Cultured cells were fixed with 4% paraformaldehyde (Sigma) in PBS for 5 minutes at RT and permeabilized with 0.3% Triton X-100 (Sigma) for 20 minutes. Cells were then incubated with blocking solution consisting of PBS and 5% FBS at RT for 30 minutes. For immunofluorescence staining, primary antibodies against albumin (ALB), (rat, polyclonal, 1:50; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), α-fetoprotein (AFP) (mouse, mono, 1:50; Wuhan Boster Biological Technology, Ltd., Wuhan, China) and pan cytokeratin (panCK; mouse, mono, 1:50; Zhongshan Goldenbridge Biotechnology, Beijing, China) were used. Following incubation with the primary antibody, cells were incubated with FITC-labeled anti-rat, Dylight-conjugated anti-mouse or FITC-labeled anti-mouse (rabbit, polyclonal, 1:200; Jackson ImmunoResearch, West Grove, PA, USA) secondary antibodies for 1 h at 37°C and stained with DAPI (Sigma) to identify the cell nuclei. The cells were photographed with a structured illumination fluorescence microscope (DM IL LED; Leica Microsystems, Tokyo, Japan).
Panck
Manufactured by Zhongshan Golden Bridge Biotechnology
Sourced in China
PanCK is a laboratory equipment product developed by Zhongshan Golden Bridge Biotechnology. It is used for the detection and quantification of pan-cytokeratin, a group of proteins found in epithelial cells. The core function of PanCK is to provide a reliable and accurate method for the identification and analysis of various cell types.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Dm il led, by Leica Microsystems (1 mentions)
Radioimmunoprecipitation assay buffer, by Cell Signaling Technology (1 mentions)
Protease and phosphatase inhibitor, by Roche (1 mentions)
P mek ser217 221, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
β tubulin, by Cell Signaling Technology (1 mentions)
2 protocols using panck
1
Immunofluorescence Staining of Cultured Cells
2
Protein Expression Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells were lyzed with radioimmunoprecipitation assay buffer (Cell Signaling Technology) containing protease and phosphatase inhibitor (Roche, Indianapolis, IN). The antibodies were: b-tubulin, b-actin, FAK, P-FAK (Y397), ERK, P-ERK (Thr202/204), MEK, and P-MEK (Ser217/221) (Cell Signaling Technology), and Pan-CK (Zhongshan Golden Bridge Bio-technology, Beijing, China).
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