Human PLA2G7/PAF-AH/Lp-PLA2 Quantikine ELISA Kit

  (4 citations)     
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Assay Procedure
Citations (4)
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  • 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 (25 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
  • Sensitivity
    0.284 ng/mL
  • Assay Range
    0.8 - 50 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant human PLA2G7
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC143 , Quantikine Immunoassay Control Set 709 for Human PLA2G7 - Please Inquire
Product Summary
The Quantikine Human PLA2G7 Immunoassay is a 4.5 hour solid phase ELISA designed to measure PLA2G7 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human PLA2G7 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human PLA2G7 showed linear curves that were parallel to the 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 PLA2G7.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 1.69 3.52 11.7 2.35 4.34 13
Standard Deviation 0.115 0.192 0.272 0.226 0.3 0.676
CV% 6.8 5.5 2.3 9.6 6.9 5.2

Recovery

The recovery of PLA2G7 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 92 85-105
EDTA Plasma (n=4) 101 91-112
Heparin Plasma (n=4) 102 92-106
Serum (n=4) 106 91-115
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of PLA2G7 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
 PLA2G7/PAF-AH/Lp-PLA2 [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: PLA2G7/PAF-AH/Lp-PLA2
Secretory phopholipase A2 (PLA2) enzymes cleave an acyl ester bond in the sn-2 position of glycerophospholipids. These extracellular proteins have a high disulfide bond content, low molecular mass (14 kDa), and require mM levels of Ca2+ for catalysis. They play a crucial role in the generation of arachidonates and eicosanoids, and have a number of biological actions including immunological responses, inflammation, cellular proliferation, vasoconstriction, and bronchioconstriction.
  • Long Name:
    Phospholipase A2 Group VII
  • Entrez Gene IDs:
    7941 (Human); 27226 (Mouse); 301265 (Rat)
  • Alternate Names:
    2-acetyl-1-alkylglycerophosphocholine esterase; EC 3.1.1; EC 3.1.1.47,1-alkyl-2-acetylglycerophosphocholine esterase; Group-VIIA phospholipase A2; gVIIA-PLA2; LDL-associated phospholipase A2; LDL-PLA(2); LDL-PLA2; lipoprotein-associated phospholipase A2; LpPLA2; Lp-PLA2; PAF acetylhydrolase; PAF-AH; PAFAHPAF 2-acylhydrolase; phospholipase A2, group VII (platelet-activating factor acetylhydrolase; PLA2G7; plasma); platelet-activating factor acetylhydrolase
Related Research Areas
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.
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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Species
Sample Type
  1. Myeloperoxidase level and inflammatory markers and lipid and lipoprotein parameters in stable coronary artery disease
    Authors: E Kimak, B Zi?ba, D Duma, J Solski
    Lipids Health Dis, 2018;17(1):71.
    Species: Human
    Sample Type: Serum
  2. A transcriptome-based global map of signaling pathways in the ovarian cancer microenvironment associated with clinical outcome
    Genome Biol, 2016;17(1):108.
    Species: Human
    Sample Type: Ascites Fluid
  3. Impact of cigarette smoking cessation on high-density lipoprotein functionality.
    Authors: Takata K, Imaizumi S, Kawachi E, Suematsu Y, Shimizu T, Abe S, Matsuo Y, Tsukahara H, Noda K, Yahiro E, Zhang B, Uehara Y, Miura S, Saku K
    Circ J, 2014;78(12):2955-62.
    Species: Human
    Sample Type: Plasma
  4. Plasma desmoplakin I biomarker of vascular recurrence after ischemic stroke.
    Authors: Lopez-Farre AJ, Zamorano-Leon JJ, Segura A, Mateos-Caceres PJ, Modrego J, Rodriguez-Sierra P, Calatrava L, Tamargo J, Macaya C
    J. Neurochem., 2012;121(2):314-25.
    Species: Human
    Sample Type: Plasma

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ELISA Controls
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Quantikine Immunoassay Control Set 709 for Human PLA2G7

Ctrl QC143
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Supplemental ELISA Products
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Quantikine ELISA Wash Buffer 1

ELISA WA126 6  
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