Cultured cells were lysed in RIPA buffer containing protease inhibitors. Protein concentration was determined by Bradford Protein Assay (Bioworld, 20830000-1). Equal protein per well was loaded and resolved by SDS-PAGE. Gels were transferred onto PVDF membranes (Thermo Scientific, 88518) and immunoblotted using the following antibodies: anti-PIM1 (Cell Signaling Technology-CST, 2907), anti-PIM2 (CST, 4730), anti-PIM3 (CST, 4165), anti-phospho IRS-1 (Ser1101-CST, 2385), anti-phospho GSK3β (Ser9-CST, 9323), anti- GSK3β (CST, 12456), anti-GAPDH (CST, 2118), anti-vinculin (CST, 13901), anti-HA-TAG (CST, 3924), anti-cyclin D1 (CST, 2922), anti-β-catenin (Santa Cruz Technology, SC-7963), anti-Tip47 (Santa Cruz Technology, SC-390981), anti-PPARα (LifeSpan BioSciences, LS-B46), anti-βactin (BD Transduction Laboratories, 612656). Anti-rabbit (CST, 7074) and Sheep a-mouse (Prometheus, 84–843) HRP secondary antibodies were used for detection. Blots were imaged on a ChemiDoc (SynGene) using chemiluminescence detection with ECL western blotting substrate (Thermo Scientific, 34095).
Anti β actin
Manufactured by BD
Sourced in United States
Anti-β-actin is a laboratory reagent used for the detection and quantification of the β-actin protein, a ubiquitous cytoskeletal protein found in eukaryotic cells. It is commonly used as a loading control or reference standard in various cell biology and molecular biology techniques, such as Western blotting and immunocytochemistry, to normalize the expression of target proteins.
Automatically generated - may contain errors
Lab products found in correlation
7500 fast real time pcr system, by Thermo Fisher Scientific (4 mentions)
Trizol reagent, by Thermo Fisher Scientific (4 mentions)
Alexa fluor 488 donkey anti rabbit, by Thermo Fisher Scientific (2 mentions)
Irdye 800cw donkey anti mouse, by LI COR (2 mentions)
Alexa fluor 488 donkey anti mouse, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 594 donkey anti rabbit, by Thermo Fisher Scientific (2 mentions)
Irdye 680cw donkey anti rabbit, by LI COR (2 mentions)
Alexa fluor 594 donkey anti mouse, by Thermo Fisher Scientific (2 mentions)
Anti rhoa, by Cell Signaling Technology (2 mentions)
Anti cdc42, by Cell Signaling Technology (2 mentions)
Anti pkcε, by Cell Signaling Technology (2 mentions)
Anti e cadherin, by R&D Systems (2 mentions)
Anti pkcε, by Santa Cruz Biotechnology (2 mentions)
Anti phospho smad2 3, by Cell Signaling Technology (2 mentions)
Anti phospho mypt1, by Cell Signaling Technology (2 mentions)
Anti pkcδ, by Cell Signaling Technology (2 mentions)
Anti vimentin, by Cell Signaling Technology (2 mentions)
Anti vinculin, by Merck Group (2 mentions)
Anti e cadherin, by BD (2 mentions)
Anti cox2, by BD (2 mentions)
32 protocols using anti β actin
1
Western Blot Analysis of Signaling Proteins
2
SARS-CoV-2 Antibodies and Antiviral Compounds
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Antibodies . The following antibodies were used for western blotting: ACE2 Polyclonal Goat IgG (cat n° AF933, R&D systems), Cathepsin B Monoclonal Mouse IgG1 (cat n° MA5-32651, Thermo Fisher Scientific), Cathepsin L Monoclonal Mouse IgG1 (cat n° BMS166, Thermo Fisher Scientific), anti-clathrin (cat. n° 610500, BD transduction laboratories), anti-β-actin (cat. n° MA1-140, Invitrogen), HRP-labeled goat anti-mouse immunoglobulin (cat. n° P0447, Dako). The following antibodies were used for flow cytometry: rabbit monoclonal SARS-CoV-2 spike-specific antibody (Cat. n° 40592-R001, Sino Biological), rabbit polyclonal SARS-CoV-2 nucleocapsid-specific antibody (Cat. n° GTX135357, GeneTex), fluorescent-labeled Alexa Fluor 647 (AF647) goat anti-Rabbit IgG monoclonal antibody (Cat. n° 4414, Cell Signaling Technologies).
