Western blotting Jak1

Western blotting is a widely used technique to size separate proteins from a pool of cell or tissue lysates. The technique has 4 major steps: a) gel electrophoresis, b) blocking and treatment with antigen specific antibody, c) treatment with secondary antibody and finally d) detection and visualization. Though western blotting is a widely used technique, detection of specific proteins depends on several factors, the major ones are antibody concentration, incubation time and washing steps. Key points for obtaining clean blots are: always prepare fresh buffer solutions and optimize antibody concentration. Given the advent of high-throughput protein analysis and a push to limit the use of lab consumables, onestep antibodies are developed which recognise protein of interest and also contain a detection label.

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Found 3 matching solutions for this experiment

Upstream tips
-These buffers may be stored at 4°C for several weeks or aliquoted and stored at -20°C for up to a year.
Protocol tips
-Rabbit (1:500)
-To reduce and denature your samples, boil each cell lysate in sample buffer at 100°C for
5 min. Lysates can be aliquoted and stored at -20°C for future use.
JAK1 Antibody (B-3): sc-376996

Santa Cruz Biotechnology

Protocol tips
-Mouse (1:500)
-Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink
can fluoresce and cause background.
Protocol tips
-Rabbit (1:1000)
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