TTC36 Antibody / Tetratricopeptide repeat protein 36 (FY13091)

Immunofluorescent staining of TTC36 using anti-TTC36 antibody (green) and anti-Beta Tubulin antibody (red). TTC36 was detected in immunocytochemical section of cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/ml rabbit anti-TTC36 antibody and mouse anti-Beta Tubulin antibody overnight at 4oC. DyLight 488 Conjugated Goat Anti-Rabbit IgG and Cy3 Conjugated Goat Anti-Mouse IgG were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. Visualize using a fluorescence microscope and filter sets appropriate for the label used. TTC36 antibody (green) shows nuclear-enriched staining with nucleolar sparing, consistent with reported nuclear localization. Beta-tubulin antibody (red) marks the cytoplasmic microtubule network surrounding the nucleus.

Western blot analysis of TTC36 using anti-TTC36 antibody. Lane 1: mouse liver tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-TTC36 antibody at 0.5 ug/ml overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using enhanced chemiluminescent. A strong band is detected at ~21 kDa, matching the expected size of TTC36. A faint ~28 kDa band is also present and likely represents a modified or isoform variant of TTC36 (e.g., mono-ubiquitinated or glycosylated species), which is variably observed in tissue lysates.

Flow Cytometry analysis of 293T cells using anti-TTC36 antibody. Overlay histogram showing 293T cells stained with (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TTC36 antibody (1 ug/million cells) for 30 min at 20oC. DyLight 488 conjugated goat anti-rabbit IgG (5-10 ug/million cells) was used as secondary antibody for 30 minutes at 20oC. Isotype control antibody (Green line) was rabbit IgG (1 ug/million cells) used under the same conditions. Unlabelled sample (Red line) was also used as a control.