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Rabbit monoclonal anti β actin

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Rabbit monoclonal anti-β-actin is a primary antibody that recognizes the β-actin protein. β-actin is a widely expressed cytoskeletal protein that is essential for various cellular processes. This antibody can be used to detect and quantify β-actin in a variety of sample types using techniques such as western blotting, immunohistochemistry, and immunocytochemistry.

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Lab products found in correlation

Pierce bca protein assay kit, by Thermo Fisher Scientific (4 mentions) Phosphatase inhibitor 2 and 3, by Merck Group (2 mentions) Protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions) Cell lytic mt lysis buffer, by Merck Group (2 mentions) Anti rabbit igg secondary antibodies conjugated with horseradish peroxidase, by Cell Signaling Technology (2 mentions) Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (2 mentions) Pvdf membrane, by Bio-Rad (2 mentions) Protease and phosphatase inhibitors, by Sangon (1 mentions) Rapid blocking buffer, by Ipsen (1 mentions) Bicinchoninic acid protein assay kit, by Thermo Fisher Scientific (1 mentions) Rabbit monoclonal anti β catenin, by Cell Signaling Technology (1 mentions) Rabbit monoclonal anti axin 1, by Cell Signaling Technology (1 mentions) Horseradish peroxidase conjugated secondary antibody, by Abcam (1 mentions) Rabbit polyclonal anti igfbp 5, by Abcam (1 mentions) Chemodoc xrs detection system, by Bio-Rad (1 mentions) Rabbit monoclonal anti stat3, by Cell Signaling Technology (1 mentions) Rabbit monoclonal anti mtor, by Cell Signaling Technology (1 mentions) Protease inhibitor cocktail, by Yeasen (1 mentions) Phosphatase inhibitor, by Yeasen (1 mentions) Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)

15 protocols using rabbit monoclonal anti β actin

1

Western Blot Analysis of Wnt/β-Catenin Pathway

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The HUVECs were seeded in a 6-well plate with a density of 2 × 105 cells per well. These cells were then treated with 1 μg/ml rhVEGF165 and combined with the previously indicated treatments. After 24 h of incubation, the cells were rinsed three times with PBS. The cell lysates were harvested with a lysis buffer with protease and phosphatase inhibitors (Sangon Biotech, China). After lysing on ice for 30 min, the cell lysates were centrifuged at 14,000 × g for 15 min at 4°C. The protein concentration was calculated using a bicinchoninic acid protein assay kit (Thermo Scientific, United States) and normalized. Thirty micrograms of protein were resolved with 6–10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electro-transferred to polyvinylidene fluoride membranes. The membranes were then blocked with a protein-free rapid blocking buffer (Epizyme, China) and immunoblotted with the following antibodies: rabbit monoclonal anti–β-catenin (Cell Signaling Technology, United States, 1:1000), rabbit monoclonal anti-Axin 1 (Cell Signaling Technology, United States, 1:1000), and rabbit monoclonal anti–β-actin (Cell Signaling Technology, United States, 1:1000). All quantitative data were normalized to β-actin as an endogenous control. ImageJ software was used to analyze the band intensity.
Zhong Y., Wang K., Zhang Y., Yin Q., Li S., Wang J., Zhang X., Han H, & Yao K. (2021). Ocular Wnt/β-Catenin Pathway Inhibitor XAV939-Loaded Liposomes for Treating Alkali-Burned Corneal Wound and Neovascularization. Frontiers in Bioengineering and Biotechnology, 9, 753879.
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2

