MTT was purchased from Sigma-Aldrich; Merck Millipore (Darmstadt, Germany). Polyclonal rabbit anti-human cleaved caspase-3 (1:1,000; cat. no. 9661S), monoclonal rabbit anti-human cleaved caspase-8 (1:1,000; cat. no. 9496S), polyclonal rabbit anti-human cleaved caspase-9 (1:1,000; cat. no. 9505S), monoclonal mouse anti-human BCL-2 (1:1,000; cat. no. 15071S), polyclonal rabbit anti-human Bax (1:1,000; cat. no. 2772S), monoclonal rabbit anti-human cyclin D1 (1:1,000; cat. no. 2978S), monoclonal rabbit anti-human CDK4 (1:1,000; cat. no. 12790S), monoclonal rabbit anti-human CDK6 (1:1,000; cat. no. 13331S) and monoclonal rabbit anti-human p21 (1:1,000; cat. no. 2947S) primary antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). N-Benzyloxycarbonyl-Val-Ala-Asp (O-Me) fluoromethyl ketone (Z-VAD-FMK) was purchased from Beyotime Institute of Biotechnology (Haimen, China). The monoclonal mouse anti-human β-actin primary antibody was obtained from Abcam (1:1,000; cat. no. ab8226; Cambridge, UK). Goat anti-mouse and goat anti-rabbit secondary antibodies were purchased from Thermo Fisher Scientific, Inc., (1:5,000; cat. nos. A16072 and A16110, respectively; Waltham, MA, USA).
Monoclonal mouse anti human β actin primary antibody
Manufactured by Abcam
Sourced in United Kingdom
Monoclonal mouse anti-human β-actin primary antibody is a laboratory reagent designed for the detection and quantification of β-actin, a ubiquitous cellular protein, in biological samples. This antibody is produced in mice and specifically recognizes the human β-actin protein.
Automatically generated - may contain errors
2 protocols using monoclonal mouse anti human β actin primary antibody
1
Apoptosis Marker Antibody Protocol
2
Western Blot Analysis of RIZ1 Expression
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The TE13 cells transfected with pcDNA3.1(+)/RIZ1 were homogenized in radioimmunoprecipitation buffer (containing 50 mm Tris-HCl, pH 7.4; 150 mm NaCl; 1% Nonidet P-40; 0.5% sodium deoxycholate; 0.1% SDS; 1 mm EDTA; 1 mm phenylmethylsulfonyl fluoride and 1 mg/ml aprotinin) and the protein concentrations were determined using a bicinchoninic acid protein assay kit (Pierce, Rockford, IL, USA). Cell lysates (30 μg) were separated by 8% SDS-PAGE, transferred to nitrocellulose membranes (Amersham Biosciences, Chalfont St. Giles, UK) and immunoblotted with the indicated antibodies overnight in the Orbital Shaker (Thermo Fisher Scientific, Waltham, MA, USA) at 4°C. The monoclonal mouse anti-human RIZ1/PR domain antibodies, monoclonal mouse anti-human β-actin primary antibody and secondary polyclonal goat anti-mouse polyclonal antibody were all obtained from Abcam (Cambridge, UK).
Bands were visualized using a PowerLook scanner (UMAX Technologies, Hsinchu, Taiwan) and quantified using ImageQuant software (GE Healthcare, Little Chalfont, UK). The relative expression levels of RIZ1 and the PR domain were calculated as the gray values of RIZ1, the PR domain and β-actin. Untransfected and empty vector-transfected TE13 cells served as negative controls.
Bands were visualized using a PowerLook scanner (UMAX Technologies, Hsinchu, Taiwan) and quantified using ImageQuant software (GE Healthcare, Little Chalfont, UK). The relative expression levels of RIZ1 and the PR domain were calculated as the gray values of RIZ1, the PR domain and β-actin. Untransfected and empty vector-transfected TE13 cells served as negative controls.
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