Total proteins from tobacco K326 inoculated leaves at 1 dpi and systematic leaves at 3, 5, 7, 9 dpi, as well as the upper two leaves of N. benthamiana plants were extracted using a Plant Protein Extraction Kit (Solarbio, Shanghai, China) and quantified using the BCA protein quantification kit (Beyotime, Shanghai, China), respectively. After the completion of electrophoresis, the proteins separated by 12% SDS-PAGE were transferred to 0.20 μm polyvinylidene fluoride (PVDF) membranes (Millipore, Billerica, USA), which were then incubated in blocking solution for one hour (Solarbio, Shanghai, China). PVY CP monoclonal antibody was used at dilution of 1: 1000 (Youlong, Shanghai, China). Beta-actin antibody was used at dilution of 1: 5000 (Proteintech, Chicago, USA). After incubation with primary and secondary antibodies (ABclonal, Wuhan, China), membranes were washed twice for 10 min with 1×TTBS, and then transferred into ECL solution (Millipore, Billerica, USA) to detect signals by Tanon Chemiluminescence Gel Imager (Tanon, Shanghai, China).
Beta actin antibody
Manufactured by Proteintech
Sourced in United States
The beta-actin antibody is a research tool used to detect the presence and quantify the levels of the beta-actin protein, which is a ubiquitous and highly conserved cytoskeletal protein found in all eukaryotic cells. The antibody can be used in various laboratory techniques, such as Western blotting, immunofluorescence, and immunohistochemistry, to study the expression and localization of beta-actin in different cell types and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Bca kit, by Thermo Fisher Scientific (3 mentions)
Blocking solution, by Solarbio (2 mentions)
Plant protein extraction kit, by Solarbio (2 mentions)
Ecl solution, by Merck Group (2 mentions)
Goat anti rabbit antibody, by Proteintech (2 mentions)
Goat anti mouse antibody, by Proteintech (2 mentions)
Protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions)
Bca protein quantification kit, by Beyotime (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Sybr mixture, by Takara Bio (1 mentions)
Pg lps, by Merck Group (1 mentions)
Real time pcr primer col 1, by GeneCopoeia (1 mentions)
Runx 2, by GeneCopoeia (1 mentions)
Il 18, by GeneCopoeia (1 mentions)
Il 1β, by GeneCopoeia (1 mentions)
Tnf α, by GeneCopoeia (1 mentions)
Gapdh, by GeneCopoeia (1 mentions)
Rt master mix, by Takara Bio (1 mentions)
Annexin 5 fitc pi cell apoptosis detection kit, by BestBio (1 mentions)
Pvdf membrane, by Sangon (1 mentions)
13 protocols using beta actin antibody
1
Quantifying Viral Protein Expression
2
Necroptosis Regulation in Bone Cells
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Pg-LPS was bought from Sigma (St. Louis, MO, USA). Rabbit RIP1, MLKL and beta-actin antibody was from Proteintech (USA), and mouse RIP3 antibody was from Santa (USA). Real-time PCR primer COL-1, OCN, RUNX-2, BSP, TNF-α, IL-1β, IL-18 and GAPDH were from Genecopoeia (USA), SYBR mixture was from Takara (Japan), RT Master Mix was from Takara (Japan). Inhibitor of Nec-1 (ab141053) was from Abcam (USA). Annexin V-FITC/PI: Cell apoptosis detection kit was from Bestbio Company (China). Nude mice were bought from Animal Center of the Fourth Military Medical University Animal Center.
3
Western Blot Analysis of Tobacco and N. benthamiana Proteins
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Total proteins of tobacco K326 and N. benthamiana were extracted using a Plant Protein Extraction Kit (Solarbio, Shanghai, China). The proteins separated by 12% SDS-PAGE electrophoresis and were transferred to 0.20 μm polyvinylidene fluoride (PVDF) membranes (Sangon Biotech, Shanghai, China), which were then incubated in blocking solution for 1 h (Solarbio, Shanghai, China). PVY CP antibody was used at a dilution of 1: 1000 (Youlong, Shanghai, China). Beta-actin antibody was used at a dilution of 1: 5000 (Proteintech, Chicago, USA). Secondary antibody was used at a dilution of 1: 10000 (ABclonal, Wuhan, China). After incubation with primary and secondary antibodies, membranes were washed twice for 15 min with 1 × TTBS. Finally, the membranes were transferred into ECL solution (Millipore, Billerica, USA) to detect signals by Tanon Chemiluminescence Gel Imager (Tanon, Shanghai, China).
4
Sodium Hydrosulfide and β2-Microglobulin Assay
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Sodium hydrosulfide (NaHS, a donor of H2S) and β2-microglobulin were purchased from Sigma (Sigma, St. Louis, MO, United States). Anti-p62 antibody was purchased from Cell signaling Technology (Boston, MA, United States). Beta-actin antibody was purchased from Proteintech (Danvers, MA, United States).
