DENR Antibody

All lanes: Anti-DENR Antibody at 1:1000-1:2000 dilution. Lane 1: A431 whole cell lysate. Lane 2: human skeletal muscle lysate. Lane 3: human brain lysate. Lysates/proteins at 20 µg per lane. Secondary Goat Anti-mouse IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size: 22 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Staining DENR in human skeletal muscle sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.

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

Overlay histogram showing Hela cells stained (green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated 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-Mouse IgG, DyLight 488 Conjugated Highly Cross-Adsorbed at 1/400 dilution for 40 min at 37°C. Isotype control antibody (blue line) was mouse IgG2b (1 μg/1x10^6 cells) used under the same conditions. Acquisition of > 10000 events was performed.