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    products/elisa/human-igg-elisa-kit-ab195215.pdf

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Immunology Immunoglobulins Heavy Chain IgG
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SimpleStep

Human IgG ELISA Kit (ab195215)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (5)Q&A (1)References (7)

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Other - Human IgG ELISA Kit (ab195215)
  • Example of Human IgG standard curve in Sample Diluent NS.
  • Raw data values for example of Human IgG standard curve in Sample Diluent NS
  • Example of Human IgG standard curve in 1X Cell Extraction Buffer PTR.
  • Raw data values for example of Human IgG standard curve in 1X Cell Extraction Buffer PTR
  • IgG levels in individual healthy donors.
  • Comparison of IgG levels in Human milk, urine and saliva.
  • IgG1-4 isotypes are detected by this kit
  • Linearity of dilution – native IgG in human serum, plasma (citrate, EDTA and Heparin), and culture media
  • Linearity of dilution – native IgG in human milk, urine, saliva and HLH extract

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 0.02 ng/ml
  • Range: 0.23 ng/ml - 15 ng/ml
  • Sample type: Cell culture supernatant, Cerebral Spinal Fluid, Cit plasma, EDTA Plasma, Hep Plasma, Milk, Saliva, Serum, Tissue Lysate, Urine
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

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Overview

  • Product name

    Human IgG ELISA Kit
    See all IgG kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 8 6.4%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 14.7%
  • Sample type

    Cell culture supernatant, Saliva, Milk, Urine, Serum, Hep Plasma, EDTA Plasma, Cit plasma, Tissue Lysate, Cerebral Spinal Fluid
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    0.02 ng/ml
  • Range

    0.23 ng/ml - 15 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Saliva 89 80% - 106%
    Milk 89 83% - 94%
    Urine 87 82% - 93%
    Serum 101 88% - 125%
    Cell culture media 107 96% - 115%
    Hep Plasma 100 100% - 100%
    EDTA Plasma 90.17 87% - 93%
    Cit plasma 100 98% - 102%
    Cerebral Spinal Fluid 98 88% - 105%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat, Sheep, Rabbit, Goat, Guinea pig, Cow, Dog
  • Product overview

    Human IgG ELISA kit (ab195215) is designed for the quantitative measurement of IgG protein in human serum, plasma, milk, urine, saliva, culture media and tissue extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human IgG with 20 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 SimpeStep ELISA® kits.


    CROSS REACTIVITY


    Human IgM, human IgA and human IgE were prepared at 10 ng/mL and 250 ng/mL in Sample Diluent NS and assayed for cross reactivity. No cross-reactivity was observed for IgM or IgE at either concentration with a mean OD deviation from background of -0.01. No cross-reactivity was observed for IgA at 10 ng/mL and only 0.4% cross-reactivity at 250 ng/mL of IgA.

    INTERFERENCE 


    Purified human IgG was assayed at 5 ng/mL in the presence and absence of 250 ng/mL of human IgM, human IgA and human IgE to determine interference. After background subtraction, recovery of human IgG was observed at a mean of 98% with a standard deviation of 0.06.

    SPECIES REACTIVITY 


    This kit recognizes human IgG protein.
    Other species reactivity was determined by measuring 10 ng/mL of purified IgG from various species, interpolating the protein concentrations from the human standard curve, and expressing the interpolated concentrations as a percentage of the protein concentration of human IgG assayed at the same concentration.
    Reactive species: Rhesus Monkey
    Reactivity < 3% was determined for the following species: Mouse, Rat, Rabbit, Dog, Goat, Sheep, Cow, Guinea Pig


    CALIBRATION 


    ab195215 (IgG) - This immunoassy is calibrated against a highly purified human IgG. The NIBSC/WHO unclassified purified human IgG/IgM/IgG preperation 67/086. was evaluatied in this kit.
    The dose response curve of the unclassified standard parallels the SimpleStep standard curve. To convert sample values obtained with the SimpleStep IgG kit to approximate NIBSC International units, use the equation below.
    NIBSC 67/086 approximate value (IU/mL) = 0.000017 (1.7e-5 IU) x SimpleStep IgG value (ng/mL)

