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CELSR2 was immunoprecipitated using 0.5mg Mouse Brain tissue lysate, 5µg of Rabbit polyclonal to CELSR2 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 Brain tissue lysate 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 ab90817.
Anti-CELSR2 antibody (ab90817) at 1 µg/ml + Human brain tissue lysate - total protein (ab29466) at 10 µg
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 317 kDa Observed band size : 317 kDa
Exposure time : 16 minutesAbcam recommends loading unheated lysates and using 20% Ethanol and 0.04% SDS in the transfer buffer to obtain a positive signal for this antibody. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above
ICC/IF image of ab90817 stained SKNSH 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 ab90817 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.
References for Anti-CELSR2 antibody (ab90817)
has not yet been referenced specifically in any publications.
Publishing research using ab90817? Please let us know so that we can cite the reference in this datasheet.
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