Wide-range ELISA Kit for Surfactant Associated Protein D (SPD)

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Wide-range Surfactant Associated Protein D (SPD).
No significant cross-reactivity or interference between Wide-range Surfactant Associated Protein D (SPD) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Wide-range Surfactant Associated Protein D (SPD) and the recovery rates were calculated by comparing the measured value to the expected amount of Wide-range Surfactant Associated Protein D (SPD) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 94-105 99
EDTA plasma(n=5) 80-99 91
heparin plasma(n=5) 78-91 84

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Wide-range Surfactant Associated Protein D (SPD) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Wide-range Surfactant Associated Protein D (SPD) 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 Wide-range Surfactant Associated Protein D (SPD) 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) 85-98% 97-104% 89-102% 98-105%
EDTA plasma(n=5) 95-104% 95-104% 96-104% 97-104%
heparin plasma(n=5) 80-97% 96-103% 81-97% 80-90%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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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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