Compounds . Monophosphoramidate prodrug Remdesivir GS-5734 (Cat. n° 469411000, Acros Organics); Molnupiravir (MK-4482 or EIDD-2801), a nucleoside analogue prodrug (Cat. n° S8969, Bio-Connect); Nirmatrelvir (PF-07321332), an irreversible inhibitor of SARS-CoV-2 viral main protease Mpro (Cat. n° S9866, Bio-Connect); purified SARS-CoV-2 spike-specific monoclonal rabbit primary antibody R001 (Cat. n° 40592-R001, Sino Biological).
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3
Western Blotting Protocol for Protein Analysis
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Western blotting was performed as previously described [31 (link), 32 (link)]. Briefly, tissues were homogenized and lyzed in protein lysis buffer on ice for 30 minutes. The collected protein samples were separated by SDS-PAGE, transferred to PVDF membrane and then blotted with indicated antibodies (anti-NF-κB p65 antibody: Cell Signaling Technology #8242; anti-β-actin: BD Transduction Laboratories™ 612656).
4
Immunoblotting and Immunofluorescence Protocols for COMMD4 and Cell Markers
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The following primary antibodies were used; anti-COMMD4 (Abcam, ab115169), anti-COMMD4 (Abcam, ab219115), CK7 (Abcam, ab9021), anti-β-actin (BD Biosciences, 612656), PARP1 (Cell Signaling, 9532), α-tubulin (Sigma-Aldrich, T5168) and lamin A/C (Cell Signaling, 4777). The following secondary antibodies were used for immunoblotting; IRDye® 800CW Donkey anti-mouse, IRDye® 800CW Donkey anti-goat and IRDye® 680CW Donkey anti-rabbit (LI-COR, Inc.). The following secondary antibodies were used for immunofluorescence; Alexa Fluor® 488 donkey anti-rabbit (Life Technologies, A21206), Alexa Fluor® 488 donkey anti-mouse (Life Technologies, A21202), Alexa Fluor® 594 donkey anti-mouse (Life Technologies, A21203) and Alexa Fluor® 594 donkey anti-rabbit (Life Technologies, A21207).
5
Western Blot Analysis of Key Signaling Proteins
6
Western Blot Analysis of Key Signaling Proteins
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Western blots were carried out as previously described (35 (link)), using the following antibodies: anti-Rac1 clone 23A8 (catalog # 05-389, Upstate Biotechnology, Lake Placid, NY), anti-phospho-Erk1/2 (catalog # 9101S), anti-PKCδ (catalog # 2058S), anti-PKCε (catalog # 2683S), anti-phosphoSmad2/3 (catalog # 8828S), anti-phospho-MYPT1 (catalog # 5163S), anti-vimentin (catalog # 3390S), anti-Cdc42 (catalog # 2466S), anti-RhoA (catalog # 2117S, Cell Signaling Technology, Beverly, MA), anti-PKCε (catalog # sc-208, Santa Cruz Biotechnology, Dallas, TX), anti-vinculin (catalog # V9131, Sigma-Aldrich, St. Louis, MO), anti-β-actin (catalog # A5441, BD Biosciences, Franklin Lakes, NJ), and anti E-cadherin (catalog # MAB1838, RD Systems, Minneapolis, MN).
