Anti-Zic2 抗体 [EPR7790] (ab150404)

Diluting/Blocking buffer and concentration: 5% NFDM/TBST

It detects an 70kDa extra band in some cell lines.

Immunocytochemistry analysis of SK-OV-3 (human ovarian cancer epithelial cell)cells labelling Zic2 with Ab150404 at 2.5 μg/ml. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 was used as the secondary antibody (green). Cells were counterstained with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red). Nuclear DNA was labelled with DAPI (blue).

Exposure time:

Lane1: 3.25 seconds
Lane2: 20 seconds
Lane3: 5.5 seconds
Lane4-5: 60 seconds

Diluting/Blocking buffer and concentration: 5% NFDM/TBST

ab150404 detects an 70kDa extra band in some cell lines. The expression profile observed in U-87 MG is consistent with the literature (PMID: 11038359). Negative control: U-87 MG (PMID: 11038359)

Intracellular Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Zic2 with purified ab150404 at 1/600 dilution (1µg/ml) (red). Cells were fixed with 80% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). 

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Overlay histogram showing SH-SY5Y cells stained with unpurified ab150404 (red line). The cells were fixed with 80% methanol (5 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 (unpurified ab150404, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.