Overview

  • Product name

    Urinary Prostacyclin ELISA kit
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Low 24 23pg/ml 9.1%
    Medium 24 66pg/ml 3.4%
    High 24 200pg/ml 2.9%
    Inter-assay
    Sample n Mean SD CV%
    Low 8 23pg/ml 9.6%
    Medium 8 72pg/ml 5.4%
    High 8 223pg/ml 3.4%
  • Sample type

    Cell culture supernatant, Urine, Tissue Culture Media
  • Assay type

    Competitive
  • Sensitivity

    6.58 pg/ml
  • Range

    7.81 pg/ml - 2000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 101.9 % - %
    Tissue Culture Media 109.2 % - %

  • Assay time

    3h 0m
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
    Predicted to work with: a wide range of other species
  • Product overview

    Abcam’s Human Urinary Prostacyclin in vitro competitive ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the accurate quantitative measurement of Prostacyclin breakdown products in urine. The kit measures the two major breakdown products of Prostacyclin in urine, 6-keto-Prostaglandin F1 alpha (6k-PGF1 alpha) and 2,3-dinor-6-keto-Prostaglandin F1 alpha (2,3d-6k-PGF1 alpha).

    A donkey anti-sheep IgG antibody has been precoated onto 96-well plates. Standards or test samples are added to the wells, along with an alkaline phosphatase (AP) conjugated- Urinary Prostacyclin antigen and a polyclonal sheep antibody specific to Urinary Prostacyclin. After incubation the excess reagents are washed away. pNpp substrate is added and after a short incubation the enzyme reaction is stopped and the yellow color generated is read at 405 nm. The intensity of the yellow coloration is inversely proportional to the amount of Urinary Prostacyclin captured in the plate.

  • Notes

    Prostacyclin (PGI2) is involved in platelet aggregation, vasoconstriction, and reproductive functions. PGI2 has a half life of 60 minutes in plasma but only 2 to 3 minutes in buffer. The production of PGI2 is typically monitored by measurement of 6k-PGF and 2,3d-6k- PGF. 6-keto- PGF is produced by the non-enzymatic hydration of PGI2, and is further metabolized in urine to the 2,3-dinor derivative. A number of pharmaceuticals alter and/or inhibit the synthesis of PGI2. Methods to measure PGI2 in urine typically involve HPLC, gas chromatography/mass spectrometry, or radioimmunoassay and enzyme immunoassay for 6k- PGF.

    Cross Reactivity

    The cross reactivity for a number of related eicosanoid compounds was determined.

    CompoundCross Reactivity
    2,3-dinor-6-keto-PGF100%
    6-keto-PGF89.14%
    PGF13.21%
    PGF4.99%
    13,14 dihydro-PGF2.55%
    15-keto-PGF0.32% 
    PGE10.08%
    PGD20.03%
    6,15-diketo-13,14 dihydro-PGF0.03%
    PGB10.01%
    PGE20.01%
    PGA20.01%
    TXB20.01%
  • Tested applications

    Suitable for: Competitive ELISAmore details
  • Platform

    Microplate

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab133032 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Competitive ELISA Use at an assay dependent concentration.

Images

  • Representative Standard Curve using ab133032

Protocols

References

This product has been referenced in:

  • Umeda S  et al. Enhanced Pulmonary Vascular and Alveolar Development via Prenatal Administration of a Slow-Release Synthetic Prostacyclin Agonist in Rat Fetal Lung Hypoplasia. PLoS One 11:e0161334 (2016). Read more (PubMed: 27529478) »
See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer



The length of time could potentially vary depending on your lab set up. The person who optimized this extraction did not note the exact flow through time so unfortunately I cannot give you an exact recount of how long it took hexane to flow through for us. The flow through rate could depend again on lab set up for the extraction but also the size of column used. The size of column used could also vary depending on the amount of sample you have to run. For a more accurate indication of flow through rates and column size options (most columns have a max volume/ug of protein load) for your sample you will be best served to contact the column manufacturer directly. Below is the link for the column manufacturer we use for C18 columns and we often use cat#9002. They should be able to give you more specific information as well.
http://www51.honeywell.com/sm/rlss/bandj/common/documents/ChromatographyFINAL.pdf

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Answer

Thank you for your enquiry.

The ab133032 - urinary prostacyclin ELISA kit assay is species independent and can be used for any species as long as there is enough prostacyclin present for the kit to detect. Thus, mouse and pig will be fine to use for this assay assuming that the kit will be sensitive enough to detect any expected endogenous levels of this marker in those samples.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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