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RecombinantSimpleStep

Rat Retinol Binding Protein 4 ELISA Kit (ab203362)

Reviews (1) Submit a question References (2)
Other - Rat Retinol Binding Protein 4 ELISA Kit (ab203362)
  • Example of RBP4 standard curve prepared in Sample Diluent NS.
  • Interpolated concentrations of RBP4 in rat plasma (citrate) and serum.
  • Sandwich ELISA - Rat Retinol Binding Protein 4 ELISA Kit (ab203362)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 3.07 pg/ml
  • Range: 13.3 pg/ml - 850 pg/ml
  • Sample type: Cell culture supernatant, Plasma, Serum, Urine
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Rat

Overview

  • Product name

    Rat Retinol Binding Protein 4 ELISA Kit
    See all RBP4 kits

  • Detection method

    Colorimetric

  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 5 3.3%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 2.7%

  • Sample type

    Cell culture supernatant, Urine, Serum, Plasma

  • Assay type

    Sandwich (quantitative)

  • Sensitivity

    3.07 pg/ml

  • Range

    13.3 pg/ml - 850 pg/ml

  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 116 109% - 126%
    Urine 104 103% - 105%
    Serum 113 107% - 116%
    Cit plasma 100 91% - 106%

  • Assay time

    1h 30m

  • Assay duration

    One step assay

  • Species reactivity

    Reacts with: Rat
    Does not react with: Sheep, Goat, Cow, Pig

  • Product overview

    Rat Retinol Binding Protein 4 ELISA Kit (ab203362) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Retinol Binding Protein 4 protein in cell culture supernatant, serum, urine, and plasma. It uses our proprietary SimpleStep ELISA® technology. Quantitate Rat Retinol Binding Protein 4 with 3.07 pg/ml sensitivity.


    SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:


            - Single-wash protocol reduces assay time to 90 minutes or less
            - High sensitivity, specificity and reproducibility from superior antibodies
            - Fully validated in biological samples
            - 96-wells plate breakable into 12 x 8 wells strips


    A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

  • Notes

    Retinol-Binding Protein 4 (RBP4) is a small, highly conserved member of the lipocalin superfamily involved in the transportation of retinol (Vitamin A) in blood. Synthesized in hepatocytes and adipocytes, RBP4 is also an adipokine involved in insulin resistance in the AG4KO mouse model. Elevated serum levels of RBP4 are seen in obesity and Type 2 diabetes.

  • Platform

    Microplate (12 x 8 well strips)

Properties

Images

  • Other - Rat Retinol Binding Protein 4 ELISA Kit (ab203362)
    Other - Rat Retinol Binding Protein 4 ELISA Kit (ab203362)

    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.

     

  • Example of RBP4 standard curve prepared in Sample Diluent NS.
    Example of RBP4 standard curve prepared in Sample Diluent NS.

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

  • Interpolated concentrations of RBP4 in rat plasma (citrate) and serum.
    Interpolated concentrations of RBP4 in rat plasma (citrate) and serum.

    The concentrations of RBP4 were measured in duplicate and interpolated from the RBP4 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

  • Sandwich ELISA - Rat Retinol Binding Protein 4 ELISA Kit (ab203362)
    Sandwich ELISA - Rat Retinol Binding Protein 4 ELISA Kit (ab203362)
    To learn more about the advantages of recombinant antibodies see here.

References (2)

Publishing research using ab203362? Please let us know so that we can cite the reference in this datasheet.

ab203362 has been referenced in 2 publications.

  • Hoffman JF  et al. Effect of embedded metal fragments on urinary metal levels and kidney biomarkers in the Sprague-Dawley rat. Toxicol Rep 8:463-480 (2021). PubMed: 33717999
  • Sancho-Martínez SM  et al. Impaired Tubular Reabsorption Is the Main Mechanism Explaining Increases in Urinary NGAL Excretion Following Acute Kidney Injury in Rats. Toxicol Sci 175:75-86 (2020). PubMed: 32110797

Customer reviews and Q&As

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Submit a review Submit a question

Rat RBP4 ELISA kit unlikely to have adequate accuracy through cross-reactivity in Mongolian gerbil serum samples

 Inconclusive Below average 2/5 (Ease of Use)
Abreviews
abreview image
I needed to assay retinol binding protein 4 in serum samples from Meriones unguiculatus however, commercial antibodies are not available. Old research indicates that there can be crossreactivity between rat anti-RBP4 antiserum and gerbil RBP4, and the Abcam customer service rep I discussed this with suggested that due to 98% identity in sequence, the kit may work in this species, so I enrolled in the Abtrial program to test this kit. The kit requires (in rat samples) that serum is diluted 1:64,000-1:640,000 times, so I calculated a dilution factor of 1:100,000x based on my serum retinol concentrations (HPLC determined) in the samples to put RBP concentrations near the middle of the external standardization curve. Obviously, a 5-step serial dilution prior to analysis in 66 samples is not ideal, but was performed using an electronic multichannel pipette for accuracy and efficiency in 96-well plates as 5 x 15 uL sample:135 uL diluent (this allows all dilutions to occur in the provided 50 mL of diluent "NS") with mixing in between steps.

All steps were performed according to the booklet, with samples run at the same time as the standard curve. All samples were within the standard curve, which was determined by 4-factor logistic regression as recommended. One low concentration point did not fit the curve very well but the samples were not near this concentration. We were planning on running a second kit from the same lot if this first one appeared to work as a duplicate to ensure that all samples were run on each plate.

I believe that this kit did not work in our samples as the results do not correlate at all (R-squared of ~0) with serum retinol concentrations in fasted gerbils, a well-known correlation in other rodents and mammals due to the toxicity of free retinol in serum. 11 of the 66 samples indicated that there was less RBP than retinol on a molar basis, half of which with than 50% of retinol, which would be extremely harmful to the animal if true.

It is interesting that the values did fall along the standard curve, indicating that the dilution was adequate, and were varied over a somewhat plausible range ignoring the relation to serum retinol.

I have attached images of the standard curve, a plot of the ELISA values vs serum retinol, and an image of the plate just prior to a final mix and reading at 450nm.

Unfortunately, recombinant gerbil RBP4 is not commercially available, so we could not definitively rule out other sources of error on this experiment.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted Jun 20 2018

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