| Applications: |
ELISA |
| Reactivity: |
Human |
| Note: |
FOR SCIENTIFIC EDUCATIONAL RESEARCH USE ONLY (RUO). MUST NOT BE USED IN DIAGNOSTIC OR OTHER MEDICAL APPLICATIONS. |
| Sensitivity: |
5.63ng/mL |
| Detection Limit: |
9.38~600ng/mL |
| Short Description: |
This F2 Sandwich ELISA is an in-vitro enzyme-linked immunosorbent assay for the measurement of samples in human cell culture supernatant, serum and plasma (EDTA, citrate, heparin). |
| Storage Instruction: |
If unopened the kit may be stored at 2-8°C for up to 1 month. If the kit will not be used within 1 month, store the components separately, according to the component table in the manual. |
| Assay Time: |
3.5h |
| Detection: |
Colormetric |
| Gene Symbol: |
F2 |
| Gene ID: |
2147 |
| Uniprot ID: |
THRB_HUMAN |
| Specificity: |
This kit recognizes Human PT in samples. No significant cross-reactivity or interference between Human PT and analogues was observed. |
| Sample Type: |
Serum, plasma and other biological fluids |
| Post Translational Modifications | The gamma-carboxyglutamyl residues, which bind calcium ions, result from the carboxylation of glutamyl residues by a microsomal enzyme, the vitamin K-dependent carboxylase. The modified residues are necessary for the calcium-dependent interaction with a negatively charged phospholipid surface, which is essential for the conversion of prothrombin to thrombin. N-glycosylated. N-glycan heterogeneity at Asn-121: Hex3HexNAc3 (minor), Hex4HexNAc3 (minor) and Hex5HexNAc4 (major). At Asn-143: Hex4HexNAc3 (minor) and Hex5HexNAc4 (major). In the penultimate step of the coagulation cascade, prothrombin is converted to thrombin by the prothrombinase complex composed of factor Xa (F10), cofactor Va (F5), and phospholipids. This activation requires factor Xa-catalyzed sequential cleavage at 2 sites, Arg-314 and Arg-363, along 2 possible pathways. In the first pathway, the first cleavage occurs at Arg-314, leading to the formation of the inactive intermediate prethrombin-2. This pathway preferentially occurs on platelets and in the absence of cofactor Va. In the second pathway, the first cleavage occurs at Arg-363, which separates protease domain into 2 chains that remain connected through a disulfide bond and generates the active intermediate meizothrombin. The presence of cofactor Va directs activation along the meizothrombin pathway and greatly accelerates the rate of cleavage at Arg-363, but has a smaller effect on the cleavage of meizothrombin at Arg-314. Meizothrombin accumulates as an intermediate when prothrombinase is assembled on the membrane of red blood cells. |
| Function | Thrombin, which cleaves bonds after Arg and Lys, converts fibrinogen to fibrin and activates factors V, VII, VIII, XIII, and, in complex with thrombomodulin, protein C. Functions in blood homeostasis, inflammation and wound healing. |
| Protein Name | ProthrombinCoagulation Factor Ii Cleaved Into - Activation Peptide Fragment 1 - Activation Peptide Fragment 2 - Thrombin Light Chain - Thrombin Heavy Chain |
| Database Links | Reactome: R-HSA-140837Reactome: R-HSA-140875Reactome: R-HSA-159740Reactome: R-HSA-159763Reactome: R-HSA-159782Reactome: R-HSA-202733Reactome: R-HSA-375276Reactome: R-HSA-381426Reactome: R-HSA-416476Reactome: R-HSA-456926Reactome: R-HSA-76009Reactome: R-HSA-9657688Reactome: R-HSA-9672391Reactome: R-HSA-977606 |
| Cellular Localisation | SecretedExtracellular Space |
| Alternative ELISA Names | Prothrombin ELISA kitCoagulation Factor Ii Cleaved Into - Activation Peptide Fragment 1 - Activation Peptide Fragment 2 - Thrombin Light Chain - Thrombin Heavy Chain ELISA kitF2 ELISA kit |
| Specificity | This kit recognizes Human PT in samples. No significant cross-reactivity or interference between Human PT and analogues was observed. |
| Reproducibility | Both intra-CV and inter-CV are |
Information sourced from Uniprot.org
| Item | Specifications | Storage |
| Micro ELISA Plate (Dismountable) | 96T: 8 wells ×12 strips strips | -20℃, 6 months |
| Reference Standard | 96T: 2 vials 48T: 1 vial | -20℃, 6 months |
| Concentrated Biotinylated Detection Ab (100×) | 96T: 1 vial, 120 μL 60 μL | -20℃, 6 months |
| Concentrated HRP Conjugate (100×) | 96T: 1 vial, 120 μL 60 μL | -20℃ (Protect from light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 2-8°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | 2-8°C, 6 months |
| HRP Conjugate Diluent | 1 vial, 14 mL | 2-8°C, 6 months |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | 2-8°C, 6 months |
| Substrate Reagent | 1 vial, 10 mL | 2-8℃ (Protect from light) |
| Stop Solution | 1 vial, 10 mL | 2-8°C |
| Plate Sealer | 5 pieces | |
| Manual | 1 copy | |
| Certificate of Analysis | 1 copy | |
| Sample Type | Range (%) | Average Recovery (%) |
| Serum(n=8) | 87-103 | 94 |
| EDTA plasma (n=8) | 91-106 | 97 |
| Cell culture media (n=8) | 93-106 | 99 |
| | Intra-assay Precision | Intra-assay Precision | Intra-assay Precision | Inter-assay Precision | Inter-assay Precision | Inter-assay Precision |
| Sample | 1.00 | 2.00 | 3.00 | 1.00 | 2.00 | 3.00 |
| n | 20.00 | 20.00 | 20.00 | 20.00 | 20.00 | 20.00 |
| Mean (ng/mL) | 28.24 | 72.85 | 262.16 | 30.55 | 68.88 | 263.52 |
| Standard deviation | 1.52 | 4.14 | 14.39 | 1.96 | 3.35 | 13.70 |
| CV (%) | 5.38 | 5.68 | 5.49 | 6.42 | 4.86 | 5.20 |
12 months for antibodies. 6 months for ELISA Kits. Please see website T&Cs for further guidance
