Thromboxane B2 Parameter Assay Kit

(4 citations)   
  • Format
    96-well strip plate
  • Assay Length
    3.5 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
    0.31 ng/mL
  • Assay Range
    0.3 - 20 ng/mL (Cell Culture Supernates, Serum, Plasma, EDTA Plasma, Heparin Plasma, Urine)
  • Specificity
    Measures TXB2 levels in various samples
  • Cross-reactivity
    Cross-reactivity observed with 1 or more available related molecules.Cross-species reactivity not tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC179, Parameter Immunoassay Control Set 931 for Thromboxane B2 - Please Inquire
Product Summary
The Parameter TXB2 Immunoassay is a 3.5 hour competitive enzyme immunoassay designed to measure TXB2 in cell culture supernates, serum, plasma, and urine.

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
Mean3.876.9513.23.867.0514.4
Standard Deviation0.230.380.510.340.450.74
CV%5.95.53.98.96.45.1

Recovery

The recovery of TXB2 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 98 88-110
EDTA Plasma (n=4) 94 86-105
Heparin Plasma (n=4) 97 87-107
Serum (n=4) 98 86-111
Urine (n=4) 98 84-111
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of TXB2 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Thromboxane B2 Parameter Assay Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Thromboxane B2
Thromboxane A2 (TXA2), is an eicosanoid involved in platelet aggregation, vasoconstriction, and reproductive functions. However, as TXA2 is rapidly degraded in vivo, TXA2 production is typically examined by assaying the more stable Thromboxane B2 (TXB2) or 11-dehydro-TXB2 (11-d-TXB2) derivatives. Levels of these metabolites are helpful markers in the study of diseases including liver cirrhosis, cystic fibrosis, mastocytosis, systemic lupus erythematosus, thrombosis diseases, and others.
    • Long Name
      Thromboxane B2
    • Alternate Names
      Thromboxane B2; TXB2;
    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. Calibrator Diluent
    4.   Add 150 µL of Calibrator Diluent to the non-specific binding (NSB) wells.
    5.   Add 100 µL of Calibrator Diluent to the zero standard (B0) wells.

    6. 100 µL Standard, Control, or Sample
    7.   Add 100 µL of Standard, control, or sample to the remaining wells.

    8. 50 µL Primary Antibody Solution
    9.   Add 50 µL of Primary Antibody Solution to each well (except the NSB wells).
    10.   Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker. Do not wash the plate.

    11. 50 µL Conjugate
    12.   Add 50 µL of Conjugate to each well. Cover with a plate sealer, and incubate at room temperature for 1 hour on the shaker.
    13.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    14. 200 µL Substrate Solution
    15. Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    16. 100 µL Stop Solution
    17. 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.

    4 Citations: Showing 1 - 4
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    Species
    Sample Type
    1. Acetylsalicylic acid differentially limits the activation and expression of cell death markers in human platelets exposed to Staphylococcus aureus strains
      Authors: A Chabert, P Damien, PO Verhoeven, F Grattard, P Berthelot, F Zeni, L Panicot-Du, S Robert, F Dignat-Geo, MA Eyraud, B Pozzetto, B Payrastre, F Cognasse, O Garraud, H Hamzeh-Cog
      Sci Rep, 2017;7(1):5610.
      Species: Human
      Sample Type: Cell Culture Supernates
    2. Directed transport of neutrophil-derived extracellular vesicles enables platelet-mediated innate immune response
      Nat Commun, 2016;7(0):13464.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    3. Anti-tumor and Anti-angiogenic Effects of Aspirin-PC in Ovarian Cancer
      Mol Cancer Ther, 2016;0(0):.
      Species: Mouse
      Sample Type: Serum
    4. In vivo modulation of the inflammatory response by nonsteroidal antiinflammatory drug-related compounds that trigger L-selectin shedding.
      Authors: Herrera-Garcia A, Dominguez-Luis M, Arce-Franco M, Lopez-Fernandez J, Feria M, Barreiro O, Sanchez-Madrid F, Diaz-Gonzalez F
      Eur J Immunol, 2013;43(1):55-64.
      Species: Mouse
      Sample Type: Cell Culture Supernates
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    Parameter Immunoassay Control Set 931 for Thromboxane B2

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