Ki67 Immunofluorescence on Bovine Cell Lines
Ki67 免疫荧光法检测牛细胞系
发布: 2013年11月05日第3卷第21期 DOI: 10.21769/BioProtoc.958 浏览次数: 10688
评审: Fanglian He
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参见作者原研究论文
The authors used this protocol in:
Apr 2013
Abstract
This is a rapid protocol to test the effects of drugs treatment on bovine cell replication using Ki67 staining. Ki67 is associated with cell proliferation and is present during all active phases of the cell cycle (G1, S, G2, and mitosis), but is absent from resting cells (G0).
Keywords: Ki67 (Ki67)
Immunofluorescence (免疫荧光)
Bovine lymphocytes (牛淋巴细胞)
Materials and Reagents
- Bovine cells (The TBL3 cell line was derived from in vitro infection of the spontaneous bovine-B lymphosarcoma cell line, BL3, with Hissar stock of T. annulata)
- RPMI 1640 (Gibco)
- Fetal calf serum (heat-inactivated)
- Penicillin/streptomycin
- Fibronectin (Sigma-Aldrich, catalog number: F1141 )
- Formaldehyde (Sigma-Aldrich, catalog number: F8775 )
- Triton X-100 (Sigma-Aldrich, catalog number: T9284 )
- SVF (PAA Laboratories GmbH, catalog number: A15-101 )
- BSA (Sigma-Aldrich, catalog number: A2153 )
- Mouse monoclonal anti-Ki67 (1:50) (Abcam, catalog number: ab10913-1 )
- Cy2 AffinyPure anti-mouse IgG (1:5,000) (Jackson ImmunoResearch Laboratories, catalog number: 715-225-150 )
- Tween 20 (Biosolve, catalog number: 2045 2335 )
- ProLong Gold Antifade Reagent with Dapi (Life Technologies, Invitrogen™, catalog number: P-36931 )
- DAPI
Equipment
- Slides (Knittel Glass, catalog number: KN00010025787 )
- 37 °C, 5% CO2 cell culture incubator
- 24 wells plate
- Fluorescent microscope (Leica Microsystems, model: Inverted 6000 )
Procedure
文章信息
版权信息
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
- Marsolier, J., Medjkane, S., Perichon, M. and Weitzman, J. B. (2013). Ki67 Immunofluorescence on Bovine Cell Lines. Bio-protocol 3(21): e958. DOI: 10.21769/BioProtoc.958.
- Marsolier, J., Pineau, S., Medjkane, S., Perichon, M., Yin, Q., Flemington, E., Weitzman, M. D. and Weitzman, J. B. (2013). OncomiR addiction is generated by a miR-155 feedback loop in Theileria-transformed leukocytes. PLoS Pathog 9(4): e1003222.
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分类
细胞生物学 > 细胞成像 > 荧光
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