IISBR
  • Home
  • Meet Our Team
    • IISBR Team Members
    • Postdoctoral Researchers
    • Faculty Affiliates
  • Assay Services
    • Assay Service Menu
    • Sample Collection and Shipping
  • IISBR Research
    • IISBR Google Scholar
    • IISBR Publications
  • Courses and Trainings
    • Undergraduate and Graduate Courses
    • Spit Camp I
    • Spit Camp II
  • Events
  • Contact Us
  • Forms
    • Assay Quote
    • Spit Camp I Pre-Registration
    • Spit Camp II Pre-Registration

Immunoglobulin G (IgG)

    Volume Requirements

    10 ul saliva

    Diluted 1:2500 prior to testing

    (100ul required for testing to account for pipetting needs and sample quality)

    Assay Range

    0.3125 - 20 ng/mL

    Assay Sensitivity

    1.33 µg/mL

    Compatible Collection Methods

    Passive Drool

    Swab

    Special Considerations

    Flow Rate Dependent (time the collection and weigh samples in lab)*

    *Not important if IgG only being used to qualify samples as to whether they have sufficient antibody levels

    Methodology

    Saliva is tested at IISBR using a commercially-available enzyme-linked immunosorbent assay kit (ELISA) from Salimetrics following manufacturer’s protocol without modification. This test employs a 96-well microtitre plate coated with a monoclonal antibody (Ab) to IgG. IgG in standards, controls and unknown samples bind to available antibody binding sites on the plate. A secondary antibody linked to horseradish peroxidase is added and binds to the bound IgG from standards, controls and samples. Bound peroxidase is measured by the reaction of the peroxidase enzyme on the substrate tetramethylbenzidine (TMB) producing a blue color. The reaction is stopped and the resultant yellow color is read at 450nm on a spectrophotometer (PowerWave HT, BioTek Instruments). The amount of IgG peroxidase detected, as measured by the intensity of color, is directly proportional to the amount of IgG present.

    Briefly, 100 µL of standard, controls, and unknown, diluted saliva are pipetted onto 96-well microtitre plates and incubated for 2 hours shaking at room temperature. Plates are washed 4 times followed by addition the prepared IgG enzyme conjugated to HRP. Plate is incubated for 2 hours shaking at room temperature, followed by the addition of a TMB substrate. Plate is incubated shaking at room temperature in the dark for 30 minutes. Following incubation, the reaction is stopped with 2M Sulfuric Acid (NH2SO4)and optical density (OD) read at 450nm with a 620nm correction filter. Concentrations of controls and samples are calculated from a standard curve generated using a 4-parameter non-linear regression curve fit (Gen5, BioTek).

    • Request a Quote

      Quote Form

    UCI School of Social Ecology
    Social Ecology I
    Irvine, CA 92697-7050
    www.uci.edu
    www.socialecology.uci.edu

    UCI Program in Public Health
    UCI Health Sciences Complex
    856 Health Sciences Quad
    Irvine, CA 92697-3957
    www.uci.edu www.publichealth.uci.edu

    Log-In

    © 2023 UC Regents