Wide-range ELISA Kit for Surfactant Associated Protein A (SPA)

PRL; SFTPA; PSAP; PSPA; SFTP; SFTPA1; COLEC4; PSP-A; SFTP1; SFTPA1B; SP-A1; Collectin-4; Alveolar proteinosis protein; 35 kDa pulmonary surfactant-associated protein

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 81-94 87
EDTA plasma(n=5) 87-103 99
heparin plasma(n=5) 78-104 96

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Wide-range Surfactant Associated Protein A (SPA) 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 A (SPA) 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 A (SPA) 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) 98-105% 91-98% 79-104% 78-96%
EDTA plasma(n=5) 83-103% 94-102% 91-98% 90-98%
heparin plasma(n=5) 86-105% 91-105% 81-99% 99-105%

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
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
Mol Med Rep Expression of surfactant protein-A in exhaled breath condensate of patients with chronic obstructive pulmonary disease PubMed: 26707652
Plos One Serum Levels of Surfactant Proteins in Patients with Combined Pulmonary Fibrosis and Emphysema (CPFE) Pubmed:27337142
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
Frontiers in Aging Neuroscience Correlations of Ventricular Enlargement with Rheologically Active Surfactant Proteins in Cerebrospinal Fluid PMC5209370
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 biosystems Metabolic profiling of asthma in mice and the interventional effects of SPA using liquid chromatography and Q-TOF mass spectrometry pubmed:28463380
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