  • Notes

    Immunoglubulin G (IgG) is a glycoprotein molecule which belongs to the immunoglobulin family of proteins known as antibodies. Immunoglobulins are the key component of humoral immunity. IgG is a monomeric immunoglobulin, built of two heavy chains gamma and two light chains. The heavy chains are linked to each other and to the light chain by disulfide bonds. Each molecule has two antigen binding sites. While the amino-terminal portions that exhibit highly variable amino-acid composition are involved in antigen binding, the C terminal constant parts are involved in complement binding, placental passage and binding to cell membrane. This is the most abundant immunoglobulin and is approximately equally distributed in blood and in tissue liquids, constituting 75% of serum immunoglobulins in humans. This is the only isotype that can pass through the human placenta, thereby providing protection to the fetus in its first weeks of life before its own immune system has developed. It can bind to many kinds of pathogens, for example viruses, bacteria, and fungi, and protects the body against them by complement activation (classic pathway), opsonization for phagocytosis and neutralisation of their toxins. There are 4 subclasses: IgG1 (66%), IgG2 (23%), IgG3 (7%) and IgG4 (4%).

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human IgG Capture Antibody 1 x 600µl
    10X Human IgG Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CP 1 x 6ml
    Human IgG Lyophilized Purified Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

    • Immunology
    • Immunoglobulins
    • Heavy Chain
    • IgG
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Immunoglobulins ELISA kits
  • Cellular localization

    Secreted
  • Alternative names

    • Ig gamma 1 chain C region
    • Ig gamma 2 chain C region
    • Ig gamma 3 chain C region
    • Ig gamma 4 chain C region
    • IgG
    • IGHG1
    • IGHG2
    • IGHG3
    • IGHG4
    • Immunoglobin heavy constant gamma 1
    • Immunoglobulin G
    see all
  • Database links

    • Entrez Gene: 3500 Human
    • Entrez Gene: 3501 Human
    • Entrez Gene: 3502 Human
    • Entrez Gene: 3503 Human
    • SwissProt: P01857 Human
    • SwissProt: P01859 Human
    • SwissProt: P01860 Human
    • SwissProt: P01861 Human

    Images

    • Other - Human IgG ELISA Kit (ab195215)
      Other - Human IgG ELISA Kit (ab195215)

      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 Human IgG standard curve in Sample Diluent NS.
      Example of Human IgG standard curve in Sample Diluent NS.

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

    • Raw data values for example of Human IgG standard curve in Sample Diluent NS
      Raw data values for example of Human IgG standard curve in Sample Diluent NS

      Raw data values are shown in the table

    • Example of Human IgG standard curve in 1X Cell Extraction Buffer PTR.
      Example of Human IgG standard curve in 1X Cell Extraction Buffer PTR.

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

    • Raw data values for example of Human IgG standard curve in 1X Cell Extraction Buffer PTR
      Raw data values for example of Human IgG standard curve in 1X Cell Extraction Buffer PTR

      Raw data values are shown in the table

    • IgG levels in individual healthy donors.
      IgG levels in individual healthy donors.

      Ten individual healthy donors were evaluated for the presence of IgG in serum using this assay. Results were interpolated from the standard curve in Sample Diluent NS and corrected for sample dilution (1:5x106). The mean level of IgG was found at 12.5 mg/mL with a range of 9 – 18.4mg/mL.

    • Comparison of IgG levels in Human milk, urine and saliva.
      Comparison of IgG levels in Human milk, urine and saliva.

      Bodily fluids from 3 different donors were evaluated for the presence of IgG using this assay. Results were interpolated from the standard curve in sample diluent NS and corrected for sample dilution (1:2.5x104). The mean levels in Milk were found at 20.7 µg/mL, in Urine at 0.8 µg/mL and in Saliva at 11.1 µg/mL.

    • IgG1-4 isotypes are detected by this kit
      IgG1-4 isotypes are detected by this kit

      Human IgG1, IgG2, IgG3 and IgG4 were tested at 5ng/mL

    • Linearity of dilution – native IgG in human serum, plasma (citrate, EDTA and Heparin), and culture media
      Linearity of dilution – native IgG in human serum, plasma (citrate, EDTA and Heparin), and culture media

      Native IgG was measured in the human serum, plasma citrate, plasma EDTA, plasma heparin and culture media in a 2-fold dilution series. Sample dilutions were made in Sample Diluent NS for all samples, purified IgG was spiked into culture media and diluted in a 2-fold dilution series in Sample Diluent NS

    • Linearity of dilution – native IgG in human milk, urine, saliva and HLH extract
      Linearity of dilution – native IgG in human milk, urine, saliva and HLH extract

      Native IgG was measured in milk, saliva, urine and liver homogenate (HLH) in a 2-fold dilution series. Sample dilutions were made in Sample Diluent NS for all samples except for HLH, which was carried out in 1X cell extraction buffer PTR.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (7)

    Publishing research using ab195215? Please let us know so that we can cite the reference in this datasheet.

    ab195215 has been referenced in 7 publications.

