Overview

  • Product name

    Tissue Factor Activity Assay Kit (Human, Colorimetric)
    See all Tissue Factor kits
  • Detection method

    Colorimetric
  • Sample type

    Urine, Serum, Plasma, Cell culture extracts, Tissue
  • Assay type

    Enzyme activity (quantitative)
  • Assay time

    1h 00m
  • Species reactivity

    Reacts with: Human
  • Product overview

    Tissue Factor Activity Assay Kit (Human, Colorimetric) ab108906 is used to determine Human Tissue Factor activity in plasma, tissue and cell culture supernatants.


    The tissue factor activity assay protocol measures the ability of lipoprotein TF/FVIIa to activate factor X (FX) to factor Xa. The amidolytic activity of the TF/FVIIa complex is quantitated by the amount of FXa produced using a highly specific FXa substrate releasing a yellow para-nitroaniline (pNA) chromophore. The change in absorbance of the pNA at 405 nm is directly proportional to the TF enzymatic activity.


    Tissue factor activity assay protocol summary:
    - add reaction mix to wells
    - add standards and samples
    - incubate for 30 min
    - add FXa substrate
    - analyze with a microplate reader every 5 min for 25 min


    The entire kit may be stored at -20°C for long term storage before reconstitution - Avoid repeated freeze-thaw cycles.

  • Notes

    Previously called Tissue Factor Human Chromogenic Activity Assay Kit.

    The transmembrane protein Tissue factor (TF) is the physiologic trigger of coagulation in normal hemostasis. Tissue factor binds and allosterically activates factor VII (FVII). The TF-FVIIa complex cleaves factor IX and X, leading to thrombin generation. Tissue factor markedly enhances the ability of FVIIa to cleave both macromolecule and small peptidyl substrates.

  • Platform

    Microplate reader

Properties

Images

  • Colorimetric standard curve: mean of duplicates (+/- SD)

  • Standard curve showing concentration (pM) on X-axis and increase of O.D. per min on Y-axis (time frame: 20 min.)

  • Time course showing O.D.'s of one standard point and one sample (undiluted vs.diluted)

  • Tissue Factor measured in various biologicals showing average concentrations

Protocols

References

This product has been referenced in:

  • Schulz A  et al. Protective vascular coagulation in response to bacterial infection of the kidney is regulated by bacterial lipid A and host CD147. Pathog Dis N/A:N/A (2018). Read more (PubMed: 30476069) »
  • Watanabe Y  et al. Indoleamine 2,3-dioxygenase 1 in coronary atherosclerotic plaque enhances tissue factor expression in activated macrophages. Res Pract Thromb Haemost 2:726-735 (2018). Read more (PubMed: 30349892) »
See all 4 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Question
Answer


The standard should be reconstituted in 3ml water.

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Answer



After arrival please store the individual components as instructed in the protocol booklet:

ab108906 may be stored until the expiration date at 2 to 8°C from the date of shipment. Store Standard, Factor VII protein, Factor X protein and FXa Substrate at -200C. Store Microplate, Sample Diluent, Assay Diluent at 2-80C. Opened Diluent may be stored for up to 1 month at 2 to 8°C.

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Answer

Since tissue factor is both secreted and membrane bound, we recommend using cell lysates so the protein is properly released from the membrane.

As mentioned in the protocol under step 6.2) Cells lysates can be prepared as follows:

1. Lyse and solubilize cultured cells with 15 mM octyl-β-D-glucopyranoside at 37°C for 15 minutes.
2. Collect fresh cell lysates and assay.
3. Samples can be stored at -20°C or below for 3 months.

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Question
Answer


The labs says that the kit contains a low binding plate which will be better. However, otherwise the brand of the 96-well plate does not matter.

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Answer

Thank you for contacting us on Friday in regards to the Tissue Factor Human Chromogenic Activity Assay Kit (ab108906).

I can now inform you that the vial of Tissue Factor Standard you received required the addition of 1.2 ml of reagent grade water to generate a solution of 250 pM.

We would advise allowing the standard to sit for 10 minutes with gentle agitation prior to making dilutions.

I hope this information has been of help. I thank you for bringing this problem to our attention, we will be updating the protocol booklet to reflect this.

If you have any further questions, please do not hesitate to ask.

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Answer


The amount of standard is only provided on the vial label. The datasheet and the protocol bookletare only generic.
Please ask your customer tocheck the vial label, from which they can work out the volume of buffer they need to to generate a standard solution of 250 pM.

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Answer

Thank you very much for contacting us.
I just wanted to let you know that I am still working on your inquiry. I am looking forthe information you requested.
I will forward to you the information as soon as I receive it. Thank you for your patience.

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Answer


I have just been informed that the tissue factor standard should be reconstituted with 1.2 ml of reagent grade water to generate a solution of 250 pM.

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