Anti-Oct4 antibody - ChIP Grade (ab19857)

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 (1:10000) -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 (1:10000) -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

Protein of tissues and cells was extracted using a RIPA lysis buffer kit (R0010, Solarbio, Beijing, China). Protein concentration was measured by the bicinchoninic acid (BCA) method (69-21875, MSK Biological Technological Co., Ltd., Wuhan, China). After 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE, 20050227, Beyotime Biotechnology, Haimen, Jiangsu, China), the membrane was transferred and underwent antigen-antibody reaction. Primary antibodies including Notch antibody (1:75, ab27526), CTNNB1 antibody (1:5000, ab32572), Hes1 antibody (1:50, ab71559), BIM antibody (1:2000, ab32158), Bax antibody (1:2000, ab32503), Bcl-2 antibody (1:2000, ab32124), Runx2 antibody (1:10000, ab76956), osteocalcin antibody (1:10000, ab19857), and GAPDH antibody (1:1000, ab9485) which were purchased from Abcam (Cambridge, MA, USA) were added to the membrane and incubated at 4°C overnight. The membrane was then incubated with HRP-labeled goat-anti-rabbit IgG secondary antibody (1:1000, BA1056, BOSTER Biological Technology Co., Ltd., Wuhan, Hubei, China) and then immersed in electrochemiluminescence (ECL, WBKLS0500, Pierce, Rockford., IL, U.S.A.). The result was observed after developing the color in a dark room and photographs were taken. GAPDH was used as the internal protein reference and the relative protein expression level was presented as the gray value ratio of target band to the internal reference band.

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Manufacturer protocol

Download the product protocol from Abcam for Anti-Oct4 antibody - ChIP Grade (ab19857) below.

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