Proteins were detected using the following primary antibodies: anti-enterovirus VP1 clone 5-D8/1 antibody purchased from Dako (Denmark); Mouse anti-Ub, Mouse anti-MAT1, Mouse anti-CDK7, and Mouse anti-Cyclin H antibodies purchased from Santa Cruz (USA); Mouse monoclonal to RNA polymerase II CTD and Rabbit monoclonal to RNA polymerase II CTD (phosphor S5) antibodies purchased from Abcam (USA); Mouse anti β-actin purchased from Proteintech; Rabbit monoclonal CDK2 and CDK4, Rabbit monoclonal phosphor-CDK2 and CDK4, Mouse monoclonal pRb, Rabbit monoclonal pRb-phospho Ser780, pRb-phospho Ser795 and pRb-phospho Ser807/811 antibodies purchased from Cell Signaling (USA); Rabbit polyclonal Lamin B1, Mouse monoclonal c-Myc, Mouse monoclonal HA-Tag and Mouse Monoclonal Flag antibodies purchased from Sigma (USA); Goat anti-mouse IgG horseradish peroxidase-conjugated secondary antibody and goat anti-rabbit IgG horseradish peroxidase-conjugated secondary antibody purchased from Thermo Fisher Scientific (USA).
Mouse anti β actin
Manufactured by Proteintech
Sourced in United States, China, United Kingdom, Switzerland
The Mouse anti-β-actin is a primary antibody that specifically recognizes the β-actin protein. It is a useful tool for detecting and quantifying the expression of β-actin, a widely expressed cytoskeletal protein, in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (10 mentions)
Mouse anti gapdh, by Proteintech (5 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (4 mentions)
Mouse anti flag, by Merck Group (3 mentions)
Ripa lysis buffer, by Beyotime (3 mentions)
Rabbit anti e cadherin, by Cell Signaling Technology (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Protease inhibitor cocktail, by Thermo Fisher Scientific (3 mentions)
Anti rabbit ask1, by Abcam (3 mentions)
Rabbit anti p62, by Proteintech (3 mentions)
Bca kit, by Thermo Fisher Scientific (3 mentions)
Ag 20b 0042, by Adipogen (3 mentions)
Ag 20b 0014, by Adipogen (3 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Merck Group (2 mentions)
Goat anti mouse igg, by Proteintech (2 mentions)
Rabbit anti nlrp3, by Novus Biologicals (2 mentions)
Rabbit anti il 1β, by Abcam (2 mentions)
Rabbit anti tnf α, by Abcam (2 mentions)
Rabbit anti ev a71 vp1, by GeneTex (2 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (2 mentions)
84 protocols using mouse anti β actin
1
Comprehensive Protein Detection Methodology
2
Antibody Reagents for Western Blot
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The mouse anti-FLAG (Sigma-Aldrich, USA), mouse anti-Myc (Sigma-Aldrich, USA), rabbit anti-ATP6V1A (Sigma-Aldrich, USA), mouse anti-β-actin (Proteintech, China), mouse anti-HA epitope tag (Proteintech, China), mouse anti-GST (Sigma-Aldrich, USA), mouse anti-His epitope tag (Proteintech, China), rabbit IgG (Solarbio, China) and the secondary antibodies (HRP-conjugated anti-mouse IgG antibody [Sigma-Aldrich, USA], HRP-conjugated anti-rabbit IgG antibody [Genescript, China], FITC-conjugated goat anti-mouse IgG antibody [ZSGB. Bio, China], Alexa Fluor 647-labeled goat anti-rabbit IgG (H + L) [Beyotime, China]) and gold-labeled goat anti-mouse IgG antibody (Sigma-Aldrich, USA) were purchased from the indicated vendors. Murine sera against M, P, or G protein were produced in our laboratory.
3
Investigating Phosphatase Regulation of Alpha-Synuclein
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Both protease and phosphatase inhibitors were purchased from Roche (Basel, Switzerland). Primary antibodies used in western blotting are as follows: Mouse anti-β-actin (1:5000, Proteintech), Rabbit anti-p-α-syn (1:1000, Abcam), Mouse anti-p-α-syn (1:1000, Wako), Mouse anti-α-syn (1:1000, Santa Cruz), Rabbit anti-Human α-syn (1:2000, Abcam), Mouse anti-demethylate-PP2A (1:1000, Millipore), Mouse anti-PP2A (1:1000, BD biosciences), Mouse anti-β-tubulin (1:1000, Abcam), Mouse anti-PPP2R2A (1:1000, CST), Mouse anti-PPP2R2B (1:1000, Abcam), Rabbit anti-PPP2R2D (1:1000, Abcam), Mouse anti-PPP2R5A (1:500, Santa Cruz), Rabbit anti-PPP2R5D (1:5000, Abcam), Rabbit anti-PPP2R5E (1:1000, Abcam), Mouse anti-LCMT-1 (1:1000, Abcam), Rabbit anti-PME-1 (1:1000, Millipore), Mouse anti-GAPDH (1:3000, Sigma), Mouse anti-c-Myc (1:5000, Clontech), and Mouse anti-FLAG (1:500, Santa Cruz). Secondary antibodies are as follows: Goat anti-Rabbit IRdye680 (LI-COR Bioscience), Goat anti-Mouse IRdye680 (LI-COR Bioscience), Goat anti-Rabbit IRdye800 (LI-COR Bioscience), Goat anti-Mouse IRdye800 (LI-COR Bioscience), Goat anti-Rabbit Alexa Fluor 488 (Thermo Fisher Scientific). The inhibitor of protein phosphatase methylesterase-1 (PME-1), AMZ30, was purchased from MedChem Express (Shanghai, China). Dulbecco’s Modified Eagle’s Medium (DMEM), and fetal bovine serum (FBS) were purchased from Gibco.
