Human CD23/Fc epsilon RII Quantikine ELISA Kit

  
  • 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 (30 uL), EDTA Plasma (30 uL), Heparin Plasma (30 uL), Urine (30 uL)
  • Sensitivity
    5.1 pg/mL
  • Assay Range
    31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
  • Specificity
    Natural and recombinant human CD23. This assay recognizes the 25 kDa fragment (aa 150-321) and the full length extracellular domain (aa 48-321) of human CD23/Fc epsilon RII.
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC24, Quantikine Immunoassay Control Group 9 - Please Inquire
Product Summary
The Quantikine Human CD23 immunoassay is a 4.5 hour solid phase ELISA designed to measure human soluble CD23 in cell culture supernates, serum, plasma, and urine. It contains NS0-expressed recombinant human CD23 (extracellular domain) and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human CD23 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring soluble CD23.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean158283576151274533
Standard Deviation13.517.840.813.722.440.2
CV%8.56.37.19.18.27.5

Recovery

The recovery of CD23 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 105 94-114
EDTA Plasma (n=4) 99 89-105
Heparin Plasma (n=4) 98 90-112
Serum (n=4) 94 85-108
Urine (n=4) 100 85-113
Linearity
To assess the linearity of the assay, samples containing high concentrations of CD23 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human CD23/Fc epsilon RII Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: CD23/Fc epsilon RII
CD23, also known as Fc epsilon RII, is a type 2 transmembrane C-type lectin that binds IgE, CD21, CD11b and CD11c. It is expressed on a variety of hematopoietic cells and functions in allergic immune responses.
    • Long Name
      Fc epsilon Receptor II
    • Entrez Gene IDs
      2208 (Human); 14128 (Mouse); 171075 (Rat);
    • Alternate Names
      BLAST-2; CD23; CD23A; CD23CD23 antigen; CLEC4J; CLEC4JC-type lectin domain family 4 member J; C-type lectin domain family 4, member J; Fc epsilon RII; Fc fragment of IgE, low affinity II, receptor for (CD23); FCE2Fc fragment of IgE, low affinity II, receptor for (CD23A); fc-epsilon-RII; FCER2; Fcer2a; FceRII; IGEBF; Immunoglobulin E-binding factor; low affinity immunoglobulin epsilon Fc receptor; Ly-42; Lymphocyte IgE receptor;
    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 3 hours.
    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 1 hour.
    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. 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.
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Group 9

    Ctrl QC24
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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