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Figure 2 | Breast Cancer Research

Figure 2

From: β class II tubulin predominates in normal and tumor breast tissues

Figure 2

Western blots of normal and tumor breast tissues after sample enrichment for tubulin by arginine–Sepharose chromatography. For quantitative Western blots the linearity of response for each antibody was established for PC-tubulin. A linear response was also determined for the antibody reaction with purified tubulin isotype samples. It was not necessary to know total protein levels in these experiments, because percentage isotypes were being quantified. (A) Immunoblot with monoclonal antibody 7B9, specific for β-tubulin class II. Lane 1, 3.9 μg of PC-tubulin; lane 2, 0.35 μg of purified β-tubulin class II; lane 3, 30 μl of normal tissue sample, lane 4, 30 μl of tumor tissue sample. (B) Immunoblot with monoclonal antibody TUJ1, specific for β-tubulin class III. Lane 1, 3.9 μg of PC-tubulin; lane 2, 0.23 μg of purified β-tubulin class III; lane 3, 30 μl of normal tissue sample; lane 4, 30 μl of tumor tissue sample. (C) Immunoblot with monoclonal antibody 7H12, specific for β-tubulin classes I + IV. Lane 1, 3.9 μg of PC-tubulin; lane 3, 30 μl of normal tissue sample; lane 4, 30 μl of tumor tissue sample. Note that because β-tubulin classes I + IV constitute only about 17% of the total tubulin in pig brain, we did not purify sufficient quantities for controls on these Western blots. We determined the quantity of β-tubulin classes I + IV by subtraction.

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