Overview

  • Product name

    Met (pY1234/pY1235) + total Met ELISA Kit
    See all Met (c-Met) kits
  • Detection method

    Colorimetric
  • Sample type

    Cell Lysate, Tissue Lysate
  • Assay type

    Semi-quantitative
  • Assay time

    5h 00m
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Product overview

    ab126451 is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human and mouse cell lysates. By determining phosphorylated Met protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.

    This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of phospho-Met (Tyr1234/1235) and total Met in human, and mouse cell lysates (help normalize the results of phospho-Met from different cell lysate being compared). A pan Met antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and Met present in a sample is bound to the wells by the immobilized antibody. The wells are washed and anti-phospho-Met (Tyr1234/1235) or anti-pan-Met is used to detect phosphorylated or total Met. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Met (Tyr1234/1235) or total Met bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    2X Cell Lysis Buffer 1 x 5ml
    5X Assay Diluent 1 x 15ml
    Detection Antibody Met 1 vial
    Detection Antibody Met (Y1234/1235) 1 vial
    500X HRP-conjugated anti-rabbit IgG 1 x 25µl
    Met Microplate (12 strips x 8 wells) coated with anti-pan Met antibody 1 unit
    Positive Control: lyophilized powder from H1993 cell lysate 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas

  • Function

    Receptor for hepatocyte growth factor and scatter factor. Has a tyrosine-protein kinase activity. Functions in cell proliferation, scattering, morphogenesis and survival.
  • Involvement in disease

    Note=Activation of MET after rearrangement with the TPR gene produces an oncogenic protein.
    Note=Defects in MET may be associated with gastric cancer.
    Defects in MET are a cause of hepatocellular carcinoma (HCC) [MIM:114550].
    Defects in MET are a cause of renal cell carcinoma papillary (RCCP) [MIM:605074]. It is a subtype of renal cell carcinoma tending to show a tubulo-papillary architecture formed by numerous, irregular, finger-like projections of connective tissue. Renal cell carcinoma is a heterogeneous group of sporadic or hereditary carcinoma derived from cells of the proximal renal tubular epithelium. It is subclassified into common renal cell carcinoma (clear cell, non-papillary carcinoma), papillary renal cell carcinoma, chromophobe renal cell carcinoma, collecting duct carcinoma with medullary carcinoma of the kidney, and unclassified renal cell carcinoma.
    Note=A common allele in the promoter region of the MET shows genetic association with susceptibility to autism in some families. Functional assays indicate a decrease in MET promoter activity and altered binding of specific transcription factor complexes.
    Note=MET activating mutations may be involved in the development of a highly malignant, metastatic syndrome known as cancer of unknown primary origin (CUP) or primary occult malignancy. Systemic neoplastic spread is generally a late event in cancer progression. However, in some instances, distant dissemination arises at a very early stage, so that metastases reach clinical relevance before primary lesions. Sometimes, the primary lesions cannot be identified in spite of the progresses in the diagnosis of malignancies.
  • Sequence similarities

    Belongs to the protein kinase superfamily. Tyr protein kinase family.
    Contains 3 IPT/TIG domains.
    Contains 1 protein kinase domain.
    Contains 1 Sema domain.
  • Domain

    The kinase domain is involved in SPSB1 binding.
  • Post-translational
    modifications

    Dephosphorylated by PTPRJ at Tyr-1349 and Tyr-1365.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Alternative names

    • AUTS9
    • c met
    • D249
    • Hepatocyte growth factor receptor
    • HGF
    • HGF receptor
    • HGF/SF receptor
    • HGFR
    • MET
    • Met proto oncogene tyrosine kinase
    • MET proto oncogene, receptor tyrosine kinase
    • Met proto-oncogene
    • Met proto-oncogene (hepatocyte growth factor receptor)
    • Met protooncogene
    • MET_HUMAN
    • Oncogene MET
    • Par4
    • Proto-oncogene c-Met
    • RCCP2
    • Scatter factor receptor
    • SF receptor
    • Tyrosine-protein kinase Met
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab126451 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • H1993 cells were cultured at 37°C for 4 days. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed by Western blot.
  • H1993 cells were cultured at 37°C for 4 days. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed using ab126451.
  • A431 cells were treated or untreated with 50 ng/ml recombinant human HGF for 5 min. Cell lysates were analyzed using ab126451.
  • H1993 cells were cultured at 37°C for 4 days. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed using ab126451.

Protocols

References

This product has been referenced in:

  • AbouAitah K  et al. Folic acid-conjugated mesoporous silica particles as nanocarriers of natural prodrugs for cancer targeting and antioxidant action. Oncotarget 9:26466-26490 (2018). Read more (PubMed: 29899871) »
See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer



It is unusual that the positive control would not dissolve. It is just a lyophilized H1993 cell lysate.

However please note the positive controls will not be used for any calculations with the samples as the kit provides semi-quantitative results. The positive control is essentially a fail-safe to let you know the kit is working if you do not receive good results from your samples. As you are getting good results with your own samples, the kit is working fine and you can go ahead with the assay.

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Answer

This kit is compatible with tissue samples. I am contacting our datasheets team to have tissue lysates added to the sample type section of the datasheet, since we will guarantee this kit to work with tissue. I have attached our general recommended tissue preparation protocol which may be of help to you. You may use the lysis buffer provided with the kit for homogenization.

We do recommend using our protease inhibitor cocktail ab65621 (link below):

https://www.abcam.com/protease-inhibitor-cocktail-ab65621.html

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Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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