Western Blot Analysis of mTOR Pathway

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Western blot analysis was performed as previously described 31 (link). Briefly, MDSCs were lysed in Cell Lytic MT lysis buffer (Sigma) with Protease Inhibitor Cocktail (Invitrogen) and phosphatase inhibitor 2 and 3 (Sigma) for 15 min on a shaker. After centrifugation for 20 min at 12 000×g (4°C), the supernatants were saved and protein concentrations of the samples were determined using the Pierce BCA Protein Assay Kit (Thermo Scientific). Equal amounts of protein (30 μg) were loaded onto SDS-polyacrylamide gels and blotted onto PVDF membranes (BioRad). Western blot analysis was performed using antibodies against mTOR, phospho-mTOR, p70S6K, phospho-p70S6K, S6, and phospho-S6 (rabbit monoclonal antibodies, 1: 1 000, Cell Signaling). Antibody against β-actin (rabbit monoclonal anti-β-actin, 1: 2 000, Cell Signaling) was used as a loading control. For detection, the membrane was incubated with anti-rabbit IgG secondary antibodies conjugated with horseradish peroxidase (1: 2 000, Cell Signaling). Bands were visualized using SuperSignal West Pico Chemiluminescent substrate (ThermoScientific Pierce).
Zhao T., Du H., Ding X., Walls K, & Yan C. (2014). Activation of mTOR Pathway in Myeloid-derived Suppressor Cells Stimulates Cancer Cell Proliferation and Metastasis in lal−/− Mice. Oncogene, 34(15), 1938-1948.
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3

Western Blot Analysis of mTOR Pathway

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Western blot analysis was performed as previously described 31 (link). Briefly, MDSCs were lysed in Cell Lytic MT lysis buffer (Sigma) with Protease Inhibitor Cocktail (Invitrogen) and phosphatase inhibitor 2 and 3 (Sigma) for 15 min on a shaker. After centrifugation for 20 min at 12 000×g (4°C), the supernatants were saved and protein concentrations of the samples were determined using the Pierce BCA Protein Assay Kit (Thermo Scientific). Equal amounts of protein (30 μg) were loaded onto SDS-polyacrylamide gels and blotted onto PVDF membranes (BioRad). Western blot analysis was performed using antibodies against mTOR, phospho-mTOR, p70S6K, phospho-p70S6K, S6, and phospho-S6 (rabbit monoclonal antibodies, 1: 1 000, Cell Signaling). Antibody against β-actin (rabbit monoclonal anti-β-actin, 1: 2 000, Cell Signaling) was used as a loading control. For detection, the membrane was incubated with anti-rabbit IgG secondary antibodies conjugated with horseradish peroxidase (1: 2 000, Cell Signaling). Bands were visualized using SuperSignal West Pico Chemiluminescent substrate (ThermoScientific Pierce).
Zhao T., Du H., Ding X., Walls K, & Yan C. (2014). Activation of mTOR Pathway in Myeloid-derived Suppressor Cells Stimulates Cancer Cell Proliferation and Metastasis in lal−/− Mice. Oncogene, 34(15), 1938-1948.
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4

Western Blot Analysis of Extracellular Matrix Proteins

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Proteins were extracted from whole cell lysates and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, then transferred to a polyvinylidene fluoride membrane. The following primary antibodies were used: rabbit polyclonal anti-IGFBP-5 (1:1500; Abcam, Cambridge, UK), rabbit polyclonal anti-proteoglycan (1:1200, Abcam), rabbit polyclonal anti-type I collagen (1:1500, Abcam), rabbit polyclonal anti-type V collagen (1:2000, Abcam), rabbit monoclonal anti-phosphorylated mTOR (p-mOTR, 1:1000, Cell Signaling Technology, Danvers, USA), rabbit monoclonal anti-mTOR (1:1000, Cell Signaling Technology), rabbit monoclonal anti-phosphorylated STAT3 (p-STAT3, 1:2000, Cell signaling Technology), rabbit monoclonal anti-STAT3 (1:1000, Cell Signaling Technology), and rabbit monoclonal anti-β-actin (1: 5000; Cell Signaling Technology). Membranes were then incubated with the horseradish peroxidase-conjugated secondary antibodies (1:4000; Abcam). The ECL western blot substrate kit was used to detect the respective bands (Abcam). The membranes were exposed using a ChemoDoc XRS detection system (Bio-Rad, Milan, Italy).
Pan J., Li K., Huang W, & Zhang X. (2017). MiR-137 inhibited cell proliferation and migration of vascular smooth muscle cells via targeting IGFBP-5 and modulating the mTOR/STAT3 signaling. PLoS ONE, 12(10), e0186245.
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5

