All lanes : Anti-Aquaporin 3 antibody (ab125219) at 1 µg/mlLane 1 : Rat kidney tissue lysateLane 2 : Mouse lung tissue lysateLane 3 : Mouse kidney tissue lysate
ab125219 at 1µg/ml staining Aquaporin 3 in Paraaffin-embedded Rat kidney tissue by Immunohistochemistry.
Aquaporin 3 was immunoprecipitated using 0.5mg Mouse Kidney extract, 5µg of Rabbit polyclonal to Aquaporin 3 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, Mouse Kidney 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab125219.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 31kDa: Aquaporin 3.
ICC/IF image of ab125219 stained MCF-7 cells. The cells were 4% formaldehyde fixed (10 min) 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 ab125219 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 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.