Mouse Pentraxin 3/TSG-14 Quantikine ELISA Kit

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MPTX30
Mouse Pentraxin 3/TSG-14 Standard Curve
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Mouse Pentraxin 3/TSG-14 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 (13 uL), EDTA Plasma (13 uL), Heparin Plasma (13 uL)
Sensitivity
0.02 ng/mL
Assay Range
0.2 - 15 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant mouse PTX3
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 Mouse Pentraxin 3 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse PTX3 in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse PTX3 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse PTX3 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 mouse PTX3.

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 33 34 34
Mean 0.54 2.09 6.86 0.63 2.28 7.18
Standard Deviation 0.01 0.05 0.2 0.07 0.16 0.41
CV% 1.9 2.4 2.9 11.1 7 5.7

Recovery

The recovery of mouse PTX3 spiked to three levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 107 98-119
EDTA Plasma (n=4) 98 90-110
Heparin Plasma (n=4) 95 81-100
Serum (n=4) 102 90-117

Linearity

To assess the linearity of the assay, samples containing or spiked with high concentrations of mouse PTX3 in each matrix were diluted with Calibrator Diluent and assayed.
Mouse Pentraxin 3/TSG-14 ELISA Linearity

Product Datasheets

Preparation and Storage

Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Pentraxin 3/TSG-14

Pentraxin 3 (PTX3), also known as TNF-stimulated gene 14 (TSG-14), is a secreted glycoprotein belonging to the Pentraxin family of proteins. It is the prototypical long Pentraxin, exhibiting a C-terminal Pentraxin domain characteristic of the family, and a unique N-terminal domain. It is produced by several cell types including endothelial and mononuclear cells. PTX3 acts as a pattern recognition receptor with non-redundant roles in the innate immune response to several microbial agents including the fungal pathogen Aspergillus fumigatus. Overexpression leads to enhanced pro-inflammatory responses. PTX3 is induced in response to LPS and inflammatory cytokines including TNF-alpha and IL-1 beta.

Long Name:
Tumor Necrosis Factor Stimulated Gene 14
Entrez Gene IDs:
5806 (Human); 19288 (Mouse)
Alternate Names:
alpha-induced protein 5; pentaxin-related gene, rapidly induced by IL-1 beta, tumor necrosis factor; Pentaxin-related protein PTX3; Pentraxin 3; pentraxin 3, long; pentraxin-3; pentraxin-related gene, rapidly induced by IL-1 beta; pentraxin-related protein PTX3; PTX3; TNF alpha-induced protein 5; TNFAIP5; TSG14; TSG-14; TSG-14pentaxin-related gene, rapidly induced by IL-1 beta; Tumor necrosis factor alpha-induced protein 5; tumor necrosis factor, alpha-induced protein 5; Tumor necrosis factor-inducible gene 14 protein; tumor necrosis factor-inducible protein TSG-14
⚠ 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. 50 µL Assay Diluent
  4.   Add 50 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 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 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 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Mouse Pentraxin 3/TSG-14 Quantikine ELISA Kit

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.

3 Citations: Showing 1 - 3
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  1. MicroRNA-224 is involved in the regulation of mouse cumulus expansion by targeting Ptx3.
    Authors: Yao G, Liang M, Liang N, Yin M, Lu M, Lian J, Wang Y, Sun F
    Mol Cell Endocrinol, 2014;382(1):244-53.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. Endothelial pentraxin 3 contributes to murine ischemic acute kidney injury.
    Authors: Chen, Jianlin, Matzuk, Martin M, Zhou, Xin J, Lu, Christop
    Kidney Int, 2012;82(11):1195-207.
    Species: Mouse
    Sample Types: Plasma
  3. Protective effects of long pentraxin PTX3 on lung injury in a severe acute respiratory syndrome model in mice.
    Authors: Han, Bing, Ma, Xuezhong, Zhang, Jianhua, Zhang, Yu, Bai, Xiaohui, Hwang, David M, Keshavjee, Shaf, Levy, Gary A, McGilvray, Ian, Liu, Mingyao
    Lab Invest, 2012;92(9):1285-96.
    Species: Mouse
    Sample Types: Serum

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