Multiplex Assay Kit for Surfactant Associated Protein D (SPD) ,etc. by FLIA (Flow Luminescence Immunoassay)

SFTPD; COLEC7; PSP-D; SFTP4; SP-D; Pulmonary Surfactant Protein D; Collectin-7; Lung surfactant protein D

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Surfactant Associated Protein D (SPD) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Surfactant Associated Protein D (SPD) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Surfactant Associated Protein D (SPD) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Surfactant Associated Protein D (SPD) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 92-99 95
EDTA plasma(n=5) 86-94 90
heparin plasma(n=5) 81-99 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Surfactant Associated Protein D (SPD) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Surfactant Associated Protein D (SPD) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Surfactant Associated Protein D (SPD) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 87-94% 96-103% 87-102% 96-103%
EDTA plasma(n=5) 85-97% 80-97% 79-104% 84-92%
heparin plasma(n=5) 87-102% 94-103% 91-98% 79-97%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:SPD) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
The American Journal of Rhinology & Allergy Surfactant protein A and D in chronic rhinosinusitis with nasal polyposis and corticosteroid response Ingenta: art00005
J Occup Health Evaluation of acute oxidative stress induced by NiO nanoparticles in vivo and in vitro PubMed: 21233593
Inhalation Toxicology Comparison of acute oxidative stress on rat lung induced by nano and fine-scale, soluble and insoluble metal oxide particles: NiO and TiO2 PubMed: 22642288
American Journal of Rhinology & Allergy Detection of surfactant proteins A, B, C, and D in human nasal mucosa and their regulation in chronic rhinosinusitis with polyps PubMed: 23406594
Plos one Staphylococcus aureus and Pseudomonas aeruginosa Express and Secrete Human Surfactant Proteins PubMed: PMC3551896
Plos one The Detection of Surfactant Proteins A, B, C and D in the Human Brain and Their Regulation in Cerebral Infarction, Autoimmune Conditions and Infections of the CNS PubMed: PMC3787032
Nachweis und Charakterisierung des Oberfl?chenproteins PLUNC (Palate, Lung and Nasal Clone Protein) an der Augenoberfl?che und Bedeutung für das Trockene Auge Opus4:Source
PLoS One The Cerebral Surfactant System and Its Alteration in HydrocephalicConditions. pubmed:27656877
Frontiers in Aging Neuroscience Correlations of Ventricular Enlargement with Rheologically Active SurfactantProteins in Cerebrospinal Fluid. pubmed:28101052
American Journal of Physiology. Lung Cellular and Molecular Physiology Chronic lung injury and impaired pulmonary function in a mouse model of acid ceramidase deficiency. pubmed:29167126
Molecular Neurobiology Rheologically Essential Surfactant Proteins of the CSF Interacting with Periventricular White Matter Changes in Hydrocephalus Patients–Implications for CSF … Doi: 10.1007/s12035-019-01648-z
Respir Res Elevated plasma levels of epithelial and endothelial cell markers in COVID-19 survivors with reduced lung diffusing capacity six months after hospital … Pubmed:35189887
Virology Journal The association between sPD-1 levels versus liver biochemistry and viral markers in chronic hepatitis B patients: a comparative study of different sPD-1 … Pubmed:35361235
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