Overview

  • Product name

    Human Antithrombin III ELISA Kit
    See all Antithrombin III kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 4.8%
    Inter-assay
    Sample n Mean SD CV%
    Overall 7.1%
  • Sample type

    Cell culture supernatant, Saliva, Milk, Urine, Serum, Plasma, Cell culture media, Cerebral Spinal Fluid
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    = 3 ng/ml
  • Range

    3.13 ng/ml - 200 ng/ml
  • Recovery

    98.5 %

  • Assay time

    4h 0m
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Abcam’s Antithrombin III Human in vitro ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Antithrombin III in urine, plasma, milk, saliva, cerebrospinal fluid, serum, and cell culture supernatants.


    An antithrombin III specific antibody has been precoated onto 96-well plates and blocked. Standards or test samples are added to the wells and subsequently an antithrombin III specific biotinylated detection antibody is added and then followed by washing with wash buffer. Streptavidin-Peroxidase Complex 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 antithrombin III 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 ab108801 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

  • Linearity: Antithrombin III in samples at different diutions, corrected for the factor of dilution  (duplicates +/- SD)

  • Linearity: Antithrombin III in samples at different diutions, corrected for the factor of dilution (duplicates +/- SD), showing ng x mL-1 on y-axis.

  • Linearity: Antithrombin III in samples at different diutions, corrected for the factor of dilution  (duplicates +/- SD)

  • Antithrombin III in cell culture supernatants (duplicates +/- SD), tested in the dilution range of 1:1-1:100.

  • Antithrombin III in human biological fluids (duplicates +/- SD). The samples were differently diluted  (serum and Li-heparine plasma: 1e4-1e5; CSF: 1e2-1e3; milk: 10-100; saliva: 5-100; urine: neat)

  • Representative standard curve using ab108801.

Protocols

References

ab108801 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

Please find below the information on how to prepare the standard for lots GR116626-1, Lot GR115289-1 and Lot GR106377-1 of the Antithrombin III Human ELISA Kit ab108801:


· MIX Diluent Concentrate (10x): If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved. Dilute the MIX Diluent Concentrate 1:10 with reagent grade water. Store for up to 1 month at 2 - 8ºC.

· AT III Standard: Reconstitute the 4.8 ug of human AT III Standard with 4 ml of MIX Diluent to generate a stock solution of 1.2 ug/ml. Allow the standard to sit for 10 minutes with gentle agitation prior to making dilutions. The stock solution (1.2 ug/ml) should be further diluted 1:6 with MIX Diluent to produce 200 ng/ml standard solution. Prepare duplicate or triplicate standard points by serially diluting the Standard solution (200 ng/ml) 1:2 with equal volume of MIX Diluent to produce 100, 50, 25, 12.5, 6.25, and 3.13 ng/ml. MIX Diluent serves as the zero standard (0 ng/ml). Any remaining solution should be frozen at -20ºC and used within the next 30 days.






Standard Point
Dilution
[AT III] (ng/ml)

P1
1 part stock (1.2 ug/ml) + 5 parts MIX Diluent
200.00

P2
1 part P1 + 1 part MIX Diluent
100.00

P3
1 part P2 + 1 part MIX Diluent
50.00

P4
1 part P3 + 1 part MIX Diluent
25.00

P5
1 part P4 + 1 part MIX Diluent
12.50

P6
1 part P5 + 1 part MIX Diluent
6.25

P7
1 part P6 + 1 part MIX Diluent
3.13

P8
MIX Diluent
0.00









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Answer

Thank for your email. I am sorry to hear that you have had bad experience with Abcam. Generally we are very flexible with our customers - our customer comes first than business so I am sure this time you will have the best technical support experience.

The kit ab108801 is recommended to be used with 96 well plate. The experiment is set up in such a way that you would need 12 wells (6 in duplicate) for standard curve or 14 as given in table on page 9 and 2 wells for blank (duplicate). Now you are left with 80 wells.

The 80 wells in one plate can be used for 40 samples in duplicate which is recommended or 80 samples if you are comfortable with pipetting etc.

426 - 40 (in duplicate with one kit) = 386 samples

If you are interested in duplicate then 40 samples are possible with first kit and 48 samples with each rest of the kits (96 wells to be used with each kit) in total 8. This means you would need 9 kits in total.

My recommendation is to try the standard curve in at least one other kit as well to see the sample variations. In this case you would need total of 10 kits.

Could you let me know if you are interested in buying 9-10 kits as recommended or have different opinion. I will then be happy to quote the kits with discount values.

Thanks!

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