Santa Cruz Biotechnology
β Tubulin
sc-5274
D-10
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Cells were isolated from albino wistar rats and extract was prepared. The extract was centrifuged and the supernatant was collected. Protein was estimated in the supernatant and 40 microgram of the protein was loaded in wells and resolved in the SDS-PAGE. The protein bands were separated and then transferred to the nitrocellulose membrane by performing electroblotting. Nitrocellulose membrane was blocked by skim milk and then incubated overnight with primary antibody (β Tubulin). Membrane was washed and then incubated with secondary antibody conjugated with HRP. The ECL reagent is used for producing the signal which is captured by x ray film.
Western Blot
Mammalian cell
12 h incubation at 4°C (1:1000 dilution in 5% BSA prepared in TBST)
5% blotting grade blocker (skimmed milk) in TBST
1 h at room temperature (1:4000 dilution in 5% BSA prepared in TBST)
None
ECL reagent
Specific bands were observed with the β Tubulin antibody. All the samples exhibited similar bands as β Tubulin is a housekeeping protein.
N/A
The antibodies were specific and gave sharp bands.
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