Western blot - Anti-GAP43 antibody [EP890Y] (ab75810)
All lanes : Anti-GAP43 antibody [EP890Y] (ab75810) at 1/20000 dilution
Lane 1 : Mouse brain tissue lysate Lane 2 : Rat brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) at 1/1000 dilution
Predicted band size : 25 kDa
Exposure time : 3 minutes
Blocking Buffer: 5% NFDM/TBST
Diluting Buffer: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - GAP43 antibody [EP890Y] (ab75810)This image is a courtesy of anonymous Abreview.
ab75810 staining GAP43 in Mouse ear tissue sections by Immunohistochemistry (Formalin/ PFA-fixed paraffin-embedded tissue sections). The sections were formaldehyde fixed, subjected to heat mediated antigen retrieval at pH 6 and blocked for 10 minutes at 25C. The primary antibody was diluted 1/500 and incubated with the sample for 1 hour at 25°C. An HRP polymer anti-rabbit IgG system was used undiluted, as the secondary antibody.
Overlay histogram showing SH-SY5Y cells stained with ab75810 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75810, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
ICC/IF image of ab75810 stained SKNSH 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 (ab75810, 1/50 dilution) 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.
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