Western blotting was performed with a SDS‐PAGE Electrophoresis System according to previous report 6 with the following antibodies: OLFM4 (Billerica, MA), MMP 9 (Cell Signalling Technology, Beverly, MA) and β‐actin antibody (CWBIO, Jiangsu, China).
β actin antibody
Manufactured by CWBIO
Sourced in China, United States
The β-actin antibody is a primary antibody that specifically binds to the β-actin protein, a highly conserved cytoskeletal protein found in eukaryotic cells. This antibody can be used to detect and quantify the presence of β-actin in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Quantity one software, by Bio-Rad (2 mentions)
Mmp 9, by Cell Signaling Technology (1 mentions)
Caveolin 1, by Abcam (1 mentions)
Myd88, by Abcam (1 mentions)
Flotillin 1, by Abcam (1 mentions)
Apc cy7 anti human cd3 antibody, by BioLegend (1 mentions)
Bca protein assay kit, by Bio-Rad (1 mentions)
Nf κb antibody, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Clathrin, by Abcam (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Apc anti human cd19 antibody, by BioLegend (1 mentions)
Trip6 antibody, by Abcam (1 mentions)
Ripa protein extraction reagent, by Beyotime (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Cell lysis buffer, by Beyotime (1 mentions)
Cxcl8, by Abcam (1 mentions)
Homologous secondary antibodies, by CWBIO (1 mentions)
Ecl chemiluminescence kit, by CWBIO (1 mentions)
Acetylcholine, by Merck Group (1 mentions)
5 protocols using β actin antibody
1
Western Blot Analysis of OLFM4 and MMP9
2
Antibody Detection in Immune Cells
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Antibodies to TLR7, caveolin-1, flotillin-1, MyD88, clathrin, and Ki-67 were purchased from Abcam, Inc. (Cambridge, MA, USA). NFκB antibody was purchased from Cell Signaling Technology, Inc. (Boston, MA, USA). β-actin antibody, HRP-conjugated anti-rabbit, and anti-mouse IgG were purchased from CWBio, Inc. (Beijing, CN). Mouse IgG2A anti-Human TLR7 PE-conjugated monoclonal antibody and mouse IgG2A PE-conjugated isotype control were purchased from R&D Systems, Inc. (Minneapolis, MN, USA). BV510 anti-human CD14 antibody and BV421 anti-human CD16 antibody were purchased from BD Biosciences (San Jose, CA, USA). APC/Cy7 anti-human CD3 antibody, PerCP/Cy5.5 anti-human CD8a antibody, FITC anti-human CD56 antibody, PE/Cy7 anti-human CD11c antibody and APC anti-human CD19 antibody were purchased from Biolegend (San Diego, CA, USA). Protease inhibitor cocktails were obtained from Roche Molecular Biochemicals (Mannheim, DE). BCA protein assay kit and PVDF membranes were purchased from Bio-Rad (Hercules, CA). MβCD, media (Dulbecco's modified Eagle's medium, DMEM), MTT and other chemicals were purchased from Sigma (St. Louis, MO, USA). Gardiquimod was purchased from Santa Cruz (California, USA). aPPD was provided by Shanghai Innovative Research Centre of Traditional Chinese Medicine (Shanghai, China). The compound was 97.9% pure as measured by HPLC analysis.
3
Western Blot Analysis of TRIP6 Protein
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Total protein was extracted from cells using RIPA protein extraction reagent (Beyotime, China) and was quantified using the BCA protein assay kit (Beyotime, China). Equal amounts (50 μg) of protein was separated by 10% SDS-PAGE and transferred on PVDF membrane. The target proteins were incubated with the following primary antibodies: TRIP6 antibody (dilution: 1:1000, Abcam, U.S.A.) or β-actin antibody (dilution: 1:3000, CWBIO, China). Signals were detected using enhanced chemiluminescence reagents (Pierce, U.S.A.) and analyzed by Quantity One Software (Bio-Rad, U.S.A.).
4
Protein Extraction and Western Blot
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Total protein was extracted using cell lysis buffer (Beyotime) for Western blot. Equal amounts of protein were denatured and then separated by SDS/PAGE (12% gel). The target proteins were incubated with the following primary antibodies: CXCL8 (Abcam) or β-actin antibody (CWBio). Then the proteins were incubated with homologous secondary antibodies (CWBio). For HRP detection, an ECL chemiluminescence kit (CWBio) was used. Intensity of blots was performed by Quantity One Software (Bio–Rad).
5
Pharmacological Evaluation of SGD Herbal Formula
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White attractylodes rhizome (Baizhu) (Shang Hai De Hua GuoYao; Lot number: 2018061101), white peony root (Baishao) (Shang Hai Hua Pu Zhong Yao; Lot number: 2018042901), dried old orange peel (Chenpi) (Shang Hai Lei Yun Shang Zhong Yao; Lot number: 1805037), ledebouriella root (Fangfeng) (Shang Hai Yu Tian Cheng Zhong Yao; Lot number:2017022706), and Radix bupleuri (Chaihu) (Ma Chen Jiu Zhou; Lot number: E2018050101), which form the components of SGD, were purchased as crude herbs from the JinKe Pharmacy (Shanghai, China). Saikosaponin A (National Institute for Food and Drug Control; Lot number: 110777-201912), paeoniflorin (National Institute for Food and Drug Control; Lot number: 110736-201943), 5-O-Methylvisammioside (National Institute for Food and Drug Control; Lot number: 111523-201811), hesperidin (National Institute for Food and Drug Control; Lot number: 110721-201818), and cimicifugoside (National Institute for Food and Drug Control; Lot number: 111522-201913) were purchased from the Shanghai Zhaorui Biological Technology Co. (Shanghai, China). Muscarinic receptor subtype-2 (M2R) antibody (Abcam: ab109266); muscarinic receptor subtype-3 (M3R) antibody (Abcam: ab87199); HRP secondary antibody (CWBIO: 111-035-003); β actin antibody (CWBIO: ET1702-67); and acetylcholine (Sigma: C4382-10G) were additionally obtained.
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White attractylodes rhizome (Baizhu) (Shang Hai De Hua GuoYao; Lot number: 2018061101), white peony root (Baishao) (Shang Hai Hua Pu Zhong Yao; Lot number: 2018042901), dried old orange peel (Chenpi) (Shang Hai Lei Yun Shang Zhong Yao; Lot number: 1805037), ledebouriella root (Fangfeng) (Shang Hai Yu Tian Cheng Zhong Yao; Lot number:2017022706), and Radix bupleuri (Chaihu) (Ma Chen Jiu Zhou; Lot number: E2018050101), which form the components of SGD, were purchased as crude herbs from the JinKe Pharmacy (Shanghai, China). Saikosaponin A (National Institute for Food and Drug Control; Lot number: 110777-201912), paeoniflorin (National Institute for Food and Drug Control; Lot number: 110736-201943), 5-O-Methylvisammioside (National Institute for Food and Drug Control; Lot number: 111523-201811), hesperidin (National Institute for Food and Drug Control; Lot number: 110721-201818), and cimicifugoside (National Institute for Food and Drug Control; Lot number: 111522-201913) were purchased from the Shanghai Zhaorui Biological Technology Co. (Shanghai, China). Muscarinic receptor subtype-2 (M2R) antibody (Abcam: ab109266); muscarinic receptor subtype-3 (M3R) antibody (Abcam: ab87199); HRP secondary antibody (CWBIO: 111-035-003); β actin antibody (CWBIO: ET1702-67); and acetylcholine (Sigma: C4382-10G) were additionally obtained.
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