Anti-SHANK3 antibody [N69/46]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SHANK3 antibody [N69/46] (AB193307)

ab193307 stained U20S cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab193307 at 5μg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43μM for 1hour at room temperature.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SHANK3 antibody [N69/46] (AB193307)

ab193307 staining SHANK3 in Rat Primary Neurons DIV14 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal rabbit serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab193307 at 1µg/ml (shown in green) and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control (shown in pseudocolour magenta). Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 100% methanol (5 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• WB

Lab

Western blot - Anti-SHANK3 antibody [N69/46] (AB193307)

All lanes:

Western blot - Anti-SHANK3 antibody [N69/46] (ab193307) at 5 µg/mL

Lane 1:

Cerebellum Mouse Tissue Lysate at 20 µg

Lane 2:

Rat Cortex at 20 µg

Lane 3:

Western blot - Rat brain normal tissue lysate - membrane extract (<a href='/en-us/products/tissue-lysates/rat-brain-normal-tissue-lysate-membrane-extract-ab29473'>ab29473</a>) at 20 µg

Lane 4:

Human Cortex at 20 µg

Lane 5:

Human Striatum Neuronal cell lysate at 20 µg

Lane 6:

Human liver tissue lysate - total protein (<a href='/en-us/products/unavailable/human-liver-tissue-lysate-total-protein-ab29889'>ab29889</a>) at 20 µg

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution

Predicted band size: 186 kDa

Observed band size: 186 kDa

true

Exposure time: 4min