Anti-alpha Tubulin antibody [TU-16] (ab7749)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - alpha Tubulin antibody [TU-16] (ab7749)

Ab7749 staining human normal skin. Staining is localised to the cytoplasm.
Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

  Immunohistochemistry (Frozen sections) - alpha Tubulin antibody [TU-16] (ab7749)

ab7749 at 1/500 dilution staining alpha tubulin in chicken embryo trunk section by Immunohistochemistry (Frozen sections). Embryonic trunks were electroporated at Hamburger and Hamilton (HH) stages 14-16 with mem-EGFP, harvested 16-19 hours after electroporation at HH stages 17-19 and cryosectioned at 30 μm. Cryosections were rehydrated in PBS, the cells permeabilized in -20°C acetone for 20 minutes, washed in PBS and then blocked in PBS containing 2% normal goat serum (1% BSA, 0.1% cold water fish skin gelatin, 0.1% Triton X-100 and 0.05% Tween 20) for 20 minutes. Blocking solution was removed and cryosections were incubated overnight with primary antibody at 4°C. A Cy5 conjugated goat anti mouse IgM at 1/300 dilution was used as secondary.

Image from Ahlstrom JD et.al., Development 136, 1801-1812 (2009), Fig 7B.