Overview

  • Product name

    Human Thrombin ELISA Kit (Factor II)
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 4.7%
    Inter-assay
    Sample n Mean SD CV%
    Overall 7.2%
  • Sample type

    Cell culture supernatant
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    = 0.3 ng/ml
  • Range

    0.313 ng/ml - 20 ng/ml
  • Recovery

    98 %

  • Assay time

    4h 0m
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Abcam’s Thrombin (Factor II) Human in vitro ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Thrombin concentrations in cell culture supernatants. This assay recognizes both natural and recombinant Human thrombin.


    A Thrombin specific antibody has been precoated onto 96-well plates and blocked. Standards or test samples are added to the wells and subsequently a Thrombin specific biotinylated detection antibody is added and then followed by washing with wash buffer. Streptavidin-Peroxidase Conjugate is added and unbound conjugates are washed away with wash buffer. TMB is then used to visualize Streptavidin-Peroxidase enzymatic reaction. TMB is catalyzed by Streptavidin-Peroxidase to produce a blue color product that changes into yellow after adding acidic stop solution. The density of yellow coloration is directly proportional to the amount of Thrombin captured in plate.


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

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

Applications

Our Abpromise guarantee covers the use of ab108909 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 dilution.

Images

  • Representative Standard Curve Using ab108909.

Protocols

References

This product has been referenced in:

  • Wang H  et al. N-glycosylation in the protease domain of trypsin-like serine proteases mediates calnexin-assisted protein folding. Elife 7:N/A (2018). Read more (PubMed: 29889025) »
  • Moñux G  et al. FXa inhibition by rivaroxaban modifies mechanisms associated with the pathogenesis of human abdominal aortic aneurysms. Br J Clin Pharmacol 83:2661-2670 (2017). Read more (PubMed: 28735510) »
See all 3 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

The thrombin ELISA, using the Abcam human ELISA kit Ab108909, lot GR232366-8, was performed by an experienced laboratory technician at an accredited university. The goal of the experiment was to analyze the linearity of dilutions of rhesus macaque EDTA-prepared plasma samples and serum samples.

Overall, the kit, like other Abcam ELISA kits, was easy to follow, clear in terms of directions, and brief yet thorough. The laboratory technician had no problems understanding any of the protocol’s steps.

Our test was performed in 2 basic steps. The first was to find the appropriate testable dilution in plasma and serum, and the second was to test the linearity and spike recovery.

To this end our first experiment tested 8 dilutions of both serum and plasma ranging from 1:10 to 1:163840 (8 1:4 serial dilutions). The plasma was not in range of the assay, confirming that his kit does not detect pro-thrombin. The serum dilutions gave OD values in range of the assay from 1:10 – 1:2560, but a linear range was not observed.

We saved the thrombin standard vial at -80C for 1 week between tests.

Our second experiment consisted of a narrower dilution range at 1:50, 1:100, 1:200, 1:400,a nd 1:800 for n=2 rhesus sera and a spike recovery test. The tested range was well within the OD limits of the assay. N=1 rhesus serum may have been near linear range at 1:400-1:800, but n=1 rhesus serum was not. The spike recovery (performed with Serum from animal 2) showed an increasing spike recovery with dilution (40% at 1:50, 80% at 1:800).

Overall, we would recommend this kit for its ease of use. We believe there is positive cross-reactivity for rhesus, but we were unable to identify a linear range for the assay.

Abcam user community

Verified customer

Submitted Jun 07 2016

Answer

Thank you for your patience while I looked into this.

The lab has sent us thereplacement vials of the diluent and wash buffer. We are sending these to you free of charge on order. You should receive these tomorrow.

I hope this information helps. Please contact us with any other questions.

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Question
Answer

Thank you for contacting Abcam regarding ab108909.


Just to confirm what we discussed over the phone:


Thrombin Standard: Purified human Thrombin in a buffered protein base (40 ng, lyophilized).

Standard Curve: Reconstitute the 40 ng of human Thrombin Standard with 2 ml of EIA Diluent to generate 20 ng/ml of stock solution. Allow the standard to sit for 10 minutes with gentle agitation prior to making dilutions. Prepare duplicate or triplicate standard points by serially diluting the Thrombin standard stock (20 ng/ml) 1:2 with equal volume of EIA Diluent to produce 10, 5, 2.5, 1.25, 0.625 and 0.313 ng/ml. EIA Diluent serves as the zero standard (0 ng/ml). Any remaining solution should be frozen at -200C and used within 30 days.


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions or concerns.

Read More

Answer

Thank you for contacting us.

I heard back from the lab and they recommend that youdispose the Wash Buffer and the Diluent if you mixed them together.

Wash Buffer is 20x and should be diluted with water (1 part Wash Buffer + 19 parts of water).
Diluent is 10x and should be diluted with water (1 part of Diluent + 9 parts of water).
Wewill send you 1bottle of the Wash Buffer and 1 bottle of the Diluent free of charge, but it may take us until next week to get these to you.
I will be in touch again shorthly.

Read More

Answer

Thank you for calling Abcam earlier today.
I have talked to the lab and they do not think there is protein the same size as thrombin in the buffer based, but we cannot be 100% sure. The exact components of the buffer are proprietary and so I am unable to provide that information.
Please let me know if there is anything else I can help you with.

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