Tubulin, Sigma-Aldrich, T5076

Sigma # T5076: mouse-anti-β-III-tub

The antibody was used for Immunofluorescence staining of neurons derived from murine ES cells differentiated in culture.
The cells were grown on coated cover slips, fixed for 30min in 4% PFA and treated for 5 min with 0,005% Triton-X-100. To reduce unspecific binding cells are blocked/ quenched with a 1% BSA in PBS solution.
The Antibody was diluted 1:8000. The secondary antibody was a goat anti mouse 488 diluted 1:1000.
Strong staining of the neurons was observed.
In double staining experiments of the same cell lines nearly no β-III-tub could be observed when mouse-anti-β-III-tub is used together with Millipore # AB152 rabbit-anti-TH (1:500) or Sigma # A2052 rabbit-anti-GABA (1:250).

Please see inserted pictures with the staining of Sigma # T5076 mouse-anti-β-III-tub The pictures show the differentiated wildtype cells from the ES cell line CJ7.