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Mouse anti actin

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Mouse anti-actin is a primary antibody that specifically recognizes the actin protein, a ubiquitous and essential component of the eukaryotic cytoskeleton. This antibody can be used to detect and quantify actin expression in various cell and tissue samples through techniques such as Western blotting, immunohistochemistry, and immunocytochemistry.

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Lab products found in correlation

Mouse anti flag, by Merck Group (4 mentions) Horseradish peroxidase hrp conjugated secondary antibody, by ZSGB-BIO (2 mentions) Rabbit anti akt, by Abcam (2 mentions) Rabbit anti p akt, by Abcam (2 mentions) Bca protein assay kit, by Merck Group (2 mentions) Rabbit anti caspase 3, by Abcam (2 mentions) Protein a g bead, by Santa Cruz Biotechnology (2 mentions) Anti flag magnetic beads, by Selleck Chemicals (2 mentions) Protease inhibitor, by Roche (2 mentions) Nitrocellulose membrane, by Bio-Rad (2 mentions) Image studio lite, by LI COR (2 mentions) Odyssey fluorescent western blotting system, by LI COR (2 mentions) Rabbit anti beclin1, by Abcam (1 mentions) Rabbit anti p62, by Abcam (1 mentions) Goat anti mif, by Abcam (1 mentions) Horseradish peroxidase conjugated secondary antibody, by ZSGB-BIO (1 mentions) Bicinchoninic acid protein assay kit, by Merck Group (1 mentions) Rabbit anti lc3b, by Abcam (1 mentions) Mouse anti p62, by Abcam (1 mentions) Rabbit anti apn, by Abcam (1 mentions)

Spelling variants of this product

Anti-actin (59 citations) Mouse anti-β-actin antibody (22 citations) Mouse anti-β-actin monoclonal antibody (19 citations) Monoclonal mouse anti-β-actin (10 citations) Mouse anti-β-actin (66009-1-Ig, (5 citations) Mouse anti-actin antibody (4 citations) Mouse anti-Beta Actin (4 citations) Anti-TM (2 citations) Mouse anti-β-actin monoclonal antibody (66009-1-Ig; (2 citations) Rabbit anti-mouse β-actin (2 citations)

16 protocols using mouse anti actin

1

Quantifying Autophagy and Apoptosis Markers

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To determine APN, LC-3, Beclin 1, Akt/P-Akt, P62, C-caspase-3/P-caspase-3 levels, proteins were extracted from the cells by suspension in radioimmunoprecipitation assay (RIPA) buffer. Samples were centrifuged at 12,000 rpm at 4°C for 30 min, and the supernatants were recovered for analysis. The protein concentrations were determined using the Bradford protein method and the bicinchoninic acid (BCA) protein assay kit (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany). Protein (40 µg) was electrophoresed on a pre-cast bis-Tris polyacrylamide gel (12%), and then transferred onto a polyvinylidene difluoride (PVDF) membrane. Membranes were blotted with rabbit anti-APN (1:1,000), rabbit anti-LC3B (1:500), rabbit anti-beclin 1 (1:1,000), rabbit anti-Akt (1:1,000), rabbit anti-p-Akt (1:1,000), rabbit anti-P62 (1:1,000), rabbit anti-Caspase-3 (1:1,000) (all from Abcam, Cambridge, MA, USA) and mouse anti-actin (1:1,000; ProteinTech Group, Inc., Chicago, IL, USA), followed by horseradish peroxidase (HRP)-conjugated secondary antibodies (1:5,000; ZsBio, Beijing, China). Immunoblots were visualized using enhanced chemiluminescence (LAS-4000).
Wang X., Liu Y., Liu W., Zhang Y., Guo F., Zhang L., Cui M., Liu S, & Wu R. (2018). Ubenimex, an APN inhibitor, could serve as an anti-tumor drug in RT112 and 5637 cells by operating in an Akt-associated manner. Molecular Medicine Reports, 17(3), 4531-4539.
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2

