Overview

  • Product name

    Human PRAME ELISA Kit
    See all PRAME kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Cell extract 5 4.9%
    Inter-assay
    Sample n Mean SD CV%
    Cell extract 3 3%
  • Sample type

    Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    83 pg/ml
  • Range

    0.313 ng/ml - 20 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 105 104% - 106%
    Tissue Extracts 95 94% - 96%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    PRAME in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of PRAME protein in human cell and tissue extracts.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB Development Solution is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    PRAME (preferentially expressed antigen in melanoma) functions as a transcriptional repressor, inhibiting the signaling of retinoic acid through the retinoic acid receptors RARA, RARB and RARG. PRAME prevents retinoic acid-induced cell proliferation arrest, differentiation and apoptosis. PRAME is a cancer/testis antigen not normally expressed in any tissues except testis. PRAME is upregulated in melanoma cells and it is recognized by cytolytic T-cells. PRAME expression is upregulated in other tumors, such as synovial sarcoma, NSCLC and breast cancer, where it contributes to tumorigenesis and metastasis. PRAME and other cancer/testis antigens are currently being pursued as novel immunotherapy targets and diagnostic biomarkers.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human PRAME Capture Antibody 1 x 600µl
    10X Human PRAME Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml
    Denaturant 1 x 500µl
    Human PRAME Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Functions as a transcriptional repressor, inhibiting the signaling of retinoic acid through the retinoic acid receptors RARA, RARB and RARG. Prevents retinoic acid-induced cell proliferation arrest, differentiation and apoptosis.
  • Tissue specificity

    Expressed in testis. Detected in samples of kidney, brain and skin.
  • Sequence similarities

    Belongs to the PRAME family.
    Contains 4 LRR (leucine-rich) repeats.
  • Cellular localization

    Nucleus. Cell membrane.
  • Information by UniProt
  • Alternative names

    • 4930534P07Rik
    • Cancer/testis antigen 130
    • CT130
    • MAPE
    • Melanoma antigen preferentially expressed in tumors
    • OIP 4
    • OIP-4
    • OIP4
    • OPA interacting protein 4
    • Opa interacting protein OIP4
    • OPA-interacting protein 4
    • PRAME
    • PRAME_HUMAN
    • Preferentially expressed antigen in melanoma
    • Preferentially expressed antigen of melanoma
    • RP23-250F8.3
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab234561 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

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • The PRAME standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of PRAME were measured in duplicate and interpolated from the PRAME standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PRAME concentration was determined to be 14.70 ng/mL in K562 cell extract and 2.359 ng/mL in testis tissue extract.

Protocols

References

ab234561 has not yet been referenced specifically in any publications.

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