• Product name

    Anti-Phosphothreonine antibody (HRP)
    See all Phosphothreonine primary antibodies
  • Description

    Rabbit polyclonal to Phosphothreonine (HRP)
  • Host species

  • Conjugation

  • Specificity

    Reacts with free phosphothreonine but does not react with phosphoserine, threonine or phosphotyrosine.
  • Tested applications

    Suitable for: WB, IP, ELISAmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Chemical/ Small Molecule conjugated to KLH.

  • Positive control

    • Use mouse brain extract for immunoblotting. Use synthetic phosphopeptide (on threonine) for ELISA.



Our Abpromise guarantee covers the use of ab9338 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
  • Application notes
    ELISA(kinase assay): Use at 0.5 µg/mL
    Western blot: Use at 4µg/mL
    IP: Use at 10 µg/250 µg protein sample
    Will detect 100 ng of phosvitin in Western Blots and 0.5 ng of phosvitin with ELISA.
    Can be used for non-radioactive protein kinase assay (ELISA) using biotinylated peptide substrate and immunoblotting of abundant phosphoproteins.
    It is not recommended for immunoblotting of trace cellular phosphoproteins.
    Acetone precipitation of the protein extract followed by SDS denaturation is recommended for successful immunoprecipitation.
  • 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.


    • Immunoblotting of fetal mouse brain extract (125 ug - A and 25 ug - B)

      Immunoblotting of fetal mouse brain extract (125 ug - A and 25 ug - B)
    • Antibody Capture ELISA
      Label: immobilized antigen


    This product has been referenced in:

    • Virok DP  et al. Chlamydial infection induces pathobiotype-specific protein tyrosine phosphorylation in epithelial cells. Infect Immun 73:1939-46 (2005). WB . Read more (PubMed: 15784533) »
    See 1 Publication for this product

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A


    Thank you for your reply. It is very good to know that plenty of phosphatase inhibitors are present in your lysis buffer. I would recommend to incubate the antibody overnight at 4C if you do not do so already, and indeed the blocking step in BSA should help too. If you are still unhappy with your results with those changes please do not hesitate to contact me again and I can offer you a replacement vial or refund if the antibody was purchased in the last 90days,

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    I'm very sorry to hear you are experiencing problems with ab9338. You mention having tried a range of concentrations of primary antibody, could you clarify what incubation times you have tried? The blocking buffer milk is not recommended for phospho-specific antibodies, I would therefore recommend to use 5%BSA in TBST (30min- 1hr) prior to the antibody incubation in TBST only. I am wondering if your levels of phospho-threonine are high enough in your lysates. Could the phospho group be destroyed by phosphatases i.e does the complete inhibitor contain any phosphatase inhibitors such as sodium fluoride and sodium orthovanadate? Many thanks for providing this information which will help me understand your problem better, I apologise for the delay in resolving this matter and look forward to hearing from you,

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