Immunoblotting Protein Detection Protocol

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The proteins were separated on denaturing polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to nitrocellulose membrane. The membrane was blocked and incubated overnight with primary antibodies: rabbit anti-PAR (Trevigen, 4336-BPC-100), mouse anti-PARP-1 (Fisher, BDB11039), mouse anti-Flag (Sigma, F3165), Goat anti-MIF (Abcam, ab36146), or mouse anti-actin (Proteintech, 66009-1-AP) at 4°C, followed by donkey anti-mouse, donkey anti-goat or goat anti-rabbit IgG conjugated to HRP for 1 h at room temperature. After washing, the immune complexes were detected by the SuperSignal West Pico Chemiluminescent Substrate (Pierce).
Ruan Z., Lu Q., Wang J.E., Zhou M., Liu S., Zhang H., Durvasula A., Wang Y., Wang Y., Luo W, & Wang Y. (2021). MIF promotes neurodegeneration and cell death via its nuclease activity following traumatic brain injury. Cellular and molecular life sciences : CMLS, 79(1), 39.
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3

Protein Expression Analysis in Cells

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To determine APN, LC3B, JNK/P-JNK, Akt/P-Akt, P62, and cleaved-caspase-3/pro-caspase-3 levels, proteins were extracted from the cells by suspension in radioimmunoprecipitation assay buffer. Samples were centrifuged at 12,000 rpm at 4°C for 30 min, and the supernatants were recovered for analysis. The protein concentrations were determined using the Bradford protein method and the bicinchoninic acid protein assay kit (Sigma, St Louis, MO, USA). Protein (40 µg) was electrophoresed on a pre-cast bis-Tris polyacrylamide gel (8%~12%) and then transferred onto a polyvinylidene difluoride membrane. Membranes were blotted with rabbit anti-APN (1:1,000), rabbit anti-LC3B (1:500), rabbit anti-JNK (1:1,000), rabbit anti-p-JNK (1:1,000), rabbit anti-Akt (1:1,000), rabbit anti-p-Akt (1:1,000), mouse anti-P62 (1:1,000), rabbit anti-Caspase-3 (1:1,000) (all from Abcam, San Francisco, CA, USA), and mouse anti-actin (1:1,000; Proteintech, NY, USA), followed by horseradish peroxidase-conjugated secondary antibodies (1:5,000; ZsBio, Beijing, China). Immunoblots were visualized using enhanced chemiluminescence (LAS-4000).
Wang X., Liu Y., Wu R., Guo F., Zhang L., Cui M., Wu X., Zhang Y, & Liu W. (2018). Role of ubenimex as an anticancer drug and its synergistic effect with Akt inhibitor in human A375 and A2058 cells. OncoTargets and therapy, 11, 943-953.
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4

Autophagy Protein Immunodetection Assays

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The following antibodies were used for immunostaining or immunoblotting assays: mouse anti-LC3 (MBL, M152-3), rabbit anti-LC3 (2775S; Cell Signaling Technology), rabbit anti-p62 (MBL, PM045), rabbit anti-FIP200 (SAB4200135; Sigma-Aldrich), rabbit anti-FIP200 (17250-1-AP; Proteintech), rabbit anti-ULK1 (8054; Cell Signaling Technology), rabbit anti-Atg13 (13468S; Cell Signaling Technology), rabbit anti-ATL2 (16688-1-AP; Proteintech), rabbit anti-ATL3 (ab117819; Abcam), mouse anti-WIPI2 (ab105459; Abcam), mouse anti-VAPB (66191-1-IG; Proteintech), rabbit anti-VAPA (15275-1-AP; Proteintech), mouse anti-LAMP1 (553792; BD Biosciences), rabbit anti-PTPIP51 (20641-1-AP; Proteintech), rabbit anti-ATG101 (13492; Cell Signaling Technology), rabbit anti-Myc (2278S; Cell Signaling Technology), mouse anti-Myc (M5546; Sigma-Aldrich), mouse anti-GFP (11814460001; Roche), mouse anti-actin (60008-1-IG; Proteintech), and rabbit anti-HA (H6908; Sigma-Aldrich).
The following reagents were used in this study: wortmannin (PHZ1301; Life Technologies), bafilomycin (B1793; Sigma-Aldrich), rapamycin (R0395; Sigma-Aldrich), Lipofectamine 2000 (12566014; Life Technologies), Lipofectamine RNAi MAX (13778150; Life Technologies), and LysoTracker Deep Red (L12492; Thermo Fisher Scientific).
Liu N., Zhao H., Zhao Y.G., Hu J, & Zhang H. (2021). Atlastin 2/3 regulate ER targeting of the ULK1 complex to initiate autophagy. The Journal of Cell Biology, 220(7), e202012091.
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5

