Human HMW Kininogen ELISA Kit, Fluorescent (ab229438)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 9.5 pg/ml
- Range: 9.8 pg/ml - 20000 pg/ml
- Sample type: Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Saliva, Serum, Urine
- Detection method: Fluorescent
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human HMW Kininogen ELISA Kit, Fluorescent -
Detection method
Fluorescent -
Precision
Intra-assay Sample n Mean SD CV% Serum 8 3.9% Inter-assay Sample n Mean SD CV% Serum 3 11.4% -
Sample type
Cell culture supernatant, Saliva, Urine, Serum, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
9.5 pg/ml -
Range
9.8 pg/ml - 20000 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture supernatant 107 99% - 111% Saliva 117 114% - 120% Urine 118 115% - 120% Serum 89 88% - 90% Hep Plasma 90 84% - 93% EDTA Plasma 90 87% - 92% Cit plasma 88 82% - 91% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human
Does not react with: Cow -
Product overview
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.
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Notes
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.
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Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 100X Stoplight Red Substrate 1 x 120µl 10X Human HMW Kininogen Capture Antibody 1 x 600µl 10X Human HMW Kininogen Detector Antibody 2 vials 10X Wash Buffer PT (ab206977) 1 x 20ml 500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl Antibody Diluent 5BI 1 x 6ml Human HMW Kininogen Lyophilized Purified Protein (ab90353) 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated Black 96-Well Microplate 1 unit Stoplight Red Substrate Buffer 1 x 12ml -
Research areas
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Function
(1) Kininogens are inhibitors of thiol proteases; (2) HMW-kininogen plays an important role in blood coagulation by helping to position optimally prekallikrein and factor XI next to factor XII; (3) HMW-kininogen inhibits the thrombin- and plasmin-induced aggregation of thrombocytes; (4) the active peptide bradykinin that is released from HMW-kininogen shows a variety of physiological effects: (4A) influence in smooth muscle contraction, (4B) induction of hypotension, (4C) natriuresis and diuresis, (4D) decrease in blood glucose level, (4E) it is a mediator of inflammation and causes (4E1) increase in vascular permeability, (4E2) stimulation of nociceptors (4E3) release of other mediators of inflammation (e.g. prostaglandins), (4F) it has a cardioprotective effect (directly via bradykinin action, indirectly via endothelium-derived relaxing factor action); (5) LMW-kininogen inhibits the aggregation of thrombocytes; (6) LMW-kininogen is in contrast to HMW-kininogen not involved in blood clotting. -
Tissue specificity
Secreted in plasma. T-kinin is detected in malignant ovarian, colon and breast carcinomas, but not in benign tumors. -
Involvement in disease
Defects in KNG1 are the cause of high molecular weight kininogen deficiency (HMWK deficiency) [MIM:228960]. HMWK deficiency is an autosomal recessive coagulation defect. Patients with HWMK deficiency do not have a hemorrhagic tendency, but they exhibit abnormal surface-mediated activation of fibrinolysis. -
Sequence similarities
Contains 3 cystatin domains. -
Post-translational
modificationsBradykinin is released from kininogen by plasma kallikrein.
Hydroxylation of Pro-383 occurs prior to the release of bradykinin.
Phosphorylation sites are present in the extracelllular medium.
N- and O-glycosylated. O-glycosylated with core 1 or possibly core 8 glycans. -
Cellular localization
Secreted > extracellular space. - Information by UniProt
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Alternative names
- Alpha 2 thiol proteinase inhibitor
- Alpha-2-thiol proteinase inhibitor
- BDK
see all -
Database links
- Entrez Gene: 3827 Human
- Omim: 612358 Human
- SwissProt: P01042 Human
- Unigene: 77741 Human
Images
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Other - Human HMW Kininogen ELISA Kit, Fluorescent (ab229438)
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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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.
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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).
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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.
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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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Sandwich ELISA - Human HMW Kininogen ELISA Kit, Fluorescent (ab229438)To learn more about the advantages of recombinant antibodies see here.
