Overview

  • Product name
    Human Retinol Binding Protein 4 ELISA Kit
    See all Retinol binding Protein 4 kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Serum 5 5.1%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 8.9%
  • Sample type
    Cell culture supernatant, Saliva, Milk, Urine, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    2.6 pg/ml
  • Range
    15.6 pg/ml - 1000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Saliva 104 99% - 108%
    Milk 103 100% - 108%
    Urine 110 106% - 113%
    Serum 109 104% - 111%
    Cell culture media 101 99% - 102%
    Heparin Plasma 103 91% - 110%
    EDTA Plasma 108 106% - 110%
    Citrate Plasma 109 106% - 114%
    serum free media 103 93% - 113%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Product overview

    RBP4 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of RBP4 protein in human serum, plasma, urine, saliva, milk, and cell culture supernatant. 


    Human RBP4 SimpleStep ELISA® kit (ab196264) has been redeveloped with new capture and detector antibodies. While this does not affect the specificity of the results, it may result in increased sensitivity and will improve the consistency in the performance of the kit.


    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.


    Retinol-binding protein 4 (RBP4) is a circulating protein that binds to and transports retinol (vitamin A) in serum. The RBP4-retinol complex further interacts with prealbumin (also known as transthyretin) which increases its half-life in circulation. RBP4 is produced by hepatocytes and adipocytes. Loss of functional RBP4 protein results in serum retinol deficiency which can impact vision.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate (12 x 8 well strips)

Properties

Applications

Our Abpromise guarantee covers the use of ab196264 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 Protocol Summary
  • Standard curve comparison between Human RBP4 SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.

  • The RBP4 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 RBP4 were measured in duplicates, interpolated from the RBP4 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:400,000, plasma (citrate) 1:200,000, plasma (EDTA) 1: 100,000, and plasma (heparin) 1:200,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean RBP4 concentration was determined to be 22.2 µg/mL in serum, 21.9 µg/mL in plasma (citrate), 15.1 µg/mL in plasma (EDTA) and 52.9 µg/mL in plasma (heparin).

  • The concentrations of RBP4 were measured in duplicates, interpolated from the RBP4 standard curves and corrected for sample dilution. Undiluted samples are as follows:  saliva 1:80, urine 1: 160, milk 1:1000, and HEPG2 cell culture supernatant 1:1000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean RBP4 concentration was determined to be 16.2 ng/mL in saliva, 66.3 ng/mL in urine, 111 ng/mL in milk and 1071 ng/mL in HEPG2 cell culture supernatant.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean RBP4 concentration was determined to be 44 µg/mL with a range of 28 – 67 µg/mL.

  • Bovine reactivity was seen at serum levels greater than 0.03%.

Protocols

References

ab196264 has not yet been referenced specifically in any publications.

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