ELISA Kit for Surfactant Protein B (SP-B) Homo sapiens (Human) Sandwich ELISA

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

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  • ELISA Kit for Surfactant Protein B (SP-B) Packages (Simulation)
  • ELISA Kit for Surfactant Protein B (SP-B) Packages (Simulation)
  • ELISA Kit for Surfactant Protein B (SP-B) Results demonstration
  • SEB622Hu.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) and the recovery rates were calculated by comparing the measured value to the expected amount of Surfactant Protein B (SP-B) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 99-105 102
EDTA plasma(n=5) 85-104 94
heparin plasma(n=5) 82-105 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Surfactant Protein B (SP-B) 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) 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) 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) 79-98% 79-103% 97-105% 80-102%
EDTA plasma(n=5) 98-105% 88-95% 95-102% 81-102%
heparin plasma(n=5) 78-94% 89-96% 82-104% 95-104%

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.

ELISA Kit for Surfactant Protein B (SP-B)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Surfactant Protein B (SP-B). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Surfactant Protein B (SP-B). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Surfactant Protein B (SP-B), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Surfactant Protein B (SP-B) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

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

Citations

  • s**********b@**dizin.uni-leipzig.de
    August 24, 2018
    Elevated Surfactant Protein Levels and Increased Flow of Cerebrospinal Fluid in Cranial Magnetic Resonance Imaging.
    Molecular Neurobiology

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