Mouse alpha-Fetoprotein/AFP Quantikine ELISA Kit

(1 citations)
(1 Review)
  
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
  • Sensitivity
    0.029 ng/mL
  • Assay Range
    0.2 - 10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant mouse AFP.
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine Mouse alpha -Fetoprotein/AFP Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse AFP levels in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse AFP and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse AFP accurately. Results obtained using natural mouse AFP showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse AFP.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean0.4041.123.30.4571.23.38
Standard Deviation0.0130.0260.0910.0310.0660.158
CV%3.22.32.86.85.54.7

Recovery

The recovery of mouse AFP spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 98 93-105
Linearity
To assess the linearity of the assay, samples containing high concentrations of mouse AFP were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
Mouse alpha-Fetoprotein/AFP Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: alpha-Fetoprotein/AFP
alpha-Fetoprotein (AFP), is albuminoid superfamily protein that is synthesized in the fetus primarily by the liver, yolk sac, and tissues of gastrointestinal origin. It is one of the earliest markers of the hepatocyte lineage. AFP acts as a carrier protein for steroids, bilirubin, fatty acids, retinoids, and flavonoids. In addition, it can exert immunosuppressive activity, regulate cell proliferation and apoptosis, initiate intracellular signaling, and contribute to cell invasion. Altered levels of both fetal and maternal AFP have been associated with hypothyroidism, autoimmune disorders, and heart defects. Low maternal serum AFP levels are associated with a higher incidence of Down syndrome, whereas higher levels are associated with spina bifida and anencephaly. Elevated AFP levels are also coincident with liver, stomach, and germ cell cancers.
    • Entrez Gene IDs
      174 (Human); 11576 (Mouse); 24177 (Rat);
    • Alternate Names
      AFP; Alpha-1-fetoprotein; alpha-fetoglobulin; alpha-Fetoprotein; DSCAM2; FETA; HP; HPAFP;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 50 µL Assay Diluent
    4.   Add 50 µL of Assay Diluent to each well.

    5. 50 µL Standard, Control, or Sample
    6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 100 µL Conjugate
    9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
    10.   Aspirate and wash 4 times.

    11. 100 µL Substrate Solution
    12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    13. 100 µL Stop Solution
    14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    Citations:

    R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

    1 Citations: Showing 1 - 1

    1. Adult mouse model of early hepatocellular carcinoma promoted by alcoholic liver disease
      Authors: A Ambade, A Satishchan, B Gyongyosi, P Lowe, G Szabo
      World J Gastroenterol, 2016;22(16):4091-108.
      Species: Mouse
      Sample Type: Serum
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    Average Rating: 5 (Based on 1 review)

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     Mouse alpha-Fetoprotein/AFP Quantikine ELISA Kit MAFP00
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     Mouse alpha-Fetoprotein/AFP Quantikine ELISA Kit MAFP00
    : Mouse alpha-Fetoprotein/AFP Quantikine ELISA Kit [MAFP00]

    Summary

    Sample TestedMouse sera diluted 120x in Calibrator Diluent RD5-26 (diluted 1:4)

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