Multiplex Assay Kit for Surfactant Protein B (SP-B) ,etc. by FLIA (Flow Luminescence Immunoassay) Rattus norvegicus (Rat) Multiplex ELISA

SFTPB; PSPB; SFTB3; SFTP3; SPB; Surfactant Associated Protein B; Pulmonary Surfactant Protein B; 18 kDa pulmonary-surfactant protein; 6 kDa protein

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

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  • Multiplex Assay Kit for Surfactant Protein B (SP-B) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Surfactant Protein B (SP-B) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Surfactant Protein B (SP-B) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMB622Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 79-90 84
EDTA plasma(n=5) 87-99 95
heparin plasma(n=5) 90-98 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Surfactant Protein B (SP-B) ,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 Protein B (SP-B) ,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 Protein B (SP-B) ,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) 80-97% 80-92% 82-96% 87-94%
EDTA plasma(n=5) 93-104% 89-96% 78-95% 95-104%
heparin plasma(n=5) 93-101% 96-104% 97-104% 82-93%

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#:SP-B) 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 Surfactant Protein B (SP-B) ,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

  • Kinetics of plasma SPB and RAGE during mechanical ventilation in patients undergoing major vascular surgeryPubMed: 21736957
  • Detection of surfactant proteins A, B, C, and D in human nasal mucosa and their regulation in chronic rhinosinusitis with polypsPubMed: 23406594
  • Surfactant protein B and RAGE increases in the plasma during cardiopulmonary bypass: a pilot studyPubMed: 20650982
  • 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
  • Lung Cancer Signatures in Plasma Based on Proteome Profiling of Mouse Tumor ModelsPubMed: PMC3406925
  • Acute high-altitude exposure reduces lung diffusion: Data from the HIGHCARE Alps projectPubmed: 23619193
  • Surfactant-Derived Proteins as Markers of Alveolar Membrane Damage in Heart FailurePubmed:25514679
  • Nachweis und Charakterisierung des Oberfl?chenproteins PLUNC (Palate, Lung and Nasal Clone Protein) an der Augenoberfl?che und Bedeutung für das Trockene AugeOpus4:Source
  • Plasma immature form of surfactant protein type B correlates with prognosis in patients with chronic heart failure. A pilot single-center prospective studyPubMed: 26310985
  • 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
  • Diving and pulmonary physiology: surfactant binding protein, lung fluid and cardiopulmonary test changes in professional diverspubmed:28467885
  • Surfactant Protein B Suppresses Lung Cancer Progression by Inhibiting Secretory Phospholipase A2 Activity and Arachidonic Acid Productionpubmed:28743125
  • Chronic lung injury and impaired pulmonary function in a mouse model of acid ceramidase deficiency.pubmed:29167126
  • Effects and molecular mechanisms of intrauterine infection/inflammation on lung developmentPubmed:29747649
  • Surfactant proteins changes after acute hemodynamic improvement in patients with advanced chronic heart failure treated with LevosimendanPubmed:29548887
  • Serum Surfactant Protein Levels in Patients Admitted to the Hospital with Acute COPD ExacerbationPubmed:29445934
  • Generation of an alveolar epithelial type II cell line from induced pluripotent stem cellsPubmed: 30211653
  • Culture of human alveolar epithelial type II cells by sproutingPubmed: 30340591

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