Mouse SP-D Quantikine ELISA Kit

  (1 citations)     
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Assay Procedure
Citations (1)
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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 (50 uL), Tissue Lysates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
  • Sensitivity
    9.43 pg/mL
  • Assay Range
    62.5 - 4,000 pg/mL (Cell Culture Supernates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant mouse SP-D.
  • 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 SP-D immunoassay is a 4.5 hour solid-phase ELISA designed to measure SP-D in mouse cell culture supernates, tissue lysates, serum, and plasma. It contains CHO cell-expressed recombinant mouse SP-D and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant mouse SP-D. Results obtained using natural mouse SP-D showed dose response curves that were parallel to the standard curves obtained using the Quantikine mouse kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse SP-D.

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 kit components
Cell Culture Supernates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 4 4 4 4 4 4
Mean 158 534 1270 169 531 1267
Standard Deviation 5.75 13.8 43 14 34.4 120
CV% 3.6 2.6 3.4 8.3 6.5 9.5

Recovery

The recovery of mouse SP-D spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 99 92-105
EDTA Plasma (n=4) 97 90-103
Heparin Plasma (n=4) 97 84-104
Serum (n=4) 96 84-109
Tissue Lysates (n=4) 101 91-113
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse SP-D in each matrix were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay
 SP-D [HRP]
Product Datasheets

Certificate of Analysis Lookup

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: SP-D
Surfactant Protein D (SP-D) is a collectin family protein consisting of a collagen-like sequence followed by a C-type lectin domain. It is synthesized primarily in the lung by alveolar type II cells and unciliated bronchial epithelial cells. In addition, SP-D is produced by epithelial cells of the gastrointestinal tract and exocrine glands. SP-D binds to patterns of neutral sugars on micro-organisms to mediate direct opsonization and phagocytosis and is an important component of the innate immune system.
  • Long Name:
    Surfactant Pulmonary Associated Protein D
  • Entrez Gene IDs:
    6441 (Human); 20390 (Mouse)
  • Alternate Names:
    COLEC7collectin-7; Collectin 7; Collectin-7; Lung surfactant protein D; PSPD; PSP-D; SFTP4; SFTP4pulmonary surfactant-associated protein D; SFTPD; SPD; SP-D; SP-Dpulmonary surfactant apoprotein; surfactant protein D; surfactant, pulmonary-associated protein D; surfactant-associated protein, pulmonary 4
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. 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 3 times for a total of 4 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 4 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:

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.

1 Citations: Showing 1 - 1

  1. Stat5 Is Required for CD103(+) Dendritic Cell and Alveolar Macrophage Development and Protection from Lung Injury
    Authors: WE Eddy, KQ Gong, B Bell, WC Parks, SF Ziegler, AM Manicone
    J. Immunol., 2017;0(0):.
    Species: Mouse
    Sample Type: BALF

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