Human Fibrinogen ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human Fibrinogen with a sensitivity of 29 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
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Key facts
- Detection method
- Fluorescent
- Sample types
- Saliva, Urine, Heparin Plasma, Citrate plasma, Cell culture supernatant, Milk, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 15.63 - 64000 pg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 29 pg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Target data
Function
Cleaved by the protease thrombin to yield monomers which, together with fibrinogen beta (FGB) and fibrinogen gamma (FGG), polymerize to form an insoluble fibrin matrix. Fibrin has a major function in hemostasis as one of the primary components of blood clots. In addition, functions during the early stages of wound repair to stabilize the lesion and guide cell migration during re-epithelialization. Was originally thought to be essential for platelet aggregation, based on in vitro studies using anticoagulated blood. However, subsequent studies have shown that it is not absolutely required for thrombus formation in vivo. Enhances expression of SELP in activated platelets via an ITGB3-dependent pathway. Maternal fibrinogen is essential for successful pregnancy. Fibrin deposition is also associated with infection, where it protects against IFNG-mediated hemorrhage. May also facilitate the immune response via both innate and T-cell mediated pathways.
Alternative names
Fibrinogen alpha chain, FGA
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Recommended products
Human Fibrinogen ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human Fibrinogen with a sensitivity of 29 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Key facts
- Detection method
- Fluorescent
- Sample types
- Saliva, Urine, Heparin Plasma, Citrate plasma, Cell culture supernatant, Milk, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 15.63 - 64000 pg/mL
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 29 pg/mL
Precision
Intra assay
| Sample | n | C.V. |
|---|---|---|
Sample Serum | n 8 | C.V. 3.4 |
Inter assay
| Sample | n | C.V. |
|---|---|---|
Sample Serum | n 3 | C.V. 11.7 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Cell culture supernatant | Average % = 94 | Range 93 - 95 % |
Sample type Citrate plasma | Average % = 94 | Range 91 - 96 % |
Sample type EDTA Plasma | Average % = 93 | Range 91 - 94 % |
Sample type Heparin Plasma | Average % = 95 | Range |
Sample type Milk | Average % = 103 | Range 102 - 105 % |
Sample type Saliva | Average % = 101 | Range 98 - 104 % |
Sample type Serum | Average % = 91 | Range 90 - 94 % |
Sample type Urine | Average % = 100 | Range 99 - 100 % |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
Fibrinogen in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Fibrinogen protein in human serum, plasma, urine, saliva, milk, and cell culture supernatant samples.
This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
If using a Molecular Devices' plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.
The CatchPoint SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.
Fibrinogen is a heterohexamer of molecular mass 340 kDa, made up of two sets of alpha, beta, gamma polypeptide chains, and synthesized in the parenchymal cell of the hepatocyte and in the megakaryocyte. Fibrinogen plays a major role in coagulation, and both elevated and decreased levels have clinical significance. Upon cleavage by thrombin in the initial stages of coagulation activation, Fibrinogen self-assembles to yield a fibrin clot matrix that subsequently is crosslinked by factor XIIIa to form an insoluble network. Fibrinogen also binds to the platelet glycoprotein IIb and IIIa receptor so as to form bridges between platelets, thus facilitating aggregation. Elevated plasma Fibrinogen has been identified as an independent risk factor for coronary atherosclerosis and ischemic heart disease. Individuals with congenital absence of Fibrinogen, termed afibrinogenemia, have prolonged bleeding times. Defects in Fibrinogen, alpha are a cause of amyloidosis type 8 (AMYL8) also known as systemic non-neuropathic amyloidosis or Ostertag-type amyloidosis.
CROSS REACTIVITY
The following recombinant proteins were prepared at 50 ng/mL and 8,000 pg/mL and assayed for cross reactivity and no cross reactivity was observed: Human Fibrinogen alpha, Human Fibrinogen beta, Human Fibrinogen gamma, Human Factor XII, Human Plasmin, Human Thrombin, Rat Fibrinogen
The following recombinant proteins were prepared at 50 ng/mL and 8,000 pg/mL and assayed for cross reactivity.
- Human D-Dimer: 27% cross-reactivity
SPECIES REACTIVITY
This kit recognizes human and mouse Fibrinogen protein.
Native purified mouse Fibrinogen was prepared at 8,000 pg/mL and assayed for cross reactivity. 100% cross reactivity was observed.
Other species reactivity was determined by measuring 1:670 diluted serum samples of various species, interpolating the protein concentrations from the human standard curve, and expressing the interpolated concentrations as a percentage of the protein concentration in human serum assayed at the same dilution.
Reactivity < 3% was determined for the following species: Rat
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Sandwich ELISA - Human Fibrinogen ELISA Kit, Fluorescent (ab229419)
Example of human Fibrinogen standard curve in Sample Diluent NS.
The Fibrinogen standard curve was prepared as described in Section 10. Raw data generated on SpectraMax M4 Multi-Mode Microplate Reader is shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human Fibrinogen ELISA Kit, Fluorescent (ab229419)
Interpolated concentrations of native Fibrinogen in human serum and plasma samples.
The concentrations of Fibrinogen were measured in duplicates, interpolated from the Fibrinogen standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:670, plasma (citrate) 1:5x105, plasma (EDTA) 1:5x105, and plasma (heparin) 1:5x105. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibrinogen concentration was determined to be 3.45 μg/mL in serum, 2.86 mg/mL in plasma (citrate), 2.96 mg/mL in plasma (EDTA), and 2.64 mg/mL in plasma (heparin).
Sandwich ELISA - Human Fibrinogen ELISA Kit, Fluorescent (ab229419)
Interpolated concentrations of native Fibrinogen in human urine, saliva, milk and cell culture supernatant samples.
The concentrations of Fibrinogen were measured in duplicates, interpolated from the Fibrinogen standard curves and corrected for sample dilution. Undiluted samples are as follows: urine 1:1.7, saliva 1:133, milk 1:133 and HepG2 supernatant 1:500. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Fibrinogen concentration was determined to be 5.93 ng/mL in urine, 934.5 ng/mL in saliva, 199.7 ng/mL in milk and 1.35 μg/mL in HepG2 supernatant.
Sandwich ELISA - Human Fibrinogen ELISA Kit, Fluorescent (ab229419)
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 Fibrinogen concentration was determined to be 1,070 ng/mL with a range of 647 – 2,027 ng/mL.
Sandwich ELISA - Human Fibrinogen ELISA Kit, Fluorescent (ab229419)
Percent reactivity against related fibrinogen proteins.
The following recombinant/native proteins were prepared at 50 ng/mL and assayed for cross-reactivity. Percent reactivity is determined by interpolating the protein concentrations from the standard curve and expressing the interpolated concentrations as a percentage of the protein concentration assayed at the same concentration.
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