Ginkgo biloba L. leaf extract injection (GBE) was purchased from Youcare Pharmaceutical Group (Beijing, China, Approval No. H20070226, Supporting Information 1). Its initial concentration is 3.5 mg/ml and prepared in aqueous solvent with excipients such as sorbitol, ethanol, sodium hydroxide. Lipopolysaccharide (LPS, #L2880) were obtained from sigma, and prepared in DMEM medium at concentration of 10 mg/ml. Primary antibodies for COX-2 (#4842, 1:1000), IκB-α (L35A5) (#4814, 1:1000), p-IκB-α (14D4) (#2859, 1:1000), NF-κB p65 (D14E12) (#8242, 1:1000) and p-p65 (93H1) (#3033, 1:1000), NF-κB p50 (D4P4D) (#13586, 1:1000) were obtained from Cell Signaling Technology, Inc. (Danvers, MA, United States). Primary antibodies for TNF-α (7B8A11) (#60291-1-Ig, 1:1000), IL-6 (#21865-1-AP, 1:1000), GAPDH (1E6D9) (#60004-1-Ig, 1:1000), β-actin (2D4H5) (#66009-1-Ig, 1:1000) and Lamin B1 (#12987-1-AP, 1:1000) were obtained from Proteintech Group Inc. (Rosemount, IL, United States). Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco Laboratories.
Gapdh 1e6d9
Manufactured by Proteintech
Sourced in United States
GAPDH (1E6D9) is a monoclonal antibody that recognizes the GAPDH protein. GAPDH is a glycolytic enzyme involved in the conversion of glyceraldehyde 3-phosphate to 1,3-bisphosphoglycerate during the glycolysis pathway. This antibody can be used for the detection of GAPDH in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Cox 2, by Cell Signaling Technology (1 mentions)
Lipopolysaccharide lps l2880, by Merck Group (1 mentions)
Nf κb p65 d14e12, by Cell Signaling Technology (1 mentions)
P iκb α 14d4, by Cell Signaling Technology (1 mentions)
P p65 93h1, by Cell Signaling Technology (1 mentions)
β actin 2d4h5, by Proteintech (1 mentions)
Laminb1 12987 1 ap, by Proteintech (1 mentions)
Iκbα l35a5, by Cell Signaling Technology (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Protease inhibitor cocktail, by Nacalai Tesque (1 mentions)
Sample buffer, by Thermo Fisher Scientific (1 mentions)
β actin 13e5, by Cell Signaling Technology (1 mentions)
Protease phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
Lysis buffer, by Cell Signaling Technology (1 mentions)
β mercaptoethanol, by Merck Group (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
Facsverse flow cytometer, by BD (1 mentions)
Imagequant las 4000, by GE Healthcare (1 mentions)
4 protocols using gapdh 1e6d9
1
Ginkgo biloba Extract Inhibits Inflammation
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Ginkgo biloba L. leaf extract injection (GBE) was purchased from Youcare Pharmaceutical Group (Beijing, China, Approval No. H20070226, Supporting Information 1). Its initial concentration is 3.5 mg/ml and prepared in aqueous solvent with excipients such as sorbitol, ethanol, sodium hydroxide. Lipopolysaccharide (LPS, #L2880) were obtained from sigma, and prepared in DMEM medium at concentration of 10 mg/ml. Primary antibodies for COX-2 (#4842, 1:1000), IκB-α (L35A5) (#4814, 1:1000), p-IκB-α (14D4) (#2859, 1:1000), NF-κB p65 (D14E12) (#8242, 1:1000) and p-p65 (93H1) (#3033, 1:1000), NF-κB p50 (D4P4D) (#13586, 1:1000) were obtained from Cell Signaling Technology, Inc. (Danvers, MA, United States). Primary antibodies for TNF-α (7B8A11) (#60291-1-Ig, 1:1000), IL-6 (#21865-1-AP, 1:1000), GAPDH (1E6D9) (#60004-1-Ig, 1:1000), β-actin (2D4H5) (#66009-1-Ig, 1:1000) and Lamin B1 (#12987-1-AP, 1:1000) were obtained from Proteintech Group Inc. (Rosemount, IL, United States). Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco Laboratories.
