Overview

  • Product name
    Human Apolipoprotein A-IV ELISA Kit, Fluorescent
    See all APOA4/Apo-AIV kits
  • Detection method
    Fluorescent
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Human serum 8 3.8%
    Inter-assay
    Sample n Mean SD CV%
    Human serum 3 6.4%
  • Sample type
    Cell culture supernatant, Saliva, Urine, Serum, Tissue Extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    10.8 pg/ml
  • Range
    39.1 pg/ml - 160000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Saliva 78 76% - 79%
    Urine 97 95% - 98%
    Serum 98 90% - 101%
    Tissue Extracts 103 102% - 105%
    Cell culture media 103 102% - 103%
    Heparin Plasma 98 95% - 101%
    EDTA Plasma 104 102% - 107%
    Citrate Plasma 91 90% - 94%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat, Cow
  • Product overview

    Apolipoprotein A-IV in vitro CatchPoint® SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Apolipoprotein A-IV protein in human serum, plasma, saliva, urine, cell culture supernatants, and tissue extract samples.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.


    The CatchPoint® 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. CatchPoint® HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.

  • Notes

    Human Apolipoprotein A-IV is a 45kDa glycoprotein with activities involving lipid transport. Specifically, Apolipoprotein A-IV is a component of high density lipoprotein (HDL) and chylomicrons. Its primarily synthesized is the intestine in association with lymph chylomicron particles. Although its precise function is not known, Apolipoprotein A-IV is a potent activator of lecithin-cholesterol acyltransferase (LCAT) in vitro. Human Apolipoprotein A-IV consists of a 20 amino acid (aa) signal peptide and 376 aa mature chain. Human Apolipoprotein A-IV is 62% identical to both mouse and rat Apolipoprotein A-IV.

  • 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 ab229428 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
  • The Apolipoprotein A-IV standard curve was prepared as described in Section 10. Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Apolipoprotein A-IV were measured in duplicates, interpolated from the Apolipoprotein A-IV standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:32,000, plasma (citrate) 1:16,000, plasma (EDTA) 1:16,000, and plasma (heparin) 1:16000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Apolipoprotein A-IV concentration was determined to be 71.65 µg/mL in neat serum, 48.04 µg/mL in neat plasma (citrate), 42.94 µg/mL in neat plasma (EDTA) and 51.18 µg/mL in neat plasma (heparin).

  • The concentrations of Apolipoprotein A-IV were measured in duplicate and interpolated from the Apolipoprotein A-IV standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Apolipoprotein A-IV concentration was determined to be 4,829 pg/mL in placenta extract and 17,016 pg/mL in liver homogenate extract.

  • The concentrations of Apolipoprotein A-IV were measured in duplicates, interpolated from the Apolipoprotein A-IV standard curves and corrected for sample dilution. Undiluted samples are as follows: saliva 6.25% and urine 12.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Apolipoprotein A-IV concentration was determined to be 177,941 pg/mL in neat saliva (spiked) and 80,680 pg/mL in neat urine (spiked).

  • Saliva was analyzed at 6.25% and urine at 12.5%. The concentrations of Apolipoprotein A-IV were measured in duplicates, interpolated from the Apolipoprotein A-IV standard curves and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Apolipoprotein A-IV concentration was determined to be 25,370 pg/mL in neat saliva (native) and 3903 pg/mL in neat urine (native).

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean human Apolipoprotein A-IV concentration was determined to be 192.8 µg/mL with a range of 9.2 – 418.0 µg/mL.

Protocols

References

ab229428 has not yet been referenced specifically in any publications.

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