Santa Cruz Biotechnology, Inc.
α-tubulin antibody
sc-32293
DM1A
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To find a loading control, tissues were collected and lysed with the buffer [1% SDS, 10 mmol/L Tris-Cl (pH 7.6), 20 μg/μL aprotinin, 20 μg/μL leupeptin, and 1 mmol/L 4-(2-aminoethyl)benzenosulfonyl fluoride]. The protein concentrations were determined using the Bicinchoninic Acid Protein Assay kit (Pierce, Rockford, IL). Fiften micrograms of protein were separated on SDS-PAGE gels and transferred to polyvinylidene difluoride membranes. After blocking, the membranes were incubated with the appropriately diluted primary antibody (1/1000) at 4°C overnight. After washing with TBST, the membranes were incubated with goat anti-mouse diluted HRP-conjugated secondary antibody (1/10000) at room temperature for 1 hour. Proteins were detected with the enhanced chemiluminescence kit (Amersham Pharmacia Biotechnology, Inc., Piscataway, NJ).
Western Blot
Mouse brain and spleen
1/1000 at 4°C overnight
5% milk
goat anti-mouse diluted HRP-conjugated secondary antibody (1/10000)
room temperature for 1 hour
the enhanced chemiluminescence kit (Amersham Pharmacia Biotechnology, Inc., Piscataway, NJ)
The results showed that the α-tubulin was a good loading control in mouse brain (Left) and spleen (Right).
The antibody works well
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