Human SPA (Pulmonary Surfactant Associated Protein A) ELISA Kit (HUES02324)
- Product Type:
- ELISA Kit
- 96 Assays
- ELISA Type:
- MGC133365, PSAP, PSPA, SFTP1, SFTPA1
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cell Biology
|Detection Range:||15.63-1000 pg/mL|
|Sample Volume Required Per Well:||100µL|
|Sample Type:||Serum, plasma and other biological fluids|
|Specificity:||This kit recognizes Human SPA in samples. No significant cross-reactivity or interference between Human SPA and analogues was observed.|
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human SPA. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human SPA and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human SPA, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human SPA. The concentration of Human SPA in samples can be calculated by comparing the OD of the samples to the standard curve.
|UniProt Protein Function:||SFTPA1B: In presence of calcium ions, it binds to surfactant phospholipids and contributes to lower the surface tension at the air-liquid interface in the alveoli of the mammalian lung and is essential for normal respiration. Genetic variations in SFTPA1 are a cause of susceptibility to pulmonary fibrosis idiopathic (IPF). Pulmonary fibrosis is a lung disease characterized by shortness of breath, radiographically evident diffuse pulmonary infiltrates, and varying degrees of inflammation and fibrosis on biopsy. It results in acute lung injury with subsequent scarring and endstage lung disease. Genetic variations in SFTPA1 are a cause of susceptibility to respiratory distress syndrome in premature infants (RDS); also known as RDS in prematurity. RDS is a lung disease affecting usually premature newborn infants. It is characterized by deficient gas exchange, diffuse atelectasis, high-permeability lung edema and fibrin-rich alveolar deposits called 'hyaline membranes'. The association between SFTPA1 alleles and respiratory distress syndrome in premature infants is dependent on a variation Ile to Thr at position 131 in SFTPB. Belongs to the SFTPA family.|
|UniProt Protein Details:|
Protein type:Secreted; Secreted, signal peptide
Chromosomal Location of Human Ortholog: 10q22. 3
Cellular Component: extracellular space; proteinaceous extracellular matrix; collagen; extracellular region
Molecular Function:lipid transporter activity; carbohydrate binding
Biological Process: respiratory gaseous exchange; lipid transport
Disease: Pulmonary Fibrosis, Idiopathic
|NCBI Summary:||This gene encodes a lung surfactant protein that is a member of a subfamily of C-type lectins called collectins. The encoded protein binds specific carbohydrate moieties found on lipids and on the surface of microorganisms. This protein plays an essential role in surfactant homeostasis and in the defense against respiratory pathogens. Mutations in this gene are associated with idiopathic pulmonary fibrosis. Alternate splicing results in multiple transcript variants. [provided by RefSeq, May 2010]|
|NCBI GenInfo Identifier:||60416440|
|NCBI Gene ID:||653509|
|NCBI Accession:||Q8IWL2. 2|
|UniProt Secondary Accession:||Q8IWL2,P07714, Q14DV4, Q5RIR5, Q5RIR7, A8K3T8, B7ZW50 E3VLD8, E3VLD9, E3VLE0, E3VLE1, G5E9J3,|
|UniProt Related Accession:||Q8IWL2|
|Molecular Weight:||27,736 Da|
|NCBI Full Name:||Pulmonary surfactant-associated protein A1|
|NCBI Synonym Full Names:||surfactant protein A1|
|NCBI Official Symbol:||SFTPA1|
|NCBI Official Synonym Symbols:||SPA; PSAP; PSPA; SP-A; SPA1; PSP-A; SFTP1; SP-A1; COLEC4; SFTPA1B|
|NCBI Protein Information:||pulmonary surfactant-associated protein A1; collectin-4; surfactant protein A1B; alveolar proteinosis protein; surfactant protein A1 variant AD' 6A; surfactant protein A1 variant ACD' 6A; surfactant protein A1 variant AD' 6A2; surfactant protein A1 variant AD' 6A3; surfactant protein A1 variant AD' 6A4; surfactant protein A1 variant AB'D' 6A; surfactant protein A1 variant ACD' 6A2; surfactant protein A1 variant ACD' 6A3; surfactant protein A1 variant ACD' 6A4; surfactant protein A1 variant AB'D' 6A2; surfactant protein A1 variant AB'D' 6A3; surfactant protein A1 variant AB'D' 6A4; surfactant, pulmonary-associated protein A1A; surfactant, pulmonary-associated protein A1B; 35 kDa pulmonary surfactant-associated protein|
|UniProt Protein Name:||Pulmonary surfactant-associated protein A1|
|UniProt Synonym Protein Names:||35 kDa pulmonary surfactant-associated protein; Alveolar proteinosis protein; Collectin-4|
|Protein Family:||Pulmonary surfactant-associated protein|
|UniProt Gene Name:||SFTPA1|
|UniProt Entry Name:||SFTA1_HUMAN|
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human SPA were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human SPA were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||5.01||5.59||3.90||6.01||4.36||4.72|
The recovery of Human SPA spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||85-98||91|
|Cell culture media (n=5)||88-101||93|
Samples were spiked with high concentrations of Human SPA and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
|Serum (n=5)||EDTA plasma (n=5)||Cell culture media (n=5)|
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
|Micro ELISA Plate(Dismountable)||8 wells ×12 strips||-20°C, 6 months|
|Reference Standard||2 vials|
|Concentrated Biotinylated Detection Ab (100×)||1 vial, 120 µL|
|Concentrated HRP Conjugate (100×)||1 vial, 120 µL||-20°C(shading light), 6 months|
|Reference Standard & Sample Diluent||1 vial, 20 mL||4°C, 6 months|
|Biotinylated Detection Ab Diluent||1 vial, 14 mL|
|HRP Conjugate Diluent||1 vial, 14 mL|
|Concentrated Wash Buffer (25×)||1 vial, 30 mL|
|Substrate Reagent||1 vial, 10 mL||4°C(shading light)|
|Stop Solution||1 vial, 10 mL||4°C|
|Plate Sealer||5 pieces|
|Product Description||1 copy|
|Certificate of Analysis||1 copy|
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100 µL of standard solutions into the standard wells.
- Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
- Add 100 µL of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37 °C.
- Aspirate the liquid from each well, do not wash. Immediately add 100 µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37 °C.
- Aspirate or decant the solution from the plate and add 350 µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100 µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37 °C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 90 µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37 °C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.