4
Western Blot Analysis of Apoptosis Markers
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Western blotting was performed as described previously20 (link). Antibodies were listed as follow: rabbit anti-SLFN11 (Santa Cruz Biotechnology, CA, USA), rabbit anti-Bax, rabbit anti-Bcl2, rabbit anti-caspase3, rabbit anti-cleaved-caspase3 and mouse anti-β-actin (Proteintech, IL, USA).
5
Hepatitis B Virus Protein Expression Analysis
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Cells were lysed 72 h after transfection. Cell and tissue samples were lysed in RIRA (Sigma-Aldrich) buffer with PMSF (Sigma-Aldrich). Lysates were centrifuged for 10 min at 12000 × g, and protein in the supernatant was quantified using the BCA method (Pierce). SDS-PAGE and western blot analysis were performed according to standard procedures. Mouse anti-HBs, anti-HBx, anti-HBp, and anti-HBc antibodies were kindly provided by Dr. Quan Yuan from Xiamen University. Goat anti-MAN1A1 (Santa Cruz; 1:500 dilution), rabbit anti-MAN1A2 (Proteintech; 1:200 dilution), rabbit anti-MAN1B1 (GeneTex; 1:500 dilution), mouse anti-MAN1C1 (Abcam; 1:500 dilution) antibodies were used as the primary antibodies. Mouse anti-β-actin (Proteintech; 1:1000 dilution) was used as a reference for protein quantification. Goat anti-mouse IgG (Proteintech; 1:10000 dilution), goat anti-rabbit IgG (Proteintech; 1:10000 dilution), and donkey anti-goat IgG (Santa Cruz; 1:10000 dilution) were used as the secondary antibodies followed by enhanced chemiluminescence (ECL; ThermoFisher Scientific) detection.
6
Antibody Production and Characterization
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The antibody against B2 protein of RGNNV was produced by aBIOTECH company (Jinan, China) through immunizing rabbits with B2 protein expressed in E. coli. The affinity and specificity of anti-B2 serum were confirmed by positive band in wtRGNNV-infected SSN-1 cells, but no signal was detected in non-infected SSN-1 cells through Western blotting assay (Figure S1 ). Other antibodies and reagents used in the current study were mouse anti-β-actin (66009-1, ProteinTech Group, Chicago, IL, USA), horseradish peroxidase (HRP) goat anti-mouse IgG (AS003, Abclonal, Wuhan, China), HRP goat anti-rabbit IgG (AS014, Abclonal, Wuhan, China), Alexa Fluor® 488 goat anti-mouse IgG (A-11001, Invitrogen, Grand Island, NY, USA), 4′,6-diamidino-2-phenylindole (DAPI) (C1002, Beyotime, Shanghai, China), proteinase inhibitor cocktail (P8340, Sigma, Darmstadt, Germany), phosphatase inhibitor cocktail 3 (P0044, Sigma, Darmstadt, Germany).
7
Western Blot Analysis of Astrocyte Proteins
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Cultured astrocytes were lysed in RIPA buffer with 13 complete protease inhibitors (Roche). Protein levels were assessed with a Bradford assay with BSA as the standard. Approximately 10 µg of denatured proteins was separated by 8% SDS-polyacrylamide gel electrophoresis and blotted onto 0.22 µm PVDF membranes (Roche). Nonspecific binding was blocked with TBST (TBS-0.1% Tween-20) with 3% (w/v) nonfat milk for 2 h at room temperature. Membranes were incubated overnight at 4℃ in TBST with 5% milk and the following primary antibodies: rabbit anti-GFAP (1:1000, 20,044,021, Dako), rabbit anti-MAP2 (1:500, 17,490–1-AP, Proteintech), rabbit anti-S100 beta (1:1000, ab52642, Abcam), and mouse anti-β-actin (1:5000, 60,008–1-IG, Proteintech). Membranes were then incubated at room temperature for 2 h in TBST with 5% milk and secondary antibodies (1:5000, Invitrogen). Protein bands were detected by chemiluminescence (Tanon, Shanghai, China) and quantified by densitometry with ImageJ (ImageJ 7.0 software). Protein levels were normalized to the level of β-actin as a control.