Mitochondrial Dynamics Regulation in Cell Apoptosis

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Dulbecco’s Modified Eagle’s Medium (DMEM), penicillin-streptomycin solution, 0.25% trypsin/EDTA, and phosphate-buffered saline were purchased from Gibco. Fetal bovine serum (FBS) was purchased from Biological Industries. Rabbit monoclonal anti-Mfn1(Cat#: 14739S), rabbit monoclonal anti-Mfn2(Cat#: 9482S), rabbit monoclonal anti-Opa1(Cat#: 80471S), rabbit polyclonal anti-Drp1(Phospho Ser616 Cat#: 4494S), rabbit monoclonal anti-AMPKα (Phospho Thr172 Cat#: 2535S), and rabbit monoclonal anti-β-actin(Cat#: 4970T) were purchased from Cell Signaling Technology. Rabbit monoclonal anti-Fis1(Cat#: ab156865) and rabbit monoclonal anti-DRP1(Cat#: ab184247) were purchased from Abcam. DMSO, Mito-Tracker, BCA Protein Quantification Kit, RIPA lysis buffer, protease inhibitor cocktail, phosphatase inhibitors, SDS-PAGE Gel Preparation Kit, and other Western blotting reagents were purchased from Yeasen (Shanghai, China). Annexin V-Alexa Fluor 647/PI Apoptosis Detection Kit was purchased from Fcmacs (Nanjing, China). The mitochondrial membrane potential assay kit with JC-1 was purchased from Beyotime (Nanjing, China). Unless otherwise indicated, all other chemicals and reagents were purchased from Sigma-Aldrich.
Zheng Y., Luo A, & Liu X. (2021). The Imbalance of Mitochondrial Fusion/Fission Drives High-Glucose-Induced Vascular Injury. Biomolecules, 11(12), 1779.
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6

Investigating Endothelial Cell Barrier Regulation

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Rabbit polyclonal anti-ZO-1, mouse monoclonal anti-occludin, and mouse monoclonal anti-claudin5 were obtained from Thermo Fisher Scientific (Waltham, MA, USA). Rabbit monoclonal anti-β-actin, anti-phospho-NF-κB p65, and anti-NF-κB p65 were purchased from Cell Signaling Technology (Danvers, MA, USA). Goat polyclonal anti-Axl, anti-Mertk, and rat monoclonal anti-Tyro3 antibodies were obtained from R&D Systems (Minneapolis, MN, USA). Rabbit polyclonal anti-phospho-Axl, anti-phospho-Mertk, and anti-phospho-Tyro3 were purchased from Affinity (Cincinnati, OH, USA). Horseradish peroxidase (HRP)-conjugated and Alexa Fluor-conjugated secondary antibodies were purchased from Biosharp (Hefei, China). Axl small interfering RNA (siRNA) or nonspecific control siRNA with the recommended transfection reagents were all from RiboBio (Guangzhou, China). LPS (O55:B5) was purchased from Sigma-Aldrich (Oakville, Ontario, Canada). The In Vitro Vascular Permeability Assay kit was obtained from Merck Millipore (Etobicoke, Ontario, Canada). Recombinant Mouse Gas6 protein was from R&D Systems (Minneapolis, MN, USA). The chemiluminescence (ECL) Western blot detection kit was from Wenke Biotechnology (Zhejiang, China).
Ni J., Lin M., Jin Y., Li J., Guo Y., Zhou J., Hong G., Zhao G, & Lu Z. (2019). Gas6 Attenuates Sepsis-Induced Tight Junction Injury and Vascular Endothelial Hyperpermeability via the Axl/NF-κB Signaling Pathway. Frontiers in Pharmacology, 10, 662.
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7

Pregnenolone 16α-carbonitrile Metabolic Pathway

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Pregnenolone 16α-carbonitrile (PCN) with 98% purity (Cat# C3884) was purchased from ApexBio Technology (Houston, USA). Additionally, corn oil (Cat# C116025) and vancomycin (Cat# C10791097) were purchased from Aladdin Company (Shanghai, China).
Neomycin sulfate (Cat#N814740), metronidazole (Cat# M813526) and ampicillin (Cat# C116025) were obtained from Macklin (Shanghai, China). Rabbit polyclonal anti-CYP3A11 (Cat# A13484) antibody and anti-CYP2B10 (Cat# A1463) antibody were purchased from Abclonal (Wuhan, China). Rabbit monoclonal anti-β-actin (Cat# AC038) antibody was purchased from Cell Signaling Technology (Danvers, MA, USA). Rabbit polyclonal anti-ANKRD1 (Cat# D121628), anti-CYR61 (Cat# D322190) and anti-CTGF (Cat# D260212) were all provided by Sangon Biotechnology (Shanghai, China).
Wu T., Li L., Zhou W., Bi G., Jiang X., Guo M., Yang X., Fang J., Pang J., Fan S, & Bi H. (2024). Gut Microbiota Affects Mouse Pregnane X Receptor Agonist Pregnenolone 16α-Carbonitrile-Induced Hepatomegaly by Regulating Pregnane X Receptor and Yes-Associated Protein Activation. Drug metabolism and disposition: the biological fate of chemicals, 52(7).
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8