7
Western Blot Analysis of Stem Cell Markers
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Cell lysates were separated by 10% sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis and transferred onto polyvinylidene fluoride (PVDF) membranes. After blocking with 5% fat-free milk in Tris-buffered saline, the following antibodies were used for western blotting: anti-Sox2 (1∶500), anti-ALDH1 (BD Biosciences, 1∶500), anti-Bmi1 (1∶500), anti-Oct4 (1∶500), anti-Nanog (1∶500), anti-vimentin (1∶500), anti-snail (1∶500), anti-β-catenin (1∶500), anti-E-cadherin (1∶500), and anti-β-actin (1∶1000) overnight at 4°C. All antibodies were obtained from Santa Cruz Biotechnology unless otherwise specified. After washing, the bound antibodies were visualized using horseradish peroxidase-conjugated anti-goat, ant-rabbit, or anti-mouse IgG (Thermo Fisher Scientific Inc., New York, NY) and the Immobilon Western Chemiluminescent HRP Substrate (Millipore, Billerica, MA) and subsequently visualized on X-ray films [21] (link).
8
Immunoblotting for Cell Cycle Regulators
9
Antibody Characterization for DNA Damage Response
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The antibodies used were as follows: anti-Banf1 N-terminus (SAB1409950, Sigma-Aldrich, 1:1000 for WB and 1:500 for IF), anti-Banf1 C-terminus (PRS40170604, Sigma-Aldrich, 1:1000 for WB and 1:500 for IF), anti-Emerin (5430, Cell Signalling Technology, 1:500 for IF), anti-Flag M2 Antibody (F3165, Sigma-Aldrich, 1:1000 for WB and 1:300 for IF), anti-γ-Tubulin (T6557, Sigma-Aldrich, 1:2000 for WB), anti-H3 (4499, Cell Signaling Technology, 1:2000 for WB), anti-β-actin (612656, BD Biosciences, 1:2000 for WB), anti-P-ATM S1981 (5883, Cell Signaling Technology 1:1000), anti-Mre11 (HPA002691, Sigma-Aldrich, 1:1000 for WB), anti-MDC1 (PLA0016, Sigma-Aldrich, 1:1000 for IF), anti-γ-H2AX (ab26350, abcam, 1:1000 for WB; 9718, Cell Signalling Technology, 1:1000 for IF, ab81299, abcam for ChIP), anti-p-DNA-PK S2056 (Abcam, ab124918, 1:1000 wb), anti-DNA-PKcs (12311, Cell Signaling Technology 1:1000 for western blot and ab70205 1:200 for IF), anti-Ligase IV (ab193353, Abcam, 1:1000), anti-Ku70 (ab92450, Abcam, 1:1000) and anti-nucleolin (14574, Abcam, 1:1000). Fluorescent secondary antibodies used were: Donkey anti-Mouse 800 nm (LiCor; IRDye 800CW 926–32212, 1:5000 for WB), Donkey anti-Rabbit (LiCor; IRDye 680LT 926–28023, 1:5000 for WB) and Alexa Fluor 488 (Cat# A32766, Molecular Probes, 1:200 for IF) and 594 (Cat# A32754, Molecular Probes, 1:200 for IF).
10
Immunoblotting and Immunofluorescence Protocols
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The following primary antibodies were used: anti-COMMD1 (Abcam, ab224727), (Invitrogen, MA5-26010), anti-β-actin (BD Biosciences, 612656), anti-gamma H2AX (Abcam, ab26350), anti-p53 Serine 15 (Cell Signaling, 9284), anti-p53 clone D0–7 (Sigma-Aldrich, p8999), anti-Chk2 Threonine 68 (Cell Signaling, 2661), anti-Chk2 (Cell Signaling, 2662), anti-ATM Serine 1981 (Cell Signaling, 13050), anti-ATM (Cell Signaling, 2873), anti-H2AX (Cell Signaling, 7631) anti-MDC1 (Abcam, 11169). The following secondary antibodies from LI-COR, Inc, were used for immunoblotting; IRDye® 800CW Donkey anti-mouse (926-32212) and IRDye® 680CW Donkey anti-rabbit (926-68073). The following secondary antibodies from Life Technologies were used for immunofluorescence; Alexa Fluor® 594 donkey anti-rabbit (A21207), Alexa Fluor® 594 donkey anti-mouse (A21203), Alexa Fluor® 488 donkey anti-rabbit (A21206) and Alexa Fluor® 488 donkey anti-mouse (A21202).
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