Overview

  • Product name

    Factor X Human Chromogenic Activity Assay Kit
  • Detection method

    Colorimetric
  • Sample type

    Cell culture supernatant, Plasma
  • Assay type

    Enzyme activity (quantitative)
  • Sensitivity

    = 0.125 µg/ml
  • Range

    0.125 µg/ml - 8 µg/ml
  • Species reactivity

    Reacts with: Human
  • Product overview

    Factor X Human Chromogenic Activity Assay Kit (ab108833) is developed to determine human Factor X activity in plasma and cell culture. The assay measures the activation of zymogen Factor X to Factor Xa by RVV. The amidolytic activity of the Factor Xa is quantitated using a highly specific Factor Xa substrate releasing a yellow para-nitroaniline (pNA) chromophore. The change in absorbance of the pNA at 405 nm is directly proportional to the Factor X enzymatic activity.


     


    High level of FX activity samples: range of 0.125-8 µg/ml Low level of FX activity samples: range of 0.0156-1 µg/ml


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

  • Notes

    Factor X is a plasma serine protease zymogen involved in the blood coagulation cascade. Factor X is purified from plasma as a two-chain protein consisting of a 45-kDa heavy chain and a 17-kDa light chain. The Factor X heavy chain is cleaved during coagulation by several different proteases including the intrinsic Xase complex, the Factor X-activating enzyme from Russell’s viper venom (RVV) and trypsin, and also by extrinsic (tissue Factor/Factor VIIa) pathway to give an active enzyme Factor Xa. Factor Xa as the activator of prothrombin occupies a central position linking the two blood coagulation pathways.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 100 tests
    Assay Diluent 1 x 5ml
    Factor X Standard 1 unit
    Factor Xa Substrate 2 vials
    Microplate 1 unit
    RVV 1 vial
    Sample Diluent 1 x 5ml
    Sealing Tapes 3 units
  • Research areas

  • Function

    Factor Xa is a vitamin K-dependent glycoprotein that converts prothrombin to thrombin in the presence of factor Va, calcium and phospholipid during blood clotting.
  • Tissue specificity

    Plasma; synthesized in the liver.
  • Involvement in disease

    Factor X deficiency
  • Sequence similarities

    Belongs to the peptidase S1 family.
    Contains 2 EGF-like domains.
    Contains 1 Gla (gamma-carboxy-glutamate) domain.
    Contains 1 peptidase S1 domain.
  • Post-translational
    modifications

    The vitamin K-dependent, enzymatic carboxylation of some glutamate residues allows the modified protein to bind calcium.
    N- and O-glycosylated. O-glycosylated with core 1 or possibly core 8 glycans.
    The activation peptide is cleaved by factor IXa (in the intrinsic pathway), or by factor VIIa (in the extrinsic pathway).
    The iron and 2-oxoglutarate dependent 3-hydroxylation of aspartate and asparagine is (R) stereospecific within EGF domains.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • Activated factor Xa heavy chain
    • F10
    • FA10_HUMAN
    • Stuart factor
    • Stuart-Prower factor
    see all
  • Database links

Images

  • Factor X Chromogenic Activity measured in cell culture supernatants showing quantity (ug) per mL of tested sample after 40 min of incubation. Values calculated based on Low Level Standard Curve. Samples were diluted 2-10 fold.

  • Factor X Chromogenic Activity measured in biological fluids showing quantity (ug) per mL of tested sample after 10 min of incubation. Values calculated based on High Level Standard Curve. Samples were diluted 2 fold.

  • Standard curve: mean of duplicates (+/- SD) with background reads subtracted

  • Standard curve: mean of duplicates (+/- SD) with background reads subtracted

Protocols

References

ab108833 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Thank you for your call last week and for your patience. I apologize for the extended delay in getting this information for you.

I've just received a reply from the lab, and these kits can be analyzed at room temperature. The OD might be slightly lower than at 37C but the kinetics should not be affected. If you warm your samples to 37C prior to analysis and then read the plate at room temperature, there should not be a noticeable effect. We would still recommend analyzing the kinetic data as opposed to an end point analysis.

I hope this information will be useful, but please let me know if you have any further questions or if there is anything else that we can do for you. I'll be happy to help.

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Question
Answer

Thank you for your call today.

As we discussed, you can email mailto:us.sales@abcam.com for information regarding bulk purchases.

Please let me know if you have any questions or if there is anything else that we can do for you.

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Answer

Thank you for your reply.

Unfortunately we are unable to run inhibitor tests for our customers.

However, if I can help you in any other way please do letme know.

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Answer

Thank you for contacting us and your interest in our products. I am sorry for the delay in getting back to you.


The Factor VII Human Chromogenic Activity Assay Kit had been developed to measure human FVII activity in plasma and cell culture supernatants. The assay works by binding the FVII present in the sample to the plate by a monoclonal antibody pre-coated on the plate. The activity of the FVII is then quantified by introducing a FXa substrate which upon cleavage releases ayellow para-nitroaniline (pNA) chromophore. The change in absorbance of the pNA at 405 nm is directly proportional to the FVII enzymatic activity.


The Factor X Human Chromogenic Activity Assay Kit (ab108833) similarlymeasures the activation of zymogen Factor X to Factor Xa by RVV. The amidolytic activity of the Factor Xa isquantified using a highly specific Factor Xa substrate releasing a yellow para-nitroaniline (pNA) chromophore. The change in absorbance of the pNA at 405 nm is directly proportional to the Factor X enzymatic activity.


We have had reports of customers using these kits to test inhibitors. We have not performed this ourselves in-house and some optimisationmay therefore be required. It may be that as you want to measure the inhibition of activity of your compounds, not the Factor VII or X activity within a biological sample, for which the kit is designed, you may need more Factor X and VII standard than what the kit provides.


The standard provided with the kit is enough for two sets of duplicate standard curve. If you require more we may be able to sell the FVII and Factor X standard separately.If you would like me to look into this please let me know.


I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

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Question
Answer

Thank you for your recent telephone enquiry. I am sorry to confirm that this kit has not been tested in mouse. Due to the nature of the kit, it is difficult to predict whether it would still work on mouse samples. I am sorry we do not have more information for you on this occasion. If you have any further questions, please do not hesitate to contact us. 

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