8
Western Blot Analysis of Inflammasome Proteins
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Western blotting was routinely performed as previously reported [20 (link)]. Mouse anti-β-actin (1:10,000, Proteintech, Chicago, IL), rabbit anti-NLRP3 (1:500, Novus Biologicals, CO, USA), rabbit anti-caspase-1 (1:500, Proteintech), mouse anti-ASC (1:500, Santa Cruz, CA, USA), rabbit anti-GSDMD (1:1000, CST, Danvers, USA), mouse anti-pro-IL-1β (1:1000, Proteintech), rabbit anti-cleaved IL-1β (1:1000, Novus Biologicals), rabbit anti-pro-IL-18 (1:1000, Proteintech), rabbit anti-cleaved IL-18 (1:300, Bioss, Beijing, China), and rabbit anti-caspase-11 (1:1000, Novus Biologicals) were used. The densities of protein blots were quantified by using ImageJ software (NIH, Bethesda, MD) and normalized to the level of β-actin.
9
Western Blot Analysis of Inflammatory Markers
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The collected Raw264.7 cells and bone surrounding soft tissues were homogenized and lysed in lysis buffer including RIPA and phenylmethanesulfonyl fluoride (PMSF; Beyotime Institute of Biotechnology, Haimen, Jiangsu, China). The lysates (30 μg/lane) were separated on 10% gels and were transferred onto polyvinylidene difluoride membranes (Beyotime Institute of Biotechnology). The membranes were blocked with 5% (w/v) BSA in Tris-buffered saline with 0.1% (w/v) Tween 20 (TBST) and probed with primary antibodies: mouse anti-ABCA1 (1: 1000; Abcam Biotechnology), rabbit anti-ABCG1 (1: 2500; Abcam Biotechnology), rabbit anti-Cav-1 (1: 1500; Abcam Biotechnology), rabbit anti-TNF-α (1: 1000; Abcam Biotechnology), rabbit anti-IL-6 (1: 1000; Cell Signaling Technology), rabbit anti-IL-1β (1: 1,000; Abcam Biotechnology), and mouse anti-β-actin (1: 4000; Proteintech, Rosemont, USA) at 4°C overnight. After washing, the bound antibodies were detected using optimal horseradish peroxidase-conjugated secondary antibodies: goat anti-rabbit IgG (1: 10 000) and goat anti-mouse IgG (1: 10 000; Merck Millipore, Darmstadt, Germany) and were visualized using the enhanced chemiluminescence reagents (Bio-Rad Laboratories, Hercules, USA). The relative levels of the target proteins to that of control β-actin were analyzed densitometrically using the Quantity One software (version 4.0, Bio-Rad).
10
Investigating Autophagy Regulation via PHB2
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The following antibodies were used: rabbit anti-LC3B (Cell Signaling Technology, Boston, MA, USA, 3868), rabbit anti-PHB2 (Cell Signaling Technology, 14085), rabbit anti-SQSTM1/p62 (ABclonal Technology, Wuhan, China, A77580), normal rabbit IgG (ABclonal Technology, AC005), mouse anti-PHB2 (Proteintech, Wuhan, China, 66424-1-Ig), mouse anti-β-actin (Proteintech, 66009-1-Ig), rabbit anti-flag (Proteintech, 20543-1-AP), and rabbit anti-EV-A71 VP1 (Genetex, Irvine, CA, USA, GTX132339).
The following reagents were used: 3-methyladenine (3-MA; Selleck, Shanghai, China, S2767), rapamycin (Selleck, S1039), nontargeting siRNA (SiNC) and siRNA targeting PHB2 (SiPHB2) and ATP6AP2 (SiATP6AP2) (RiboBio, Guangzhou, China), Lipofectamine 3000 Reagent (Invitrogen, Carlsbad, CA, USA, L3000015), Lipofectamine RNAiMAX Reagent (Invitrogen, 13778-150), and primers for PCR and real-time quantitative PCR (RT-qPCR) (Invitrogen).
The following reagents were used: 3-methyladenine (3-MA; Selleck, Shanghai, China, S2767), rapamycin (Selleck, S1039), nontargeting siRNA (SiNC) and siRNA targeting PHB2 (SiPHB2) and ATP6AP2 (SiATP6AP2) (RiboBio, Guangzhou, China), Lipofectamine 3000 Reagent (Invitrogen, Carlsbad, CA, USA, L3000015), Lipofectamine RNAiMAX Reagent (Invitrogen, 13778-150), and primers for PCR and real-time quantitative PCR (RT-qPCR) (Invitrogen).
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