Overview

  • Product name

    Anti-Phosphothreonine antibody [RM102]
    See all Phosphothreonine primary antibodies
  • Description

    Rabbit monoclonal [RM102] to Phosphothreonine
  • Host species

    Rabbit
  • Specificity

    ab238305 reacts threonine-phosphorylated proteins. No cross reactivity with nonphosphorylated threonine, phosphoserine, and phosphotyrosine. It shows slight cross-reactivity with a few phospho-serine-containing peptides.
  • Tested applications

    Suitable for: WB, IP, ICC/IFmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Synthetic peptide corresponding to Phosphothreonine conjugated to bovine serum albumin. Mixture of phosphothreonine-BSA conjugate and a phosphothreonine containing peptide.

  • Positive control

    • ICC/IF: Serum-starved A431 cells treated with Calyculin A/Okadaic Acid. WB: Serum-starved A431 cells treated with Calyculin A/Okadaic Acid. IP: A431 cells treated with Calyculin A/Okadaic Acid.

Properties

Applications

Our Abpromise guarantee covers the use of ab238305 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/500 - 1/2000.
IP 1/100 - 1/500.
ICC/IF 1/100 - 1/500.

Target

  • Relevance

    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.

Images

  • Serum-starved A431 (human epidermoid carcinoma cell line) cells treated with Calyculin A/Okadaic Acid stained for Phosphothreonine using ab238305 at a 1/500 dilution in ICC/IF, followed by a PE conjugated secondary antibody (red). Counterstained with DAPI (blue).

    Left panel: Non-treated cells.

    Right panel: Cells treated with Calyculin A/Okadaic Acid.

  • All lanes : Anti-Phosphothreonine antibody [RM102] (ab238305) at 1/2000 dilution

    Lane 1 : Serum starved A431 (human epidermoid carcinoma cell line) cell lysate
    Lane 2 : Serum starved A431 cells treated with Calyculin A/Okadaic Acid lysate
  • Immunoprecipitation of Calyculin A/Okadaic Acid treated A431 (Human epidermoid carcinoma cell line) cells by ab238305 at a 1/500 dilution, and then blotted with ab238305.

    Lane 1: Whole lysate control.
    Lane 2: IP with rabbit IgG control.
    Lane 3: IP with ab238305.

  • ab238305 recognizes Phosphorylated threonine in peptides with different sequences.

    It has minimal cross-reactivity with phosphorylated serine.

References

ab238305 has not yet been referenced specifically in any publications.

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