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Ginkgo biloba L. leaf extract injection (GBE) was purchased from Youcare Pharmaceutical Group (Beijing, China, Approval No. H20070226, Supporting Information 1). Its initial concentration is 3.5 mg/ml and prepared in aqueous solvent with excipients such as sorbitol, ethanol, sodium hydroxide. Lipopolysaccharide (LPS, #L2880) were obtained from sigma, and prepared in DMEM medium at concentration of 10 mg/ml. Primary antibodies for COX-2 (#4842, 1:1000), IκB-α (L35A5) (#4814, 1:1000), p-IκB-α (14D4) (#2859, 1:1000), NF-κB p65 (D14E12) (#8242, 1:1000) and p-p65 (93H1) (#3033, 1:1000), NF-κB p50 (D4P4D) (#13586, 1:1000) were obtained from Cell Signaling Technology, Inc. (Danvers, MA, United States). Primary antibodies for TNF-α (7B8A11) (#60291-1-Ig, 1:1000), IL-6 (#21865-1-AP, 1:1000), GAPDH (1E6D9) (#60004-1-Ig, 1:1000), β-actin (2D4H5) (#66009-1-Ig, 1:1000) and Lamin B1 (#12987-1-AP, 1:1000) were obtained from Proteintech Group Inc. (Rosemount, IL, United States). Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco Laboratories.
2
Western Blotting of Liver Proteins
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For western blotting, a piece of liver was homogenized in PBS buffer with the protease inhibitor cocktail (Nacalai, Kyoto, Japan), and 50 µg of total protein was suspended in Laemmli SDS sample buffer. Antibodies are as follows: Anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1E6D9, Proteintech, Rosemont, Illinois, USA), anti-AMPK (ABV10739, ABGENT, San Diego, California, USA), anti-phospho AMPK (pT172) (40H9, Cell Signaling Technology, Danvers, Massachusetts, USA), anti-total Protein Kinase B (AKT) (200–401 N98, Rockland, Limerick, Pennsylvania, USA), and anti-phosho AKT (pS473) (D9E, Cell Signaling Technology).
3
Western Blot Analysis of EMT Markers
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Cells were collected and lysed in 1X lysis buffer (Cell Signaling) and 1X protease/phosphatase inhibitor cocktail (Thermo Scientific). Samples (30 μg protein) were boiled in sample buffer (Thermo Fisher Scientific) containing β-mercaptoethanol (Sigma-Aldrich), separated by SDS-PAGE electrophoresis in polyacrylamide gels, and transferred onto nitrocellulose membranes (Bio-Rad). Membranes were incubated overnight at 4 °C with the primary antibody (1:1000 dilution), and the secondary antibody for 1 hour (1:2000 dilution). Protein bands were detected with the ImageQuantLAS4000 digital imager. Antibodies against Vimentin (D21H3), Snail (C15D3), Slug (C19G7), p-NDRG1 (Thr346, D98G11), NDRG1 (D8G9), p-p65 (Ser536, 93H1), p65 (D14E12) and β-Actin (13E5) were from Cell Signaling; GSK3β, p-GSK3β (Tyr216) and p-GSK3β (Ser9) from Novus Biologicals; Twist (Twist2C1a) and MDR1 (D-11) from Santa Cruz; GAPDH (1E6D9) was from Proteintech.
4
Membrane Protein Detection in Nanoparticles
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The presence of membrane proteins on the surface of BNPs was determined by Western blotting as published [13 (link)]. Membrane-coated PLGA NPs were purified by centrifugation at 9600×g for 10 min. Whole T-cell-enriched culture extract, membrane fraction, and membranes isolated from coated PLGA NPs by centrifugation at 9,600×g for 10 min (30 µg protein) were subjected to electrophoresis and transferred onto a nitrocellulose membrane. Membranes were incubated overnight at 4 °C with primary antibodies against GAPDH (1E6D9, Proteintech) or Na+/K+-ATPase α1 (C464.6, Santa Cruz Biotechnology) (1:1,000 dilution), and the secondary antibody (Cell Signaling) (1:2,000 dilution) for 1 h at room temperature. The chemiluminescence signal was obtained with an ImageQuant LAS 4000 (GE Healthcare).
The surface expression of IC receptors on NExT was determined by flow cytometry (FACSVerse flow cytometer, BD Biosciences) with no threshold on the forward scatter to detect the nanoparticles as previously reported [6 (link), 36 (link)]. Briefly, NExT were incubated for 15 min at room temperature with anti-PD1, anti-LAG3, and anti-TIM3 antibodies, or their corresponding isotype, as described above. The results were analyzed by FlowJo.
The surface expression of IC receptors on NExT was determined by flow cytometry (FACSVerse flow cytometer, BD Biosciences) with no threshold on the forward scatter to detect the nanoparticles as previously reported [6 (link), 36 (link)]. Briefly, NExT were incubated for 15 min at room temperature with anti-PD1, anti-LAG3, and anti-TIM3 antibodies, or their corresponding isotype, as described above. The results were analyzed by FlowJo.
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