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Mouse SP-D ELISA Kit - Quantikine

Catalog #: MSFPD0
4 Citations 2 Reviews Datasheet / COA / SDS
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MSFPD0
Mouse SP-D ELISA Standard Curve
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Product Details
Procedure
Citations (4)
FAQs
Supplemental Products
Reviews (2)
Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Mouse SP-D 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), 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 (pg/mL) 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
Mouse SP-D ELISA Linearity

Scientific Data

N/A Mouse SP-D ELISA Standard Curve View Larger

Mouse SP-D 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: 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
&#9888; 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
  3.   Add 50 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   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.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 100 µL Conjugate
  6.   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.
  7.   Aspirate and wash 4 times.
    8. 100 µL Substrate Solution
  8.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    9. 100 µL Stop Solution
  9.   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 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.

4 Citations: Showing 1 - 4
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  1. NETs Promote Inflammatory Injury by Activating cGAS-STING Pathway in Acute Lung Injury
    Authors: J Zhao, N Zhen, Q Zhou, J Lou, W Cui, G Zhang, B Tian
    International Journal of Molecular Sciences, 2023-03-07;24(6):.
    Species: Mouse
    Sample Types: Serum
  2. Change of surfactant protein D and A after renal ischemia reperfusion injury
    Authors: IM Imtiazul, R Asma, JH Lee, NJ Cho, S Park, HY Song, HW Gil
    PLoS ONE, 2019-12-26;14(12):e0227097.
    Species: Mouse
    Sample Types: Serum
  3. Macrophage migration inhibitory factor enhances influenza-associated mortality in mice
    Authors: CA Smith, DJ Tyrell, UA Kulkarni, S Wood, L Leng, RL Zemans, R Bucala, DR Goldstein
    JCI Insight, 2019-07-11;4(13):.
    Species: Mouse
    Sample Types: Serum
  4. 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., 2017-05-12;0(0):.
    Species: Mouse
    Sample Types: BALF

FAQs

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Reviews for Mouse SP-D Quantikine ELISA Kit

Average Rating: 4.5 (Based on 2 Reviews)

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Mouse SP-D Quantikine ELISA Kit
By Anonymous on 02/18/2021
Sample Tested: Serum
Mouse SP-D Quantikine ELISA Kit MSFPD0
Mouse SP-D Quantikine ELISA Kit
By Anonymous on 03/06/2019
Sample Tested: FVBN mouse plasma and BAL

I used this kit to analyze SP-D of FVBN mice plasma and BAL infected with 0111:B4 LPS. I had a lot of background signal which made it hard to distinguish the difference of SP-D between control and infected mice. I think the kit itself is good and easy to use though, I just need to increase the difference to compensate for background noise. I also didn't dilute enough in my BAL, so my std curve was not allowed to develop as much as I would have liked. If you run with BAL I would recommend a 1:500 dilution! I did 1:200.

Mouse SP-D Quantikine ELISA Kit MSFPD0