Overview

  • Product name

    Mouse TFPI ELISA Kit
    See all TFPI kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Mouse serum 8 3.7%
    Inter-assay
    Sample n Mean SD CV%
    Mouse serum 3 7.2%
  • Sample type

    Cell culture supernatant, Urine, Serum, Tissue Extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    1.7 pg/ml
  • Range

    18.75 pg/ml - 1200 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 99 97% - 102%
    Serum 102 96% - 109%
    Tissue Extracts 92 85% - 101%
    Cell culture media 98 96% - 101%
    Heparin Plasma 109 104% - 112%
    EDTA Plasma 101 96% - 105%
    Citrate Plasma 103 98% - 111%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Rat, Cow, Human
  • Product overview

    TFPI in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of mouse TFPI protein in serum, plasma, urine, cell culture supernatant, and tissue extract samples.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Sensitivity:


    Samples in 1X Cell Extraction Buffer PTR - 1.7 pg/mL


    Samples in Sample Diluent NS - 1.87 pg/mL


     

  • Notes

    Mouse TFPI (tissue factor pathway inhibitor) is a secreted protein located on the endothelial surface, within platelets, in plasma, and on monocytes. Functionally, mouse TFPI is the primary inhibitor of intravascular tissue factor (TF) activity. TF is the principal protein that initiates blood coagulation in vivo. Thus, Mouse TFPI inhibits coagulation.

    Mouse TFPI has an acidic N-terminal region, three Kunitz (K) domains separated by two linker regions, and a C-terminal basic region. The first K domain inhibits TF from binding coagulation factor VIIa. The second K domain inhibits factor Xa. The third K domain binds to heparin. Human and rat TFPI exhibit 58% and 71% amino acid identity to mouse TFPI, respectively.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    Mouse TFPI Lyophilized Recombinant Protein 2 vials
    10X Mouse TFPI Detector Antibody 1 x 600µl
    10X Mouse TFPI Capture Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPR 1 x 6ml
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Inhibits factor X (X(a)) directly and, in a Xa-dependent way, inhibits VIIa/tissue factor activity, presumably by forming a quaternary Xa/LACI/VIIa/TF complex. It possesses an antithrombotic action and also the ability to associate with lipoproteins in plasma.
  • Tissue specificity

    Mostly in endothelial cells.
  • Sequence similarities

    Contains 3 BPTI/Kunitz inhibitor domains.
  • Domain

    This inhibitor contains three inhibitory domains. The first domain interacts with VIIa and TF, the second one with Xa.
  • Post-translational
    modifications

    O-glycosylated.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • Anti convertin
    • EPI
    • Extrinsic pathway inhibitor
    • LACI
    • Lipoprotein associated coagulation inhibitor
    • Lipoprotein-associated coagulation inhibitor
    • TFI
    • TFPI
    • TFPI 1
    • TFPI1
    • TFPI1_HUMAN
    • Tissue factor pathway inhibitor
    • Tissue factor pathway inhibitor (lipoprotein associated coagulation inhibitor)
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab217776 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of TFPI were measured in duplicates, interpolated from the TFPI standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:100, plasma (citrate) 1:400, plasma (EDTA) 1:400 and plasma (heparin) 1:400. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TFPI concentration was determined to be 57.10 ng/mL in neat serum, 241.24 ng/mL in neat plasma (citrate), 259.59 ng/mL in neat plasma (EDTA) and 303.08 ng/mL in neat plasma (heparin).

  • The concentrations of TFPI were measured in duplicates, interpolated from the TFPI standard curves and corrected for sample dilution. Undiluted samples are as follows: urine 50%, and cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TFPI concentration was determined to be 120.64 pg/mL in native, neat urine, 1,353.96 pg/mL in spiked, neat urine and 1,130.58 pg/mL in spiked, neat cell culture media.

  • The concentrations of TFPI were measured in duplicate and interpolated from the TFPI standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TFPI concentration was determined to be 525.44 pg/mL in mouse heart extract and 69.55 pg/mL in mouse liver extract.

Protocols

References

ab217776 has not yet been referenced specifically in any publications.

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