|Size||96wells/kit, with removable strips.|
|Sample Type||cell culture supernates, serum, plasma(heparin, EDTA, citrate) and urine.|
|Product Name||Human Thrombomodulin PicoKine™ ELISA Kit|
|Storage & Handling||Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)|
|Size||96wells/kit, with removable strips.|
|Description||Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human Thrombomodulin. 96wells/kit, with removable strips.|
|Cite This Product||Human Thrombomodulin PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0917)|
|Sample Type||cell culture supernates, serum, plasma(heparin, EDTA, citrate) and urine..
Anticoagulant(s): heparin, EDTA or Citrate
*The recommended anticoagulants are proven to not block the antibody binding sites on the target antigen. Please do not collect blood sample with other anticoagulants that are not specified above or contact us to check for feasibility.
|Immunogen||Expression system for standard: NSO; Immunogen sequence: A19-S515|
|Cross Reactivity||There is no detectable cross-reactivity with other relevant proteins.|
|Antibody Clonalities||Capture Antibody | Detection Antibody:
monoclonal antibody from mouse|polyclonal antibody from goat
|EK0917-CAP||96-well plate precoated with anti-Human THBD antibody||1|
|EK0917-ST||lyophilized recombinant Human THBD standard||10ng/tube|
|EK0917-DA||biotinylated anti-Human THBD antibody||130ul|
|AR1106-1||sample diluent buffer||30ml|
|AR1106-2||antibody diluent buffer||12ml|
|AR1106-3||ABC diluent buffer||12ml|
|AR1104||TMB color developing agent||10ml|
|AR1105||TMB stop solution||10ml|
*Why there is no wash buffer? Our Avidin-Biotin-Peroxidase Diluent contains the detergent (TWEEN) normally present in other companies' ELISA kits. This saves you the step of having to wash with the special wash buffer and achieve similar or better signal to noise ratio. The wash can use regular wash buffers (PBS, TBS etc.) commonly found in labs.
Materials Required But Not Provided
- 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; 450μl of purified acetic acid or 700μl of concentrated hydrochloric acid to 1000ml H2
- Preparation of 0.01 M PBS: Add 8.5g sodium chloride, 1.4g Na O and adjust pH to 7.2-7.
Typical Data Obtained from Human Thrombomodulin PicoKine™ ELISA Kit
(TMB reaction incubate at 37°C for 15-25min)
Intra/Inter Assay Precision
|Intra-Assay Precision||Inter-Assay Precision|
Three samples with differing target protein concentrations were assayed using four different lots to measure the CV% lot to lot variance.
To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots.
|Lots||Lot1 (pg/ml)||Lot2 (pg/ml)||Lot3 (pg/ml)||Lot4 (pg/ml)||Mean (pg/ml)||Standard Deviation||CV (%)|
*The typical data is obtained from Boster's internal QC result and for reference only. It may differ from the lab test results of the end users. It is more important that the user's lab test results reflect the same linearity demonstrated in the typical data than achieving exactly the same O.D. values.
Protein Target Info (Source: Uniprot.org)
|Tissue Specificity||Endothelial cells are unique in synthesizing thrombomodulin.|
|Subcellular Localization||Membrane; Single-pass type I membrane protein.|
|Molecular Weight||60329 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|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.|
|Research Areas||Cardiovascular, Stem Cells
*You can search these to find other products in these research areas.
|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 74kDa, 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).|
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1. Diluent the samples with the provided sample diluent buffer into 100ul.
2. Add 50ul of standard solution, when 50ul of sample will be added into a well.
• Add tissue homogenates into the wells and then add ABC and TMB without adding any biotinylated detection antibody to see if any signal will be observed.
• If no signal is produced, then you can work on the tissue sample by using the kit.
The 1XTBS can be used if the pH value falls in the range of 7.2-7.6.
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