Anti-Bcl-2 antibody (ab196495)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-These buffers may be stored at 4°C for several weeks or aliquoted and stored at -20°C for up to a year.	-Rabbit (1:1200) -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.

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 (1:1200) -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.

Publication protocol

Proteins of SVZ area and neural stem cells were prepared using RIPA buffer followed by high-speed centrifugation (12000 rpm, 20 min) at 4°C. After collecting the supernatants, the protein concentration was determined by BCA kit, and 40 μg of total proteins were isolated with 10% SDS/PAGE. The targeted proteins were transferred on to nitrocellulose membranes (Millipore, Billerica, MA, U.S.A.) and then incubated with 5% low-fat dried milk for 2 h to block the non-specific binding sites. The primary antibodies were used to incubate with membranes at 4°C overnight. After washing with PBS twice, the membranes were incubated with horseradish peroxidase–conjugated secondary antibodies for 2 h, and then the signals were detected by enhanced chemiluminescent reagents. The antibodies in the present study included TAOK1 (Rabbit, 1:1000, ab197891, Abcam), GAPDH (1:3000, sc420485, Santa Cruz), AKT (Rabbit, 1:1000, ab106693, Abcam), p-AKT (Rabbit, 1:1000, ab38449, Abcam), Bax (Rabbit, 1:1200, ab263897, Abcam), Bcl-2 (Rabbit, 1:1200, ab196495, Abcam), Cleaved-caspase-3 (Rabbit, 1:1000, ab2302, Abcam), CyclinD1 (goat, 1:1500, ab194972, Abcam), extracellular signal-regulated kinase (ERK) (Rabbit, 1:1000, ab137619, Abcam), p-ERK (Rabbit, 1:1000, ab79483, Abcam), P-21 (Mouse, 1:1000, ab168220, Abcam), PI3K (Rabbit, 1:1200, ab70912, Abcam), P38 (Rabbit, 1:1000, ab27986, Abcam), p-P38 (Rabbit, 1:1000, ab47363, Abcam).

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Manufacturer protocol

Download the product protocol from Abcam for Anti-Bcl-2 antibody (ab196495) below.

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