![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.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 232kDa; Plexin B1](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_4/13448_ab90087-203455-IPVI001ab900878m.jpg)
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.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 232kDa; Plexin B1
All lanes : Anti-Plexin B1 antibody (ab90087) at 1/500 dilutionLane 1 : rat brain lysateLane 2 : rat testis lysateLane 3 : fetal kidney lysate
Immunohistochemical staining of formalin-fixed paraffin-embedded rat kidney using ab90087 at 1/100.
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.See Abreview
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.