Description
Principle of the Assay::The Rat Neuropilin 2 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Rat Neuropilin 2 in Cell Culture Supernatants, Cell Lysates, Tissue Homogenates. This assay employs an antibody specific for Rat Neuropilin 2 coated on a 96-well plate. Standards and samples are pipetted into the wells and Neuropilin 2 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Rat Neuropilin 2 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Neuropilin 2 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Background/Introduction: NRP2 (Neuropilin 2), also called Npn2 or VEGF165R2, encodes a member of the neuropilin family of receptor proteins. The soluble NRP2 was secreted as a 62.5-kD protein following transfection in Chinese hamster ovary cells. The NRP2 gene is mapped to 2q33.3. The NRP2 gene contains 17 exons and spans about 112 kb. This protein may play a role in cardiovascular development, axon guidance, and tumorigenesis. Mice with null mutations in genes encoding Sema3F, and its holoreceptor components Npn2 and plexin A3 (PLEXA3), exhibit increased dentate gyrus granule cell and cortical layer V pyramidal neuron spine number and size, and also aberrant spine distribution. Moreover, Sema3F promotes loss of spines and excitatory synapses in dissociated neurons in vitro, and in NRP2-null brain slices cortical layer V and dentate gyrus granule cells exhibit increased miniature excitatory postsynaptic current frequency. These disparate effects of secreted semaphorins are reflected in the restricted dendritic localization of NRP2 to apical dendrites and of Npn1 to all dendrites of cortical pyramidal neurons