After treatments, cells or liver tissues were washed with sterile phosphate-buffered saline for 3 times and then subjected to total protein extraction by using an radio-immunoprecipitation assay (RIPA) kit from Sigma-Aldrich (Cat. No. R0278). For secreted protein, culture media were collected and centrifuged for RIPA extraction. Then protein samples were quantified with bicinchoninic acid assay (BCA) reagent from Bio-Rad (Cat. No. 5000001). Western blot analyses of all proteins were performed as described using β-actin as the internal control.
Ripa kit
Manufactured by Merck Group
Sourced in United States
The RIPA (Radioimmunoprecipitation Assay) Kit is a laboratory product designed to extract and solubilize proteins from cells and tissues for further analysis. It provides a standardized and consistent method for protein extraction and preparation.
Automatically generated - may contain errors
Lab products found in correlation
Bca reagent, by Bio-Rad (3 mentions)
Bca protein assay, by Thermo Fisher Scientific (2 mentions)
Ab181020, by Abcam (1 mentions)
Amersham imagequant 800, by Cytiva (1 mentions)
Mouse anti rabbit igg hrp, by Santa Cruz Biotechnology (1 mentions)
Mab374, by Merck Group (1 mentions)
P ampk, by Abcam (1 mentions)
Image lab statistical software, by Bio-Rad (1 mentions)
Gapdh, by Abcam (1 mentions)
5 protocols using ripa kit
1
Protein Extraction and Quantification
2
Western Blot Analysis of CXCR4 Expression
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Protein was isolated from endothelial cells with a RIPA Kit (Sigma Aldrich, R0278, Burlington, MA, United States) supplemented with protease and phosphatase inhibitors. Protein concentration was determined via BCA protein assay (Thermo Fisher Scientific, 23227, Oakwood, OH, United States) per manufacturer’s instructions. Protein lysates (40 μg/lane) were loaded for probing CXCR4 (1:500 dilution; Abcam, Ab181020, Waltham, MA, United States). Following primary antibody incubation (overnight at 4°C), blots were incubated (1 h at room temperature) with a mouse anti-rabbit IgG-HRP (1:3,000 dilution; Santa Cruz, SC-2357, Dallas, TX, United States). Immunoreactive bands were detected using a western blot imaging system (Cytiva, Amersham ImageQuant 800, Marlborough, MA, United States). GAPDH was used as a loading control (1:400 dilution; Millipore Sigma, MAB374, Burlington, MA, United States).
3
Protein Extraction and Quantification
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After treatments, cells or liver tissues were washed with sterile PBS for three times and then subjected to total protein extraction by using a RIPA kit from Sigma–Aldrich. For secreted protein, culture medium was collected and centrifuged for RIPA extraction. Then protein samples were quantified with BCA reagent from Bio-Rad. Western blot analyses of all proteins were performed as described using β-actin as the internal control.
4
Western Blot Analysis of Protein Expression
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Extracted proteins from cells and tissues as directed by the radioimmunoprecipitation assay (RIPA) kit (Sigma-Aldrich, USA). Added 5–10 μL of the collected protein samples to the SDS-PAGE gel sample holes. Primary antibodies against the following targets were used: SIRT1 (Cat.No. 9475, CST), NAMPT (Cat.No. 236874, Abcam), IDO (Cat.No. 13268-1-AP, Sanying), AMPK (Cat.No. 32047, Abcam), p-AMPK (Cat.No. 131357, Abcam) and GAPDH (Cat.No. 8245, Abcam). The intensity of the bands on the western blots was evaluated by Image Lab statistical software (Bio-Rad, USA).
5
Protein extraction and analysis
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After treatments, cells or liver tissues were washed with sterile phosphate buffer saline (PBS) for three times and then subjected to total protein extraction by using a RIPA kit from Sigma–Aldrich. For secreted protein, culture medium was collected and centrifuged for RIPA extraction. Then protein samples were quantified with BCA reagent from Bio-Rad. Western blot analyses of all proteins were performed as described using β-actin as the internal control.
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