5
Neuroprotective Mechanisms of H2S and Trk Inhibition
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Formaldehyde (FA), Sodium hydrosulfide (NaHS, a donor of H2S), K252a (a selective pharmacological pan-Trk inhibitor) and nitro blue tetrazolium (NBT) were supplied by Sigma Chemical CO (St. Louis, MO, USA). Cell counting kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies, Inc. (Rockvile, MD, USA). Specific monoclonal antibody to BDNF was obtained from Epitomic Inc (Burlingame, UK). Specific monoclonal antibody to Bax was purchased from Abcam Technology (Cambridge, CB, UK) and Specific monoclonal antibody to Bcl-2 was purchased from Cell Signaling Technology, Inc (Beverly, MA, USA). Beta-actin antibody, Goat anti-mouse antibody, Goat anti-rabbit antibody, and Goat anti-Rat antibody were purchased from Proteintech (Danvers, MA, USA). Caspase-3, 4-hydroxy-2-trans-nonenal (4-HNE), and malondialdehyde (MDA) enzyme-linked immunosorbent assay (ELISA) Kits were bought from USCN Life Science Inc (Wuhan, Hubei, China). Bicinchoninic Acid (BCA) Protein Assay Kit was obtained from Beyotime Institute of Biotechnology (Shanghai, China)
6
Comprehensive Antibody Panel for Investigating Cell Signaling Pathways
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We utilized these antibodies for research: human Osteoglycin (OGN) antibody for Western blotting(WB)from R&D Systems, Vascular endothelial growth factor A (VEGF-A) antibody, hypoxia inducible factor-1α (HIF-1α) from Abcam; human and mouse Osteoglycin (OGN) for immunohistochemistry (IHC) from Sigma-Aldrich and Santa Cruz Biotechnology; Akt, phospho-Akt (Ser473), epidermal growth factor receptor (EGFR), phospho-epidermal growth factor receptor (EGFR, Y1068), Erk1/2 antibody, CD3, CD8, PTPRC, and FOXP3 from Cell Signaling Technology; beta-actin antibody from proteintech; AKT activator: sc79 from Selleck.
7
Protein Extraction and Western Blot Analysis
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Total cellular protein was extracted using RIPA lysis buffer and a protease inhibitor cocktail (Sigma-Aldrich). After centrifugation at 18,000×g for 10 min, the supernatant was collected and protein concentration was measured using a bicinchoninic acid assay (BCA) kit (Thermo Fisher Scientific). Equal amounts of proteins were subjected to SDS-PAGE and transferred onto PVDF membranes using a discontinuous (Tris/CAPS) buffer system (Bio-Rad Laboratories, Hercules, CA, USA). Western blot analyses were performed using the following antibodies: mouse anti p53 (aa20-25; #MCA1701, Bio-Rad), Anti-p53 antibody [PAb 240] (#ab26, Abcam, Cambridge, UK), Anti-c-Myc antibody [Y69]-ChIP Grade (#ab32072, Abcam), Beta Actin antibody (#CL594-60,008, proteintech, Tokyo, Japan), monoclonal anti-FLAG® M2 antibody (#F3165-1MG, Sigma-Aldrich), and anti-mouse IgG HRP-linked whole antibody sheep (#NA931-1ML, Streptavidin Sepharose High Performance; Cytiva, Tokyo, Japan), Goat Anti-Mouse IgG H&L (HRP) (#ab6789, Abcam), Goat Anti-Rabbit IgG H&L (HRP) (#ab6721, Abcam) according to the manufacturers’ protocols. Chemiluminescent signals were visualized using a ChemiDoc™ Touch instrument (Bio-Rad).
8
Ferroptosis-related Protein Regulation
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Ferric ammonium citrate (USA, 1185-57-5) was purchased from Sigma–Aldrich. Recombinant mouse IL-1β (Minneapolis, USA, # 401-ML) was purchased from R&D systems. Erastin (USA, S7242) and Ferrostatin-1 (USA, S7243) were purchased from Selleck. Reactive Oxygen Species Assay Kit (China, S0033) was purchased from Beyotime. C11 BODIPY Lipid Peroxidation Sensor (USA, D3861) was purchased from ThermoFisher. GPX4 (UK, ab125066, diluted 1:5000) antibody, P53 antibody (UK, ab131442, diluted 1:1000), SLC7A11 antibody (UK, ab175186, diluted 1:5000), ACSL4(UK, ab155282, diluted 1:10,000) antibody, Collagen II (UK, ab34712, diluted 1:200 for immunohistochemical staining) antibody, Collagen II (UK, ab185430, diluted 1:1000 for western blot) antibody, NQO-1 antibody (UK, ab80588, diluted 1:10,000), and Trx (UK, ab133524, diluted 1:10,000) antibody were purchased from Abcam. Nrf2 antibody (USA, 16396-1-AP, diluted 1:1000), HO-1 antibody (USA, 10701-1-AP, diluted 1:1000), Beta-actin antibody (USA, 66009-1-Ig, diluted 1:1000) and GAPDH antibody (USA, 60004-1-Ig, diluted 1:1000) were purchased from Proteintech.
9
H2S Modulation of Cell Stress Responses
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NaHS (a donor of H2S), Imipramine, dimethyl sulfoxide, specific monoclonal anti-CHOP antibody, and Sirtinol were purchased from Sigma. Imipramine was dissolved in phosphate buffered saline (PBS) as solution and was administered i.p. of 15 mg/kg. Specific monoclonal anti-GRP-78 antibody and anti-Sirt-1 antibody were purchased from Epitomics. Specific monoclonal anti-cleaved caspase-12 antibody was produced by Santa Cruz Biotechnology. Beta-actin antibody, goat anti-mouse antibody, and goat anti-rabbit antibody were purchased from Proteintech.
10
Quantification of Munc18-1 Expression
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The expression of Munc18-1 and the TMR mutant in neurons at DIV 13–14 was quantitated by western blotting using Munc18-1 antibody (mouse monoclonal, Proteintech #67,137–1-Ig) at a dilution of 1:5000 and Beta Actin antibody (mouse monoclonal, Proteintech #66,009–1-Ig) at a dilution of 1:20,000. The integrated densities of each band were analyzed by ImageJ (NIH).
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