Human MICA ELISA Kit (ab100592)

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Overview

  • Product name

    Human MICA ELISA Kit
    See all MICA kits

  • Detection method

    Colorimetric

  • Sample type

    Cell culture supernatant, Serum, Plasma

  • Assay type

    Sandwich (quantitative)

  • Sensitivity

    < 20 pg/ml

  • Range

    13.72 pg/ml - 10000 pg/ml

  • Recovery

    100 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 116.9 109% - 124%
    Serum 112.3 95% - 129%
    Plasma 93.83 75% - 107%

  • Assay duration

    Multiple steps standard assay

  • Species reactivity

    Reacts with: Human

  • Product overview

    Abcam’s MICA Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme linked immunosorbent assay for the quantitative measurement of Human MICA in serum, plasma, and cell culture supernatants.

    This assay employs an antibody specific for Human MICA coated on a 96-well plate. Standards and samples are pipetted into the wells and MICA present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human MICA antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin 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 MICA bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Notes

    Optimization may be required with urine samples.

  • 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
    300X HRP-Streptavidin Concentrate 1 x 200µl
    5X Assay Diluent 1 x 15ml
    Biotinylated anti-Human MICA (lyophilized) 2 vials
    MICA Microplate (12 x 8 wells) 1 unit
    Recombinant Human MICA Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml

  • Research areas

  • Relevance

    The MHC class I chain-related (MIC) proteins are related to the Major histocompatibility complex (MHC) class I proteins which are ubiquitously expressed and mediate the recognition of intracellular antigens by cytotoxic T cells. The MHC class I chain-related (MIC) proteins are recognized by NKG2D, a receptor on NK and T cells, and promote anti-tumor activity. MICA, a member of the MIC family, is widely expressed on many tumors, and it is the MICA/NKG2D interaction that is thought to stimulate the anti-tumor reactivity by T lymphocytes. MICA is present in virtually every tissue except the nervous system, suggesting that MIC protein expression may only be one component of the anti-tumor activity of the immune system.

  • Cellular localization

    Plasma membrane

  • Alternative names

    • FLJ60820
    • HLA class I antigen
    • MGC111087
    • MHC class I chain related gene A protein
    • MHC class I chain related protein A
    • MHC class I polypeptide related sequence A
    • MHC class I related protein
    • MIC A
    • PERB11.1
    • Stress inducible class I homolog
    see all

  • Database links

    Applications

    Our Abpromise guarantee covers the use of ab100592 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

    • ELISA: MICA Human ELISA Kit (ab100592)
      ELISA: MICA Human ELISA Kit (ab100592)

      Standard curve with background signal subtracted (duplicates; +/- SD).

    • ELISA: MICA Human ELISA Kit (ab100592)
      ELISA: MICA Human ELISA Kit (ab100592)

      MICA measured in undiluted cell culture supernatants, U937 and THP-1 signals were below level of detection (13.7 pg x mL-1) (duplicates +/- SD).

    • Typical Standard Curve
      Typical Standard Curve

      Representative Standard Curve using ab100592.

    Protocols

    References

    This product has been referenced in:

    • Ren J  et al. Estrogen upregulates MICA/B expression in human non-small cell lung cancer through the regulation of ADAM17. Cell Mol Immunol N/A:N/A (2014). Read more (PubMed: 25363527) »
    See 1 Publication for this product

    Publishing research using ab100592? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Question

    1) Extracellular domain of MICA not detected. Is the standard the recombinant full length MICA protein or a fragment?
    2) Standard values are much lower than the ones obtained with a kit from a competitor. The values are ok when the standard from the second kit is used. When the standard from Abcam's kit is used with the second kit the values are low. Conclusion: problem of weakness with the Abcam's standard.

    Read More
    Answer

    Thank you for contacting us.

    Please find below the answers from our collaborator to your questions:

    1) Extracellular domain of MICA not detected. Is the standard the recombinant full length MICA protein or a fragment?
    The antibody pair in the kit was raised against the extracellular domain of MICA (aa 24-307). The recombinant standard protein included in the kit is also just the extracellular portion.


    2) Standard values are much lower than the ones obtained with a kit from a competitor. The values are ok when the standard from the second kit is used. When the standard from Abcam's kit is used with the second kit the values are low. Conclusion: problem of weakness with the Abcam's standard.
    The weaker signal from testing our recombinant standard protein on another manufacturer’s kit could be the result of an issue with the protein but it could also be due to different antibodies being used which recognize different binding sites, difference in sensitivities, different buffers and diluents used, etc.
    Therefore with all of these variables to consider when mixing components between two different kits, it would be difficult to pin point the exact cause as there could be many contributing factors.

    At what temperature was the standard stored at upon receipt? For how long?
    Once reconstituted to 100ng/ml was this stored for later use? If so, at what temperature and for how long?
    Any freeze-thaw cycles? If so, how many?
    How was the standard prepared? Vortexing? By pipette?

    Moving forward I would remind of the storage guidelines in the manual and the fact that an extra vial of lyophilized standard (2 vials total) is included in each kit to minimize storage issues. Also, attached are some additional tips on ensuring a strong standard signal strength that the customer should keep in mind and find useful.


    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. Our collaborator
    would be happy to take a look at the raw numerical absorbance data though if you would like.

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    Question

     Dear tech,   A customer would like to purchase a ELISA kit for soluble MICA. Do ab100592 and ab59569 match the need? Thank you for your help!   Best regards,  

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    Answer

      Thank you for your patient. Both products are suitable to use with soluble MICA.  However, please notice there are slightly differences between the kits. Ab100592 has been tested in human fluids, as Urine, Serum, and Plasma and also in Cell culture supernatant. Ab59569 requires cell culture supernatants and cell culture extracts as sample types. The sensitivity and the range are also different depending on the kit. I recommend you to have a look at their datasheets and decide which one fulfills your requirements. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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