    • Smith P  et al. Characterization of ultrasound-mediated delivery of trastuzumab to normal and pathologic spinal cord tissue. Sci Rep 11:4412 (2021). PubMed: 33627726
    • Dong Q  et al. Astragalus-mediated stimulation on antigen-presenting cells could result in higher IL-21 production from CXCR5+ Tfh-like cells and better IL-21-mediated effector functions. Hum Immunol 82:429-437 (2021). PubMed: 33875296
    • Cuhadar U  et al. Autoantibodies produce pain in complex regional pain syndrome by sensitizing nociceptors. Pain 160:2855-2865 (2019). PubMed: 31343542
    • Vatine GD  et al. Human iPSC-Derived Blood-Brain Barrier Chips Enable Disease Modeling and Personalized Medicine Applications. Cell Stem Cell 24:995-1005.e6 (2019). PubMed: 31173718
    • Adler AS  et al. A natively paired antibody library yields drug leads with higher sensitivity and specificity than a randomly paired antibody library. MAbs 10:431-443 (2018). ELISA . PubMed: 29376776
    • Akyol GY  et al. IVIG activates Fc?RIIB-SHIP1-PIP3 Pathway to stabilize mast cells and suppress inflammation after ICH in mice. Sci Rep 7:15583 (2017). PubMed: 29138419
    • Wang N  et al. Herbal Medicine Cordyceps sinensis Improves Health-Related Quality of Life in Moderate-to-Severe Asthma. Evid Based Complement Alternat Med 2016:6134593 (2016). ELISA ; Human . PubMed: 28050193

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-6 of 6 Abreviews or Q&A

    good elisa

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    We obtain reliable and reproducible results with this Elisa.
    The protocol booklet is well explained, easy to understand and easy to perform.
    We used this kit for serum samples and cell culture supernatants.
    The kit is easy to use and quick compared to other Sandwich Elisa.
    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.

    Abcam user community

    Verified customer

    Submitted Jul 05 2022

    Quantification of Total IgG in Human Fecal Samples

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    We have used Abcam's Human IgG ELISA kit to examine total IgG level within our human fecal samples. We have processed the fecal samples according to our standard protocol and used the extraction for the experiment.

    Samples from two different volunteer subjects have been selected for this trial assay. The samples from each subject had been collected at four different time points during the clinical trial.We have tested four different sample dilutions (1:10, 1:100, 1:500, 1:1000) among these samples to find the ideal level of detection.

    According to our attached results, dilutions between 1:100 and 1:500 may be ideal for measuring total IgG levels in human fecal samples using this kit.
    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.

    Aaron Kim

    Verified customer

    Submitted Aug 14 2020

    Quantification of Total IgG in Human Fecal Samples

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    We have used Abcam's Human IgG ELISA kit to examine total IgG level within our human fecal samples. We have processed the fecal samples according to our standard protocol and used the extraction for the experiment.

    Samples from two different volunteer subjects have been selected for this trial assay. The samples from each subject had been collected at four different time points during the clinical trial.We have tested four different sample dilutions (1:10, 1:100, 1:500, 1:1000) among these samples to find the ideal level of detection.

    According to our attached results, dilutions between 1:100 and 1:500 may be ideal for measuring total IgG levels in human fecal samples using this kit.
    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.

    Aaron Kim

    Verified customer

    Submitted Aug 13 2020

    reaction time

    Below average Good 4/5 (Ease of Use)
    Abreviews
    Abreviews
    It is better to observe the plate to be able to stop the reaction early if necessary. In our case (we stopped after 5 minutes) already 2 values of the calibration curve were no longer measurable (OD too high)
    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.

    Mrs. Heidrun Jäger

    Verified customer

    Submitted Jun 14 2019

    works very well as explained

    Good Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    used it to quantify IgGs in Heparin plasma
    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.

    Abcam user community

    Verified customer

    Submitted May 14 2018

    Question

    Can you tell me if the referenced kit has any biotin-streptavidin components to it?

    Read More

    Abcam community

    Verified customer

    Asked on Jun 23 2016

    Answer

    No, there are no biotin-streptavidin components in this human IgG ELISA, product ab195215.

    Read More

    Tom Ruyle

    Abcam Scientific Support

    Answered on Jun 23 2016

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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