Human HMW Kininogen ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human HMW Kininogen with a sensitivity of 9.5 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, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 9.8 - 20000 pg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 9.5 pg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Target data
Function
Kininogens are inhibitors of thiol proteases. HMW-kininogen plays an important role in blood coagulation by helping to position optimally prekallikrein and factor XI next to factor XII; HMW-kininogen inhibits the thrombin- and plasmin-induced aggregation of thrombocytes. LMW-kininogen inhibits the aggregation of thrombocytes. LMW-kininogen is in contrast to HMW-kininogen not involved in blood clotting. Bradykinin. The active peptide bradykinin is a potent vasodilatator that is released from HMW-kininogen shows a variety of physiological effects: (A) influence in smooth muscle contraction, (B) induction of hypotension, (C) natriuresis and diuresis, (D) decrease in blood glucose level, (E) it is a mediator of inflammation and causes (E1) increase in vascular permeability, (E2) stimulation of nociceptors (4E3) release of other mediators of inflammation (e.g. prostaglandins), (F) it has a cardioprotective effect (directly via bradykinin action, indirectly via endothelium-derived relaxing factor action).
Alternative names
BDK, KNG, KNG1, Kininogen-1, Alpha-2-thiol proteinase inhibitor, Fitzgerald factor, High molecular weight kininogen, Williams-Fitzgerald-Flaujeac factor, HMWK
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Human HMW Kininogen ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human HMW Kininogen with a sensitivity of 9.5 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, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 9.8 - 20000 pg/mL
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 9.5 pg/mL
Precision
Intra assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Serum | n 8 | mean - | SD - | C.V. 3.9 |
Inter assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Serum | n 3 | mean - | SD - | C.V. 11.4 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Cell culture supernatant | Average % = 107 | Range 99 - 111 % |
Sample type Serum | Average % = 89 | Range 88 - 90 % |
Sample type EDTA Plasma | Average % = 90 | Range 87 - 92 % |
Sample type Saliva | Average % = 117 | Range 114 - 120 % |
Sample type Urine | Average % = 118 | Range 115 - 120 % |
Sample type Heparin Plasma | Average % = 90 | Range 84 - 93 % |
Sample type Citrate plasma | Average % = 88 | Range 82 - 91 % |
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
HMW Kininogen in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of HMW Kininogen protein in human serum, plasma, and cell culture supernatants.
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.
High molecular weight kininogen (HMW kininogen) is a 72kDa highly glycosylated protein with an important role in the assembly of the plasma kallikrein-kinin system and in the blood coagulation process. It is encoded by the KNG1 gene, which generates both HMWK and low molecular weight kininogen (LMW kininogen) via alternative splicing. Both HMW kininogen and LMW kininogen share a heavy chain consisting of protein domains 1, 2 and 3 and differ in their light chain. HMW kininogen contains a 56kDa light chain consisting of domain 5 and 6H, whereas LMW kininogen contains a 4kDa light chain consisting of domain 5L. Heavy and light chains of HMW kininogen and LMW kininogen are linked via domain 4 which contains the bradykinin nonapeptide.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
High molecular weight kininogen also known as HMW kininogen or HK is a glycoprotein with a molecular mass of about 120 kDa. It is synthesized in the liver and circulates in the bloodstream. It serves as a precursor to bradykinin a peptide involved in inflammation blood pressure regulation and pain. HMW kininogen also acts as a cofactor for kallikrein activation playing a significant role in the intrinsic pathway of blood coagulation. Additionally it exists as part of a quaternary structure with other plasma proteins functioning within the circulatory system.
- Biological function summary
This kininogen is essential for the kallikrein-kinin system and interacts with various cell surface receptors to trigger diverse cellular responses. It is part of a complex that includes prekallikrein and factor XII facilitating the assembly and activation of these components on negatively charged surfaces. This interaction supports the generation of bradykinin which mediates vasodilation and increases vascular permeability critical processes in inflammatory responses. The multifunctional nature of HMW kininogen allows it to contribute broadly to the regulation of hemostasis and innate immunity.
- Pathways
HMW kininogen is linked notably to both the contact activation pathway and the renin-angiotensin system. In the contact activation pathway it associates closely with prekallikrein and factor XII promoting the conversion of prekallikrein to kallikrein and subsequent steps in coagulation. In the renin-angiotensin system bradykinin produced from HMW kininogen modulates blood pressure through interactions with ACE (angiotensin-converting enzyme) showcasing the enzyme's relationship with these pathways. These pathways position HMW kininogen at a critical junction of cardiovascular and immune system regulation.
- Associated diseases and disorders
HMW kininogen associates with conditions such as hereditary angioedema and cardiovascular disorders. Hereditary angioedema results from bradykinin overproduction implicating HMW kininogen in its pathophysiology with links to defects in C1 inhibitor another regulatory protein. Cardiovascular disorders may involve aberrant bradykinin production and metabolism where HMW kininogen plays a role through interaction with the renin-angiotensin system. Aberrations in its function or expression can contribute toward the pathogenesis of these conditions highlighting the need for further research into its clinical implications.
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4 product images
Sandwich ELISA - Human HMW Kininogen ELISA Kit, Fluorescent (ab229438)
Example of human HMW Kininogen standard curve in Sample Diluent NS.
The HMW Kininogen 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 HMW Kininogen ELISA Kit, Fluorescent (ab229438)
Interpolated concentrations of native HMW Kininogen in human serum and plasma samples.
The concentrations of HMW Kininogen were measured in duplicates, interpolated from the HMW Kininogen standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:80,000, plasma (citrate) 1:320,000, plasma (heparin) 1:320,000 and plasma (EDTA) 1:80,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean HMW Kininogen concentration was determined to be 163 μg/mL in serum, 202 μg/mL in plasma (citrate), 151 μg/mL in plasma (EDTA), and 173 μg/mL in plasma (heparin).
Sandwich ELISA - Human HMW Kininogen ELISA Kit, Fluorescent (ab229438)
Interpolated concentrations of native HMW Kininogen in human urine, saliva, and HepG2 cell supernatant samples.
The concentrations of HMW Kininogen were measured in duplicates, interpolated from the HMW Kininogen standard curves and corrected for sample dilution. Undiluted samples are as follows: urine 5%, saliva 50%, and HepG2 cell supernatant 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean HMW Kininogen concentration was determined to be 38,903 pg/mL in urine, 2,857 pg/mL in saliva, and 7,454 pg/mL in HepG2 cell supernatant.
Sandwich ELISA - Human HMW Kininogen ELISA Kit, Fluorescent (ab229438)
Serum from eight individual healthy human female donors was measured in duplicate.
Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean HMW Kininogen concentration was determined to be 79 μg/mL with a range of 62-112 μg/mL.
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