Anti-SHANK2 antibody [N23B/49]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SHANK2 antibody [N23B/49] (AB94575)

4% formaldehyde-fixed (for 15 minutes at room temperature) SK-N-BE (Human neuroblastoma cells) labeling SHANK2 using ab94575 at 1/100 dilution (for 60 minutes at room temperature). Goat Anti-Mouse ATTO 488 was used as the secondary at 1/200 dilution for 60 minutes at room temperature. Localization in the sytplasm is observed.

Counterstains : Phalloidin Texas Red F-Actin stain (Red). DAPI nuclear stain (Blue).

• WB

Lab

Western blot - Anti-SHANK2 antibody [N23B/49] (AB94575)

Western blot : Anti-SHANK2 antibody [N23B/49] (ab94575) staining at 1 ug/ml, shown in green; Rabbit anti-ACTN4 [EPR2533(2)] (ab108198) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab94575 was shown to bind specifically to SHANK2. A band was observed at 201 kDa in wild-type A549 cell lysates with no signal observed at this size in SHANK2 knockout cell line. To generate this image, wild-type and SHANK2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-SHANK2 antibody [N23B/49] (ab94575) at 1 µg/mL

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

SHANK2 knockout A549 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

Predicted band size: 158 kDa

Observed band size: 201 kDa

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• WB

Unknown

Western blot - Anti-SHANK2 antibody [N23B/49] (AB94575)

Western blot detection of SHANK2 in Rat braim membrane lysates using ab94575 at 1 : 1000 dilution.

All lanes:

Western blot - Anti-SHANK2 antibody [N23B/49] (ab94575)

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