Immunoblotting procedure was as described previously (42 (link)). Primary antibodies include: ELL3 (1:300 dilution) (#H000080237-B02P lot WuLz 08310, and H00080237-B01P lot E1172, 08295 WuLz, Abnova, Taipei City, Taiwan), ELL2 (1:10,000 dilution) (A302-505A; Bethyl Laboratories Inc., Montgomery, TX), ELL (1:800 dilution) (#51044-1-AP, Proteintech Group, Chicago, IL), β-actin (1:12,000 dilution) (AC-15, Sigma Aldrich, St. Louis, MO), PARP (46D11), Phospho-Histone H2A.X (Ser139) (#2577), Cleaved Caspase-3 (Asp175) (#9661), Cyclin B1 (V152), Phospho-Cyclin B1 (Ser133) (9E3), p53 (1C12) and HA-Tag (C29F4), Cell Signaling Technology, Danvers, MA). Horse radish peroxidase conjugated secondary antibodies were purchased from GE Healthcare Life Sciences (Pittsburgh, PA), and IRDye conjugated secondary antibodies (IRDye®800CW, 926-32210 and IRDye®680RD, 926-68071) were purchased from LI-COR Biotechnologies (Lincoln, NE).
Cyclin b1 v152
Manufactured by Cell Signaling Technology
Sourced in United States
Cyclin B1 (V152) is a monoclonal antibody that recognizes the Cyclin B1 protein. Cyclin B1 is a key regulator of the cell cycle, playing a crucial role in the progression from the G2 phase to mitosis.
Automatically generated - may contain errors
Lab products found in correlation
Horseradish peroxidase conjugated secondary antibody, by GE Healthcare (1 mentions)
Phospho histone h2a x ser139, by Cell Signaling Technology (1 mentions)
Ac 15, by Merck Group (1 mentions)
Cleaved caspase 3 asp175, by Cell Signaling Technology (1 mentions)
Ha tag c29f4, by Cell Signaling Technology (1 mentions)
Doxorubicin, by Merck Group (1 mentions)
P70s6k, by Cell Signaling Technology (1 mentions)
Puromycin, by Cayman Chemical (1 mentions)
Gapdh kc 5g4, by Kangchen Biotech (1 mentions)
Fbxo22 13606 1 ap, by Proteintech (1 mentions)
Cdk4 sc 260, by Santa Cruz Biotechnology (1 mentions)
Phospho p70s6k, by Cell Signaling Technology (1 mentions)
Mg132, by MedChemExpress (1 mentions)
Lc3 2, by Cell Signaling Technology (1 mentions)
Vimentin r28, by Cell Signaling Technology (1 mentions)
Caspase 8 1c12, by Cell Signaling Technology (1 mentions)
3 protocols using cyclin b1 v152
1
Immunoblotting Procedure for Protein Analysis
2
Protein Quantification and Analysis
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Puromycin and G418 were purchased from Cayman Chemical Company (Ann Arbor, MI, USA). NEM and doxorubicin was purchased from Sigma-Aldrich Corporation (St. Louis, MO, USA). CHX and Mg132 were obtained from MedChemExpress (Shanghai, China). The following antibodies were used for either western blotting or immunohistochemical analysis: FBXO22 (13606–1-AP, Protein-tech, Wuhan, China), CDK2 (SC-748, Santa Cruz Biotechnology, CA, USA), CDK4 (SC-260, Santa Cruz Biotechnology, CA, USA), cyclinE1 (EP435E, Abcam, Cambridge, MA, USA), and GAPDH (KC-5G4; Kang Chen Bio-tech, Inc., Shanghai, China). The following antibodies were purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA): Akt (C67E7), phospho-Akt (Ser473) (D9E), Erk (137F5), p38 (D13E1), phospho-p38 (3D7), p70s6k (49D7), phospho-p70s6k (1A5), cyclin B1 (V152), cyclin D1 (92G2) and p21(12D1).
3
Western Blot Analysis of Cell Signaling
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The cells were treated with vehicle or KB2 for 24 h and 48 h. The cells were lysed in whole-cell lysis buffer and western blotting was carried out as previously described (Bleloch et al., 2019 (link)). The primary antibodies used in this study were, rabbit polyclonal antibodies to phospho-histone H2A.X (Ser139) (#2577), cleaved caspase-3 (Asp175) (#9661), PARP (#9542), caspase-9 (#9502), LC3II (#2775), rabbit monoclonal antibody to β-Catenin (D10A8) (#8480), cleaved caspase-7 (Asp198) (D6H1) (#8438), mouse monoclonal antibodies to E-cadherin (4A2) (#14472), vimentin (R28) (#3932), Cyclin B1 (V152) (#4135), Caspase-8 (1C12) (#9746) from Cell Signaling Technology (Massachusetts, USA); mouse monoclonal antibody to p53 (DO-1) (sc-126), rabbit polyclonal antibodies to p21 (C-19) (sc-397), Cyclin A (H-432) (sc-751) from Santa Cruz Biotechnology (Texas, USA); rabbit polyclonal antibody to p38 MAP kinase (M0800) from Sigma Aldrich. After primary antibody incubation, membranes were incubated with goat anti-rabbit or goat anti-mouse HRP-conjugated secondary antibodies (Bio-Rad Laboratories, California, USA). p38 was used as a loading control. Densitometry readings were obtained using ImageJ and protein expression levels are represented as a ratio of protein of interest/p38 normalized to the vehicle control sample.
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