Description
Human FG beta (Fibrinogen Beta) ELISA Kit
The Human FG Beta (Fibrinogen Beta) ELISA Kit is a powerful tool for measuring levels of fibrinogen beta in human biological samples such as serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring accurate and precise results for a variety of research applications.Fibrinogen beta, a key component of the blood clotting process, plays a crucial role in maintaining hemostasis and preventing excessive bleeding. Abnormal levels of fibrinogen beta have been associated with various health conditions, including cardiovascular diseases, inflammatory disorders, and coagulation disorders.
By using the Human FG Beta ELISA Kit, researchers can delve into the intricate mechanisms of fibrinogen beta regulation and its implications in disease pathogenesis. This kit provides a reliable platform for studying fibrinogen beta as a potential biomarker for disease diagnosis, prognosis, and therapeutic interventions.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection Method: | Colormetric |
| Detection Range: | 1.56-100 ng/mL |
| Sensitivity: | 0.94 ng/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human FG beta in samples. No significant cross-reactivity or interference between Human FG beta and analogues was observed. |
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human FG beta. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human FG beta and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human FG beta, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human FG beta. The concentration of Human FG beta in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | FGB: Fibrinogen has a double Function: yielding monomers that polymerize into fibrin and acting as a cofactor in platelet aggregation. Defects in FGB are a cause of congenital afibrinogenemia (CAFBN). This rare autosomal recessive disorder is characterized by bleeding that varies from mild to severe and by complete absence or extremely low levels of plasma and platelet fibrinogen. Patients with congenital fibrinogen abnormalities can manifest different clinical pictures. Some cases are clinically silent, some show a tendency toward bleeding and some show a predisposition for thrombosis with or without bleeding. |
| UniProt Protein Details: | Protein type:Cell surface; Secreted; Secreted, signal peptide; Adaptor/scaffold Chromosomal Location of Human Ortholog: 4q28 Cellular Component: extracellular space; cell surface; fibrinogen complex; extracellular region; plasma membrane; cell cortex; vesicle; external side of plasma membrane Molecular Function:protein binding, bridging; protein binding; chaperone binding; cell adhesion molecule binding; structural molecule activity; receptor binding Biological Process: protein polymerization; platelet activation; extracellular matrix organization and biogenesis; positive regulation of heterotypic cell-cell adhesion; cell-matrix adhesion; signal transduction; platelet degranulation; cellular protein complex assembly; positive regulation of protein secretion; positive regulation of vasoconstriction; innate immune response; response to calcium ion; blood coagulation; positive regulation of exocytosis Disease: Afibrinogenemia, Congenital; Dysfibrinogenemia, Congenital |
| NCBI Summary: | The protein encoded by this gene is the beta component of fibrinogen, a blood-borne glycoprotein comprised of three pairs of nonidentical polypeptide chains. Following vascular injury, fibrinogen is cleaved by thrombin to form fibrin which is the most abundant component of blood clots. In addition, various cleavage products of fibrinogen and fibrin regulate cell adhesion and spreading, display vasoconstrictor and chemotactic activities, and are mitogens for several cell types. Mutations in this gene lead to several disorders, including afibrinogenemia, dysfibrinogenemia, hypodysfibrinogenemia and thrombotic tendency. Alternatively spliced transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jun 2014] |
| UniProt Code: | P02675 |
| NCBI GenInfo Identifier: | 399492 |
| NCBI Gene ID: | 2244 |
| NCBI Accession: | P02675. 2 |
| UniProt Secondary Accession: | P02675,Q32Q65, Q3KPF2, A0JLR9, B2R7G3, |
| UniProt Related Accession: | P02675 |
| Molecular Weight: | 55,928 Da |
| NCBI Full Name: | Fibrinogen beta chain |
| NCBI Synonym Full Names: | fibrinogen beta chain |
| NCBI Official Symbol: | FGB |
| NCBI Official Synonym Symbols: | HEL-S-78p |
| NCBI Protein Information: | fibrinogen beta chain; fibrinogen, B beta polypeptide; epididymis secretory sperm binding protein Li 78p |
| UniProt Protein Name: | Fibrinogen beta chain |
| Protein Family: | Fibrinogen |
| UniProt Gene Name: | FGB |
| UniProt Entry Name: | FIBB_HUMAN |
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration (ng/mL) | O.D | Average | Corrected |
| 100 | 2.388 2.442 | 2.415 | 2.332 |
| 50 | 1.591 1.647 | 1.619 | 1.536 |
| 25 | 0.921 0.919 | 0.92 | 0.837 |
| 12.5 | 0.482 0.5 | 0.491 | 0.408 |
| 6.25 | 0.249 0.243 | 0.246 | 0.163 |
| 3.13 | 0.186 0.166 | 0.176 | 0.093 |
| 1.56 | 0.122 0.14 | 0.131 | 0.048 |
| 0 | 0.08 0.086 | 0.083 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human FG beta were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human FG beta were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 4.77 | 8.56 | 40.81 | 4.33 | 9.23 | 36.73 |
| Standard deviation | 0.24 | 0.36 | 2.17 | 0.26 | 0.55 | 1.71 |
| C V (%) | 5.03 | 4.21 | 5.32 | 6.00 | 5.96 | 4.66 |
Recovery
The recovery of Human FG beta spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
| Serum (n=5) | 88-98 | 93 |
| EDTA plasma (n=5) | 87-101 | 94 |
| Cell culture media (n=5) | 89-104 | 97 |
Linearity
Samples were spiked with high concentrations of Human FG beta and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
| 1:2 | Range (%) | 95-110 | 92-108 | 90-106 |
| Average (%) | 102 | 99 | 97 | |
| 1:4 | Range (%) | 88-98 | 84-97 | 88-101 |
| Average (%) | 93 | 91 | 94 | |
| 1:8 | Range (%) | 92-104 | 81-93 | 83-96 |
| Average (%) | 97 | 88 | 90 | |
| 1:16 | Range (%) | 92-109 | 83-95 | 87-101 |
| Average (%) | 100 | 88 | 94 |
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
| Micro ELISA Plate(Dismountable) | 8 wells ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent | 1 vial, 10 mL | 4°C(shading light) |
| Stop Solution | 1 vial, 10 mL | 4°C |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.
