Human B7-H3 Quantikine ELISA Kit

(1 citations)   
  • 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 (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Urine (50 uL)
  • Sensitivity
    0.274 ng/mL
  • Assay Range
    0.8 - 50 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
  • Specificity
    Natural and recombinant human B7-H3
  • 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
QC47, Quantikine Immunoassay Control Set 725 for Human B7-H3 - Please Inquire
Product Summary
The Quantikine Human B7-H3 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human B7-H3 in cell culture supernates, serum, plasma, and urine. It contains NS0-expressed recombinant human B7-H3 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human B7-H3 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 human B7-H3.

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, Heparin Plasma, Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean5.891937.55.8817.635.1
Standard Deviation0.20.60.90.30.61.7
CV%3.43.32.55.13.55

EDTA Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean5.891937.55.8817.635.1
Standard Deviation0.20.60.90.30.61.7
CV%3.43.32.55.13.55

Recovery

The recovery of B7-H3 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 99-112
EDTA Plasma (n=4) 104 91-115
Heparin Plasma (n=4) 102 93-114
Serum (n=4) 103 94-117
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of B7-H3 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human B7-H3 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: B7-H3

Human B7-H3 was originally identified as a 316 amino acid (aa) type I membrane precursor protein with a putative 28 aa signal peptide, a 217 aa extracellular region containing one V-like and one C-like Ig domain, a transmembrane region, and a 45 aa cytoplasmic domain. Subsequently, a second dominantly expressed form of human B7-H3, containing a splice variation that duplicates the V-like and C-like Ig domains, was found. The 534 aa splice variant of B7-H3 has been referred to as B7-H3b, 4Ig-B7-H3, and B7-H3VCVC. RTPCR transcripts for both B7-H3 and 4Ig-B7-H3 have been found in all tissues except peripheral blood leukocytes. Southern blot analysis indicates that the 4Ig-B7-H3 isoform of B7-H3 is the predominate isoform expressed in human tissues.

In mouse, only a single form of B7-H3 containing one V-like and one C-like Ig domain was detected. Mouse and human B7-H3 share 87% aa sequence identity. B7-H3 has been shown to be expressed at very high levels in immature dendritic cells, at moderate levels on mature dendritic cells, LPS stimulated immature dendritic cells and LPS stimulated monocytes, and at low levels on resting monocytes. B7-H3 binds to activated T cells via an as yet unidentified receptor.

    • Long Name
      B7 Homolog 3
    • Entrez Gene IDs
      80381 (Human); 102657 (Mouse); 102131295 (Cynomolgus Monkey);
    • Alternate Names
      B7H3; B7-H3; B7H34Ig-B7-H3; B7-H3B7 homolog 3; CD276 antigen; CD276 molecule; CD276; Costimulatory molecule;
    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. 100 µL Assay Diluent
    4.   Add 100 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. 200 µL Conjugate
    9.   Add 200 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. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   Add 50 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. Soluble B7-H3 promotes the invasion and metastasis of pancreatic carcinoma cells through the TLR4/NF-?B pathway
      Authors: Chao Xie
      Sci Rep, 2016;6(0):27528.
      Species: Human
      Sample Type: Cell Culture Supernates
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 725 for Human B7-H3

    Ctrl QC47
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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