Multiplex Assay Kit for Pulmonary Surfactant Associated Protein A1 (SFTPA1) ,etc. by FLIA (Flow Luminescence Immunoassay) Canis familiaris; Canine (Dog) Multiplex ELISA

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

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

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  • Multiplex Assay Kit for Pulmonary Surfactant Associated Protein A1 (SFTPA1) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Pulmonary Surfactant Associated Protein A1 (SFTPA1) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Pulmonary Surfactant Associated Protein A1 (SFTPA1) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA890Ca.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Pulmonary Surfactant Associated Protein A1 (SFTPA1) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Pulmonary Surfactant Associated Protein A1 (SFTPA1) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 88-97 93
EDTA plasma(n=5) 96-103 101
heparin plasma(n=5) 97-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Pulmonary Surfactant Associated Protein A1 (SFTPA1) ,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 Pulmonary Surfactant Associated Protein A1 (SFTPA1) ,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 Pulmonary Surfactant Associated Protein A1 (SFTPA1) ,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) 95-102% 83-94% 83-98% 87-95%
EDTA plasma(n=5) 78-97% 93-101% 85-104% 80-97%
heparin plasma(n=5) 96-105% 98-105% 81-92% 94-102%

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#:SFTPA1) 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.

Multiplex Assay Kit for Pulmonary Surfactant Associated Protein A1 (SFTPA1) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer.The MFI developed is proportional to the concentration of analytes of interest in the sample.

Giveaways

Citations

  • Surfactant protein A and D in chronic rhinosinusitis with nasal polyposis and corticosteroid responseIngenta: art00005
  • Detection of surfactant proteins A, B, C, and D in human nasal mucosa and their regulation in chronic rhinosinusitis with polypsPubMed: 23406594
  • Staphylococcus aureus and Pseudomonas aeruginosa Express and Secrete Human Surfactant ProteinsPubMed: PMC3551896
  • 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 CNSPubMed: PMC3787032
  • Nachweis und Charakterisierung des Oberfl?chenproteins PLUNC (Palate, Lung and Nasal Clone Protein) an der Augenoberfl?che und Bedeutung für das Trockene AugeOpus4:Source
  • Expression of surfactant protein-A in exhaled breath condensate of patients with chronic obstructive pulmonary diseasePubMed: 26707652
  • Serum Levels of Surfactant Proteins in Patients with Combined Pulmonary Fibrosis and Emphysema (CPFE)Pubmed:27337142
  • The Cerebral Surfactant System and Its Alteration in HydrocephalicConditions.pubmed:27656877
  • Correlations of Ventricular Enlargement with Rheologically Active SurfactantProteins in Cerebrospinal Fluid.pubmed:28101052
  • Correlations of Ventricular Enlargement with Rheologically Active Surfactant Proteins in Cerebrospinal FluidPMC5209370
  • Chronic lung injury and impaired pulmonary function in a mouse model of acid ceramidase deficiency.pubmed:29167126
  • Metabolic profiling of asthma in mice and the interventional effects of SPA using liquid chromatography and Q-TOF mass spectrometrypubmed:28463380
  • Evaluation of surfactant proteins A, B, C, and D in articular cartilage, synovial membrane and synovial fluid of healthy as well as patients with osteoarthritis and …Pubmed: 30235245
  • Organic barn dust inhibits surfactant protein D production through protein kinase-c alpha dependent increase of GPR116Pubmed: 30543664
  • Elevated levels of multifunctional surfactant proteins in cerebrospinal fluid are associated with signs of increased cerebrospinal fluid flow in cranial magnetic …Doi: 10.1007/s12035-017-0835-5
  • 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
  • Oxidative Stress and Inflammatory Mediators in Exhaled Breath Condensate of Patients with Pulmonary Tuberculosis. A Pilot Study with a Biomarker Perspective34679707

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