Human SP-D Quantikine ELISA Kit

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DSFPD0
Human SP-D ELISA Cell Culture Supernate Standard Curve
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Product Details
Procedure
Citations (7)
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Reviews (1)

Human SP-D Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
5.5 hours
Sensitivity
0.37 ng/mL
Assay Range
0.6 - 40 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human SP-D
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.

Product Summary

The Quantikine Human SP-D Immunoassay is a 5.5 hour solid-phase ELISA designed to measure human SP-D in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human SP-D and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human SP-D 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 ocurring SP-D.

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

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (ng/mL) 6.07 11.7 23.4 5.63 11.2 22.8
Standard Deviation 0.495 0.588 1.46 0.495 0.999 1.78
CV% 8.2 5 6.2 8.8 8.9 7.8

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (ng/mL) 5.99 11.3 24.3 6.63 13.1 26.9
Standard Deviation 0.371 0.921 1.63 0.617 1.14 2.5
CV% 6.2 8.2 6.7 9.3 8.7 9.3

Recovery

The recovery of 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) 102 95-109
EDTA Plasma (n=4) 110 99-120
Heparin Plasma (n=4) 104 85-118
Serum (n=4) 106 89-118

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of SP-D were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human SP-D ELISA Linearity

Scientific Data

Human SP-D ELISA Cell Culture Supernate Standard Curve

Human SP-D ELISA Serum/Plasma 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: 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
⚠ 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 3 hours.
  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.
  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. 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 for Human SP-D 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.

7 Citations: Showing 1 - 7
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  1. Biomarkers of inflammation and epithelial barrier function in multiple sclerosis
    Authors: A Olsson, S Gustavsen, IC Hasselbalc, AR Langkilde, F Sellebjerg, AB Oturai, HB Søndergaar
    Mult Scler Relat Disord, 2020;46(0):102520.
    Species: Human
    Sample Types: Serum
  2. Improving risk-stratification of rheumatoid arthritis patients for interstitial lung disease
    Authors: J Avouac, A Cauvet, A Steelandt, Y Shirai, M Elhai, M Kuwana, O Distler, Y Allanore
    PLoS ONE, 2020;15(5):e0232978.
    Species: Human
    Sample Types: Serum
  3. Club cell protein 16 and cytokeratin fragment 21-1 as early predictors of pulmonary complications in polytraumatized patients with severe chest trauma
    Authors: LL Negrin, G Halat, S Kettner, M Gregori, R Ristl, S Hajdu, T Heinz
    PLoS ONE, 2017;12(4):e0175303.
    Species: Human
    Sample Types: Serum
  4. Circulating thymus and activation-regulated chemokine/CC chemokine ligand 17 is a strong candidate diagnostic marker for interstitial lung disease in patients with malignant tumors: a result from a pilot study.
    Authors: Yamane H, Ochi N, Yamagishi T, Honda Y, Takeyama M, Takigawa N
    Ther Clin Risk Manag, 2015;11(0):949-59.
    Species: Human
    Sample Types: Serum
  5. Intraoperative cell salvage during cardiac surgery is associated with reduced postoperative lung injury.
    Authors: Engels G, van Klarenbosch J, Gu Y, van Oeveren W, de Vries A
    Interact Cardiovasc Thorac Surg, 2015;22(3):298-304.
    Species: Human
    Sample Types: Plasma
  6. Expansive generation of functional airway epithelium from human embryonic stem cells.
    Authors: McIntyre B, Alev C, Mechael R, Salci K, Lee J, Fiebig-Comyn A, Guezguez B, Wu Y, Sheng G, Bhatia M
    Stem Cells Transl Med, 2014;3(1):7-17.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Nintedanib modulates surfactant protein-D expression in A549 human lung epithelial cells via the c-Jun N-terminal kinase-activator protein-1 pathway.
    Authors: Kamio K, Usuki J, Azuma A, Matsuda K, Ishii T, Inomata M, Hayashi H, Kokuho N, Fujita K, Saito Y, Miya T, Gemma A
    Pulm Pharmacol Ther, 0;32(0):29-36.
    Species: Human
    Sample Types: Cell Lysates

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Human SP-D Quantikine ELISA Kit
By Anonymous on 10/04/2020
Sample Tested: Serum

This assay passed our internal qualification with good dilutional linearity, accuracy, and precision.