Overview

  • Product name

    Human PAPPA ELISA Kit
    See all PAPP A kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 3 4.7%
    Inter-assay
    Sample n Mean SD CV%
    Serum 8 5.7%
  • Sample type

    Serum, Tissue Extracts, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    8.89 pg/ml
  • Range

    156.25 pg/ml - 14000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 109 103% - 115%
    Tissue Extracts 105 100% - 112%
    Heparin Plasma 96 95% - 96%
    EDTA Plasma 95 85% - 113%
    Citrate Plasma 86 84% - 87%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat
  • Product overview

    PAPPA in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of PAPPA protein in human serum, plasma, 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.


    Human PAPPA is also called Pregnancy Associated Plasma Protein A or pappalysin 1 is a metalloproteinase which specifically cleaves IGFBP-4 and IGFBP-5, resulting in release of bound IGF. Cleavage of IGFBP-4 is dramatically enhanced by the presence of IGF, whereas cleavage of IGFBP-5 is slightly inhibited by the presence of IGF. High levels of PAPPA are found in placenta and pregnancy serum. In placenta, PAPPA is specifically expressed in X cells in septa and anchoring villi, and in syncytiotrophoblasts in the chorionic villi. Lower levels are found in a variety of other tissues including kidney, myometrium, endometrium, ovaries, breast, prostate, bone marrow, colon, fibroblasts and osteoblasts, Mouse and rat both share 91% sequence homology with the human PAPPA.


    Sensitivity


    Samples diluted in Sample Diluent NS - 8.89 pg/mL
    Samples diluted in 1X Cell Extraction Buffer - 67.21 pg/mL

  • 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 PAPPA Capture Antibody 1 x 600µl
    10X Human PAPPA Detector Antibody 1 x 600µl
    Human PAPPA Lyophilized Recombinant Protein 2 vials
    Antibody Diluent 4BI 1 x 6ml
    10X Wash Buffer PT (ab206977) 1 x 20ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    50X Cell Extraction Enhancer Solution 1 x 1ml
    TMB Development Solution 1 x 12ml
    Stop Solution 1 x 12ml
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Plate Seals 1 unit
  • Research areas

  • Function

    Metalloproteinase which specifically cleaves IGFBP-4 and IGFBP-5, resulting in release of bound IGF. Cleavage of IGFBP-4 is dramatically enhanced by the presence of IGF, whereas cleavage of IGFBP-5 is slightly inhibited by the presence of IGF.
  • Tissue specificity

    High levels in placenta and pregnancy serum. In placenta, expressed in X cells in septa and anchoring villi, and in syncytiotrophoblasts in the chorionic villi. Lower levels are found in a variety of other tissues including kidney, myometrium, endometrium, ovaries, breast, prostate, bone marrow, colon, fibroblasts and osteoblasts.
  • Sequence similarities

    Belongs to the peptidase M43B family.
    Contains 5 Sushi (CCP/SCR) domains.
  • Developmental stage

    Present in serum and placenta during pregnancy; levels increase throughout pregnancy.
  • Post-translational
    modifications

    There appear to be no free sulfhydryl groups.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • ASBABP2
    • Aspecific BCL2 ARE binding protein 2
    • Differentially placenta 1 expressed protein
    • DIPLA1
    • EC 3.4.24.79
    • IGF dependent IGFBP 4 protease
    • IGF-dependent IGFBP-4 protease
    • IGFBP 4ase
    • IGFBP-4ase
    • IGFBP4ase
    • Insulin like growth factor dependent IGF binding protein 4 protease
    • Insulin-like growth factor-dependent IGF-binding protein 4 protease
    • PAPA
    • PAPP A
    • PAPP-A
    • PAPP1_HUMAN
    • PAPPA
    • PAPPA1
    • Pappalysin 1 precursor
    • Pappalysin-1
    • Pregnancy Associated Plasma Protein A
    • Pregnancy associated plasma protein A pappalysin 1
    • Pregnancy-associated plasma protein A
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab235647 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.

     

  • Standard curve comparison between Human PAPPA SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of PAPPA were measured in duplicates, interpolated from the PAPPA standard curves and corrected for sample dilution. Undiluted samples are as follows: pregnant serum 1:500 and pregnant serum pre-eclampsia 1:500. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PAPPA concentration was determined to be 1669.24 ng/mL in pregnant serum, and 2393.50 ng/mL in pregnant serum pre-eclampsia.

  • The concentrations of PAPPA were measured in duplicate and interpolated from the PAPPA standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PAPPA concentration was determined to be 2862.86 pg/mL.

Protocols

References

ab235647 has not yet been referenced specifically in any publications.

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