The human serpin superfamily consists of at least 35 members that target not only serine proteases, but also selected cysteine proteases and non-protease proteins. Serpins bind the protease active site resulting in a major conformational rearrangement that traps the enzyme in a covalent acyl-enzyme intermediate. As protease inhibitors, serpins have an array of functions including regulating blood clotting, the complement pathway, extracellular matrix remodeling, and cell motility. They are also involved in activities that extend beyond their ability to inhibit proteases. For instance, they may also regulate blood pressure, angiogenesis, or act as storage/transport proteins.
Human Serpin A12 Quantikine ELISA Kit
R&D Systems | Catalog # DSA120
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL), Saliva (50 µL), Urine (50 µL), Human Milk (50 µL)
Sensitivity
14.6 pg/mL
Assay Range
31.3-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
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Product Summary for Human Serpin A12 Quantikine ELISA Kit
The Quantikine Human Serpin A12/Vaspin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Serpin A12 in cell culture supernates, serum, plasma, saliva, urine, and human milk. It contains NS0-expressed recombinant human Serpin A12 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Serpin A12 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human Serpin A12.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Human
Specificity
Natural and recombinant human Serpin A12.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Human Milk, Saliva, Serum, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 170 | 531 | 1061 | 173 | 523 | 1033 |
| Standard Deviation | 8.57 | 10.6 | 39.9 | 11.2 | 30.6 | 66.6 |
| CV% | 5.0 | 2.0 | 3.8 | 6.5 | 5.9 | 6.4 |
Recovery for Human Serpin A12 Quantikine ELISA Kit
The recovery of human Serpin A12 spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 98 | 92-107 |
| EDTA Plasma (n=4) | 90 | 80-97 |
| Heparin Plasma (n=4) | 92 | 84-98 |
| Serum (n=4) | 92 | 85-99 |
| Urine (n=4) | 93 | 83-100 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human Serpin A12 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human Serpin A12 Quantikine ELISA Kit
Human Serpin A12 ELISA Standard Curve
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Serpin A12
Alternate Names
OL-64, Vaspin
Gene Symbol
SERPINA12
Additional Serpin A12 Products
Product Documents for Human Serpin A12 Quantikine ELISA Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Serpin A12 Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
Citations for Human Serpin A12 Quantikine ELISA Kit
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Protocols
View specific protocols for Human Serpin A12 Quantikine ELISA Kit (DSA120):
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
100 µL Assay Diluent
50 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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