Human Serpin A12 Quantikine ELISA Kit

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DSA120
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Human Serpin A12 ELISA Standard Curve
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Human Serpin A12 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Saliva (50 uL), Urine (50 uL), Human Milk (50 uL)
Sensitivity
14.6 pg/mL
Assay Range
31.3 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
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.

Product Summary

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.

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, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk

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 2 3.8 6.5 5.9 6.4

Recovery

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.
Human Serpin A12 ELISA Linearity

Scientific Data

Human Serpin A12 ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Serpin A12

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.

Entrez Gene IDs:
145264 (Human); 68054 (Mouse); 191570 (Rat)
Alternate Names:
antitrypsin), member 12; member 12; OL-64; OL-64Visceral adipose-specific serpin; serine (or cysteine) proteinase inhibitor, clade A (alpha-1 antiproteinase; Serpin A12; serpin peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin); Vaspin; Visceral adipose tissue-derived serine protease inhibitor
⚠ 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.

Assay Procedure

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.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   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.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   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.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   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.

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