Overview

  • Product name

    Human Vitamin D Binding Protein ELISA Kit
    See all Vitamin D Binding protein kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 6 3%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 4.3%
  • Sample type

    Cell culture supernatant, Saliva, Milk, Urine, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    2.8 pg/ml
  • Range

    19.5 pg/ml - 1250 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Saliva 99 86% - 107%
    Milk 102 93% - 108%
    Urine 102 99% - 108%
    Serum 95 91% - 99%
    Cell culture media 95 95% - 95%
    Heparin Plasma 92 88% - 96%
    EDTA Plasma 101 94% - 105%
    Citrate Plasma 98 94% - 102%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

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

    Vitamin D Binding Protein in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Vitamin D Binding Protein in serum, plasma, saliva, urine, milk, and cell culture supernatant.


    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 substrate 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.


    Vitamin D Binding Protein, also known as gc-globulin, is a 458 - amino acid, glycosylated protein encoded by the gene GC, which is in the albumin gene family. The primary function of this protein is to bind Vitamin D and its metabolites and to transport them to tissue. Vitamin D Binding protein is highly expressed and secreted into serum primarily from hepatic cells. There are three common alleles of the protein (Gc1f, Gc1s, and Gc2), and several minor polymorphisms. Different glycosylation sites are found on the isoforms of Vitamin D binding protein, a selectively deglycosylated form produced by B cell β-galactosidase and T cell sialidase is known as Macrophage Activating Factor. MAF has been found to block angiogenic effects of several proteins on vascular endothelial cells. 

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

Applications

Our Abpromise guarantee covers the use of ab223586 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 Vitamin D Binding Protein SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.

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

  • The concentrations of Vitamin D Binding Protein were measured in duplicates, interpolated from the Vitamin D Binding Protein standard curves and corrected for sample dilution. All undiluted samples start at 1:500,000 in Sample Diluent NS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Vitamin D Binding Protein concentration was determined to be 427 µg /mL in neat serum, 380 µg /mL in neat plasma (citrate), 552 µg /mL in neat plasma (EDTA), and 271 µg/mL in neat plasma Heparin.

  • The concentrations of Vitamin D Binding Protein were measured in duplicates, interpolated from the Vitamin D Binding Protein standard curves and corrected for sample dilution. All undiluted samples start at 1:500,000 in Sample Diluent NS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Vitamin D Binding Protein concentration was determined to be 5,280 ng /mL in neat milk, 288 ng/mL in neat saliva, and 56 ng /mL in neat urine.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Vitamin D Binding Protein concentration was determined to be 632 µg/mL with a range of 460 – 866 µg/mL.

Protocols

References

ab223586 has not yet been referenced specifically in any publications.

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