by Calyculin A (100 nM) at 37C for 30 minutes after serum-starvation overnight.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25ug per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Enhanced Kit.Exposure time: 60s.

Western Blot: Phosphothreonine Antibody (6K2H0) [NBP3-33576] - Western blot analysis of lysates from A-431, using Phosphothreonine Rabbit mAb at 1:1000 dilution. A 431 treated by Calyculin A (100 nM) at 37C for 30 minutes after serum-starvation overnight.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25ug per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Enhanced Kit.Exposure time: 60s.

• Reactivity: Hu, Mu
• Applications: WB, ELISA
• Clone: 6K2H0
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

Phosphothreonine Antibody (6K2H0) Summary

• Description: Novus Biologicals Rabbit Phosphothreonine Antibody (6K2H0) (NBP3-33576) is a monoclonal antibody validated for use in WB and ELISA. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: A synthetic phosphorylated peptide around of human Phosphothreonine.
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity: Affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA Recommended starting concentration is 1 ug/mL
  • Western Blot 1:500 - 1:1000

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: PBS (pH 7.3), 50% glycerol, 0.05% BSA
• Preservative: 0.05% Proclin 300
• Purity: Affinity purified

Alternate Names for Phosphothreonine Antibody (6K2H0)

• C4H10NO6P;O-phospho-L-threonine;Phospho-threonine;pThr;Threonine

Background

Phosphorylation of threonine residues is associated with many growth factors and oncogene protein kinases, and is important for cell signaling in activation, proliferation and differentiation. Protein phosphorylation and dephosphorylation are basic mechanisms for the modification of protein function in eukaryotic cells. Phosphorylation is a rare post-translational event in normal tissue, however, the abundance of phosphorylated cellular proteins increases several fold following various activation processes which are mediated through phosphotyrosine, phosphoserine or phosphothreonine (p-tyr/p-ser/p-thr). Many signal transduction pathways, such as the EGF, PDGF and insulin receptor systems, contain tyr/ser/thr kinase which phosphorylate specific tyr/ser/thr residues upon binding of ligands to their receptors. T cell antigen receptor complex or the receptors for some hemopoietic growth factors may stimulate these phosphorylation associated kinases, and cells transformed by viral oncogenes contain elevated levels of phosphorylated tyr/ser/thr. 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 their properties. 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. The most common procedure is to label intact cells or small tissue fragments with 32P and subsequently to isolate 32P labeled proteins by conventional biochemical methods. In order to identify the specific amino acids that undergo phosphorylation, additional long and tedious procedures for phosphoamino acid analysis are required. Immunoblotting of cellular proteins with antibodies directed against phosphoamino acids is advantageous as it does not involve 32P labeling, and can therefore be employed to monitor alterations in phosphorylation of specific proteins as they occur in intact organs or the whole animal. Indeed, mono and polyclonal antibodies directed against phosphorylated residues have been generated and found useful as analytical and preparative tools because they enable the rapid identification, quantification and immunoaffinity isolation of phosphorylated cellular proteins.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.