All lanes : Anti-Visfatin antibody (ab45890) at 1/250 dilutionLane 1 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)Lane 2 : Heart (Human) Tissue Lysate - adult normal tissue (ab29431)Lane 3 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue (ab29330)Lysates/proteins at 10 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilutionPerformed under reducing conditions.
Anti-Visfatin antibody (ab45890) at 1/250 dilution + Human Visfatin full length protein (ab52323) at 0.01 µgSecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.Exposure time : 20 seconds
ab45890 staining PBEF in rat brain tissue section by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue from 4% PFA perfused animals underwent overnight fixation in 4% paraformaldehyde, cryoprotected in 30% sucrose and cut using cryostat.The primary antibody was diluted, 1/1000 (PBS + 0.3% Triton X100) and incubated with sample for 18 hours at 20°C. An abcam antibody ab60314, Chromeo488 conjugated goat polyclonal to rabbit IgG was used as secondary, at 1/1000 dilution. This antibody produces a nice staining in rat cortical neurons. Some tracts of fibers were stained as well. The picture shows the staining obtained in cortical neurons.See Abreview
ICC/IF image of ab45890 stained HeLa cells. The cells were 100% methanol fixed (5 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 ab45890 at 10µg/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.