Plexin B1 is the transmembrane receptor for the class 4 semaphorin, SEMA4D. Plexin B1 activation plays a role in axon guidance, invasive growth and cell migration and has been reported to have additional roles in RHOA activation and subsequent changes of the actin cytoskeleton.
Plexin B1 was immunoprecipitated using 0.5mg Rat brain tissue extract, 5µg of Rabbit polyclonal to Plexin B1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Rat brain tissue extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab90087.
Immunohistochemical staining of formalin-fixed paraffin-embedded rat kidney using ab90087 at 1/100.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Plexin B1 antibody (ab90087)This image is courtesy of an Abreview submitted by Manoj Kumar Valluru
ab90087 staining Plexin B1 in Mouse brain tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% serum for 1 hour at room temperature; antigen retrieval was by heat mediation in citrate buffer (pH 6). Samples were incubated with primary antibody (1/500 in PBS + 2% Blocking Serum) for 16 hours at 4°C. A Biotin-conjugated goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.
ICC/IF image of ab90087 stained PC12 cells. The cells were 4% PFA fixed (10mins) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab90087, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.