Thrombomodulin ELISA Kit

Catalog No. ABIN1706068

Human Thrombomodulin ELISA Kit Colorimetric assay for quantification of Human Thrombomodulin and has been mentioned in 1 publication.

Quick Overview for Thrombomodulin ELISA Kit (ABIN1706068)

Target: Thrombomodulin (THBD)

Binding Specificity: AA 19-515

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 62.5-4000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA), Plasma (citrate), Urine

Product Details Thrombomodulin ELISA Kit

Minimum Detection Limit: 62.5 pg/mL

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human Thrombomodulin

Brand: PicoKine™

Analytical Method: Quantitative

Specificity: Expression system for standard: NSO
Immunogen sequence: A19-S515

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <10pg/mL

Material not included: Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl

Immunogen: Expression system for standard: NSO
Immunogen sequence: A19-S515

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Comment:

Sequence similarities: Contains 1 C-type lectin domain.

Tissue Specificity: Endothelial cells are unique in synthesizing thrombomodulin.

Plate: Pre-coated

Protocol: human Thrombomodulin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Thrombomodulin has been precoated onto 96-well plates. Standards(NSO, A19-S515) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Thrombomodulin is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human Thrombomodulin amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 4000pg/mL, 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL human Thrombomodulin standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of human cell culture supernates, serum, plasma(heparin, EDTA, citrate) or urine to each empty well. See "Sample Dilution Guideline" above for details. We recommend that each human Thrombomodulin standard solution and each sample is measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 523, Standard deviation: 24.1, CV(%): 4.6
• Sample 2: n=16, Mean(pg/ml): 1074, Standard deviation: 40.1, CV(%): 3.7
• Sample 3: n=16, Mean(pg/ml): 2246, Standard deviation: 92.1, CV(%): 4.1,
• Sample 1: n=24, Mean(pg/ml): 576, Standard deviation: 47.2, CV(%): 8.2
• Sample 2: n=24, Mean(pg/ml): 1187, Standard deviation: 86.7, CV(%): 7.3
• Sample 3: n=24, Mean(pg/ml): 2315, Standard deviation: 180.6, CV(%): 7.8

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

References for Thrombomodulin ELISA Kit (ABIN1706068)

Kupchak, Kazman, Vingren, Levitt, Lee, Williamson, Armstrong, Deuster: "Blood Hemostatic Changes During an Ultraendurance Road Cycling Event in a Hot Environment." in: Wilderness & environmental medicine, Vol. 28, Issue 3, pp. 197-206, (2018) (PubMed).

Target Details for Thrombomodulin

Target: Thrombomodulin (THBD)

Alternative Name: THBD

Background:

Protein 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.

Background: Thrombomodulin(TM), CD141 or BDCA-3 is an integral membrane protein expressed on the surface of endothelial cells and serves as a cofactor for thrombin. It reduces blood coagulation by converting thrombin to an anticoagulant enzyme from a procoagulant enzyme. Thrombomodulin is also expressed on human mesothelial cell, monocyte and a dendritic cell subset. In humans, thrombomodulin is encoded by the THBD gene. The protein has a molecular mass of 74 kDa, and consists of a single chain with 5 distinct domains. Thrombomodulin functions as a cofactor in the thrombin-induced activation of protein C in the anticoagulant pathway by forming a 1:1 stoichiometric complex with thrombin. This raises the speed of protein C activation thousandfold. Thrombomodulin-bound thrombin has procoagulant effect at the same time by inhibiting fibrinolysis by cleaving thrombin-activatable fibrinolysis inhibitor(TAFI,aka carboxypeptidase B2) into its active form. Thrombomodulin is a glycoprotein on the surface of endothelial cells that, in addition to binding thrombin, regulates C3b inactivation by factor I. Mutations in the thrombomodulin gene(THBD) have also been reported to be associated with atypical hemolytic-uremic syndrome(aHUS).

Synonyms: Thrombomodulin,TM,Fetomodulin,CD141,THBD,THRM,

Full Gene Name: Thrombomodulin

Cellular Localisation: Membrane, Single-pass type I membrane protein.

Gene ID: 7056

UniProt: P07204