Human Thrombomodulin ELISA Kit (ab214029)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 3 pg/ml
- Range: 20.3 pg/ml - 1300 pg/ml
- Sample type: Cell culture extracts, Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum, Tissue Extracts, Urine
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human Thrombomodulin ELISA Kit
See all Thrombomodulin kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Human serum 8 3.1% Inter-assay Sample n Mean SD CV% Human serum 3 9.4% -
Sample type
Cell culture supernatant, Urine, Serum, Cell culture extracts, Tissue Extracts, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
3 pg/ml -
Range
20.3 pg/ml - 1300 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture supernatant 90 88% - 91% Urine 106 102% - 112% Serum 97 96% - 99% Cell culture extracts 94 92% - 96% Tissue Extracts 105 101% - 110% Hep Plasma 99 98% - 100% EDTA Plasma 96 94% - 98% Cit plasma 96 92% - 98% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human
Does not react with: Cow -
Product overview
Human Thrombomodulin ELISA Kit (ab214029) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Thrombomodulin protein in cell culture extracts, cell culture supernatant, cit plasma, edta plasma, hep plasma, serum, urine, and tissue extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Thrombomodulin with 3 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Notes
Human Thrombomodulin is a 557 amino acid type I transmembrane protein, consisting of six epidermal growth factor (EGF)-like domains and is highly glycosylated. Thrombomodulin is an important component in the anti-coagulation system by functioning as a cell surface receptor for thrombin, and has also been shown to play a role in anti-inflammation, cell adhesion, and proliferation. The Thrombomodulin - thrombin complex activates protein C and thrombin-activatable fibrinolysis inhibitor (TAFI); protein C degrades coagulation factors Va and VIIIa, which regulates thrombin resulting in inhibition of coagulation. The extracellular portion of Thrombomodulin has been shown to be released from the cell surface due to protein cleavage during vascular endothelial cell injury.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Wash Buffer PT (ab206977) 1 x 20ml 20X Human Thrombomodulin Detector Antibody 1 x 300µl 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 4BI 1 x 6ml Human Thrombomodulin Capture Antibody (Lyophilized) 1 vial Human Thrombomodulin Lyophilized Recombinant Protein (ab98989) 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 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
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Function
Thrombomodulin is a specific endothelial cell receptor that forms a 1:1 stoichiometric complex with thrombin. This complex is responsible for the conversion of protein C to the activated protein C (protein Ca). Once evolved, protein Ca scissions the activated cofactors of the coagulation mechanism, factor Va and factor VIIIa, and thereby reduces the amount of thrombin generated. -
Tissue specificity
Endothelial cells are unique in synthesizing thrombomodulin. -
Involvement in disease
Defects in THBD are the cause of thrombophilia due to thrombomodulin defect (THR-THBD) [MIM:188040]. A hemostatic disorder characterized by a tendency to thrombosis.
Defects in THBD are a cause of susceptibility to hemolytic uremic syndrome atypical type 6 (AHUS6) [MIM:612926]. An atypical form of hemolytic uremic syndrome. It is a complex genetic disease characterized by microangiopathic hemolytic anemia, thrombocytopenia, renal failure and absence of episodes of enterocolitis and diarrhea. In contrast to typical hemolytic uremic syndrome, atypical forms have a poorer prognosis, with higher death rates and frequent progression to end-stage renal disease. Note=Susceptibility to the development of atypical hemolytic uremic syndrome can be conferred by mutations in various components of or regulatory factors in the complement cascade system. Other genes may play a role in modifying the phenotype. -
Sequence similarities
Contains 1 C-type lectin domain.
Contains 6 EGF-like domains. -
Post-translational
modificationsN-glycosylated.
The iron and 2-oxoglutarate dependent 3-hydroxylation of aspartate and asparagine is (R) stereospecific within EGF domains. -
Cellular localization
Membrane. - Information by UniProt
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Alternative names
- AHUS 6
- AHUS6
- BDCA 3
see all -
Database links
- Entrez Gene: 7056 Human
- Omim: 188040 Human
- SwissProt: P07204 Human
- Unigene: 2030 Human
Images
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Other - Human Thrombomodulin ELISA Kit (ab214029)
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.
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Human Thrombomodulin standard curve comparison data.Standard curve comparison between human Thrombomodulin SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows a 8-fold increase in sensitivity.
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Example of Human Thrombomodulin standard curve in Sample Diluent NS.Background-subtracted data values (mean +/- SD) are graphed.
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Example of Human Thrombomodulin standard curve in 1X Cell Extraction Buffer PTR.Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native Thrombomodulin in human serum, and plasma samples.The concentrations of Thrombomodulin were measured in duplicate, interpolated from the Thrombomodulin standard curve and corrected for sample dilution. Undiluted samples are as follows: serum 25%, plasma (citrate) 25%, plasma (EDTA) 25% and plasma (heparin) 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Thrombomodulin concentration was determined to be 4.8 ng/mL in serum, 3.9 ng/mL in plasma (citrate), 3.8 ng/mL in plasma (EDTA) and 4.3 ng/mL in plasma (heparin).
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Interpolated concentrations of spiked recombinant Thrombomodulin in HepG2 cell culture supernatant samples.The concentrations of Thrombomodulin were measured in duplicate, interpolated from the Thrombomodulin standard curve and corrected for sample dilution. Undiluted sample is at 45%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The native Thrombomodulin concentration was undetectable in the null spike control run in parallel with the spiked sample.
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Interpolated concentrations of native Thrombomodulin in HUVEC and human liver homogenate extracts based on a 1,000 µg/mL extract load.The concentrations of Thrombomodulin were measured in duplicate and interpolated from the Thrombomodulin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Thrombomodulin concentration was determined to be 1.2 ng/mL in HUVEC extract and 8.0 ng/mL in liver homogenate extract.
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Serum from ten individual healthy human male donors was measured in duplicate.Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Thrombomodulin concentration was determined to be 5.1 ng/mL with a range of 3.6 – 6.8 ng/mL.
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Serum from ten individual healthy human female donors was measured in duplicate.Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean target concentration was determined to be 5.1 ng/mL with a range of 2.7 – 9.9 ng/mL.
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Sandwich ELISA - Human Thrombomodulin ELISA Kit (ab214029)To learn more about the advantages of recombinant antibodies see here.