Antibody Characterization for Cellular Analyses

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The antibody used for western blot—rabbit monoclonal anti-β-actin—was purchased from Cell Signaling Technology, and mouse monoclonal anti-BAG1 was raised in our lab. For the immunofluorescence assay, mouse monoclonal anti-SAG1, goat anti-mouse IgG H&L (Alexa Fluor® 488), and goat anti-mouse IgG H&L (Alexa Fluor® 594) were purchased from Abcam (United Kingdom). The antibodies used for immunohistochemistry—anti-Ki67 rabbit polyclonal antibody, anti-CD3 rabbit polyclonal antibody, anti-CD4 rabbit polyclonal antibody, and anti-CD8 rabbit polyclonal antibody—were purchased from Servicebio (China).
Chen M., Yang P., Xin Z., Chen J., Zou W., Zhou L., Yang L., Peng J, & Peng H. (2023). Toxoplasma gondii gra5 deletion mutant protects hosts against Toxoplasma gondii infection and breast tumors. Frontiers in Immunology, 14, 1173379.
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9

Protein Extraction and Western Blot Analysis

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Protein extraction from macrophages was performed using ice-cold lysis buffer supplemented with protease & phosphatase inhibitor cocktail (Thermo Scientific). Equal amounts of protein lysates were resolved by SDS-PAGE. The primary antibodies applied in our study were as follows: rabbit monoclonal anti-NLRP3 (15,101 S, Cell Signaling Technology), rabbit polyclonal anti-IL1β (bs-6319R, Bioss), rabbit monoclonal anti-β-actin (4970T, Cell Signaling Technology). An ImageQuant LAS 4000 imaging system (GE Healthcare) was used for image acquisition and analyses.
Pan W., Zhou G., Hu M., Li G., Zhang M., Yang H., Li K., Li J., Liu T., Wang Y, & Jin J. (2024). Coenzyme Q10 mitigates macrophage mediated inflammation in heart following myocardial infarction via the NLRP3/IL1β pathway. BMC Cardiovascular Disorders, 24, 76.
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10

Immunoblotting and Immunofluorescence Analysis of Cellular Markers

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The following antibodies were used: mouse monoclonal anti-FLAG M2 (Sigma-Aldrich), mouse monoclonal anti-ubiquitinated protein (FK2; Nippon Bio-Test Laboratories), mouse monoclonal anti-Rab30 (Abcam, ab156774), rabbit monoclonal anti-Atg5 (Cell Signaling Technology, #4967), rabbit monoclonal anti-β-actin (Cell Signaling Technology, #4970), mouse monoclonal anti-LAMP1 (Santa Cruz, sc-20011), and rabbit monoclonal anti-GM130 (BD Transduction Laboratories, 610822). The secondary antibodies used for immunoblotting were horseradish peroxidase-conjugated anti-mouse or anti-rabbit IgG (Jackson Immunoresearch Laboratories). The fluorescent secondary antibodies used for immunofluorescence were Alexa Fluor 488-conjugated goat anti-mouse or anti-rabbit IgG, Alexa Fluor 594-conjugated goat anti-mouse or anti-rabbit IgG, or Alexa Fluor 660-conjugated goat anti-mouse or anti-rabbit IgG (Molecular Probes/Invitrogen).
Oda S., Nozawa T., Nozawa-Minowa A., Tanaka M., Aikawa C., Harada H, & Nakagawa I. (2016). Golgi-Resident GTPase Rab30 Promotes the Biogenesis of Pathogen-Containing Autophagosomes. PLoS ONE, 11(1), e0147061.
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