Mouse Nkx2-5 Antibody (Center)

Anti-Mouse Nkx2-5 Antibody (Center) at 1:1000 dilution + Human heart tissue lysate. Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size: 34 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Anti-Mouse Nkx2-5 Antibody (Center) at 1:1000 dilution + CCRF-CEM whole cell lysate. Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size: 30-42 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 cells labeling Nkx2-5 at 1/25 dilution, followed by Dylight 488-conjugated goat anti-Rabbit IgG secondary antibody at 1/200 dilution (green). Immunofluorescence image showing Nucleus staining on C2C12 cell line. Cytoplasmic actin is detected with Dylight 554 Phalloidin (red). The nuclear counter stain is DAPI (blue).

Immunohistochemical analysis of paraffin-embedded Mouse heart tissue was performed on the Leica BOND RXm. Tissue was fixed with formaldehyde at room temperature, antigen retrieval was by heat mediation with a EDTA buffer (pH9.0). Samples were incubated with primary antibody (1:500) for 1 hours at room temperature. A undiluted biotinylated CRF Anti-Polyvalent HRP Polymer antibody was used as the secondary antibody.

Overlay histogram showing C2C12 cells stained (green line). The cells were fixed with 2% paraformaldehyde and then permeabilized with 90% methanol for 10 min. The cells were then incubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (1:25 dilution) for 60 min at 37°C. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight 488 Conjugated Highly Cross-Adsorbed at 1/200 dilution for 40 min at Room temperature. Isotype control antibody (blue line) was rabbit IgG1 (1 μg/1x10^6 cells) used under the same conditions. Acquisition of > 10000 events was performed.