Surfactant, pulmonary-associated protein D, also known as SFTPD or SP-D, is a protein which in humans is encoded by the SFTPD gene. By fluorescence in situ hybridization, the SP-D gene was localized in 10q22.2-q23.1. On the basis of homology with other collectins, potential functions for SP-D include roles in innate immunity and surfactant metabolism, SP-D is produced in the bronchiolar and terminal epithelium of human fetal lung from about 21 weeks of gestation. SP-A and SP-D act as dual-function surveillance molecules that reverse orientation and function and become initiators of host-defense reactions.
Sample Type:Cell culture supernates, serum and plasma (heparin, EDTA)
Intended Use:For quantitative detection of human SP-D in cell culture supernates, serum and plasma (heparin, EDTA).
Sensitivity:<20pg/ml
Range:625-40,000pg/ml
Specificity:Natural and recombinant human SP-D
Test Principle:This human SP-D BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for SP-D has been precoated onto 96-well plates. Standards(NSO) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for SP-D is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human SP-D amount of sample captured in plate.
Kit Components:96-well plate precoated with human SP-D antibody, 1Lyophilized recombinant human SP-D standard, 40ng/tube×2Biotinylated human SP-D antibody, 130ul (dilution 1:100)Avidin-Biotin-Peroxidase Complex (ABC), 130ul (dilution 1:100)Sample diluent buffer, 30mlAntibody diluent buffer, 12mlABC diluent buffer, 12mlTMB color developing agent, 10mlTMB stop solution, 10ml
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