Protein Expression Analysis in Hippocampus

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Cell lysates and mouse brain tissues comprising the hippocampi were prepared as previously described [7 (link)]. The protein extractions were analyzed by standard western blotting procedures and bands were visualized by Chemilluminescence. The following primary antibodies were used: mouse anti-actin (1:50,000, Proteintech, 66009-1-Ig, Rosemont, IL), rabbit anti-BDNF (1:3000, Genetex, GTX134514, Irvine, CA), rabbit anti-proBDNF (1:1000, Thermo Fisher Scientific, PA5-77533, Carlsbad, CA), rabbit anti-TrkB (1:1000, Proteintech, 13129-1-AP), rabbit anti-CPE (1:1000, Proteintech, 13710-1-AP), and rabbit anti-FGF2 (1:1000, Bioss, bs-0217R, Beijing, China).
Jiang D., Liu H., Li T., Zhao S., Yang K., Yao F., Zhou B., Feng H., Wang S., Shen J., Tang J., Zhang Y.X., Wang Y., Guo C, & Tang T.S. (2024). Agomirs upregulating carboxypeptidase E expression rescue hippocampal neurogenesis and memory deficits in Alzheimer’s disease. Translational Neurodegeneration, 13, 24.
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6

Western Blot Analysis of DENV Infection

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U2OS cells were transfected with the indicated plasmids using XtremeGene360 (Roche). When specified, cells were infected with DENV for the indicated timepoints. Cells were lysed in 1x RIPA + protease inhibitor + 0.1% SDS cocktail (Sigma). Lysates were sonicated prior to being separated by SDS PAGE using a 4-20% Tris-glycine polyacrylamide pre-cast gel (BioRad) and transferred to nitrocellulose membranes. Following 30 min of blocking in PBS + 10% non-fat milk, membranes were probed with the indicated primary antibodies: mouse anti-V5 (Invitrogen), rabbit anti-GFP (Proteintech), mouse anti-GFP (Proteintech), mouse anti-actin (Proteintech, rabbit anti-actin (Proteintech), and rabbit anti-DENV NS3 (GeneTex) followed by near-infrared dye-conjugated secondary antibodies (LiCor) diluted in PBST + 5% non-fat milk and imaged on an Odyssey CLx imaging system (LiCor).
Corliss L., Holliday M, & Lennemann N.J. (2022). Dual-fluorescent reporter for live-cell imaging of the ER during DENV infection. Frontiers in Cellular and Infection Microbiology, 12, 1042735.
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7

Molecular Signaling Assay Protocol

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Whole cells were lysed with RIPA lysis buffer (Beyotime, China, Cat No.P0013B). After protein quantification with the BCA Protein Assay Kit (Thermo Fisher Scientific, CA, Cat No.23225), equal amounts of proteins were separated on SDS-PAGE, and transferred to a PVDF membrane. Antibodies against the following proteins were used: Rabbit-anti-NRF2, rabbit-anti-MLC2, rabbit-anti-phospho-MLC2 (Thr18/Ser19), rabbit-anti-FAK, rabbit-anti-phospho-FAK (Tyr397), and rabbit-anti-ERR1, all purchased from Cell Signaling (Cat No. 12721, 3672, 3674, 3285, 8556 and 13826, respectively). Mouse-anti-RhoA was purchased from Cytoskeleton (Cat No. ARH03, CA), and mouse-anti-actin was purchased from ProteinTech (Cat No. 60008-1-Ig, CA).
Zhang C., Wang H.J., Bao Q.C., Wang L., Guo T.K., Chen W.L., Xu L.L., Zhou H.S., Bian J.L., Yang Y.R., Sun H.P., Xu X.L, & You Q.D. (2016). NRF2 promotes breast cancer cell proliferation and metastasis by increasing RhoA/ROCK pathway signal transduction. Oncotarget, 7(45), 73593-73606.
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8

Quantification of Inflammasome Activation

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Primary antibodies were mouse anti-capase-1 (AG-20B-0042; Adipogen), mouse anti-FLAG (F1804; Sigma–Aldrich), mouse anti-actin (66009–1-Ig; ProteinTech), anti-ASC (67824; Cell Signaling Technology), anti-GFP (BM3883; Boster Biotechnology), and anti-GSDMD (209845; Abcam). Mouse IL-18 enzyme-linked immunosorbent assay (ELISA) kit was purchased from Invivogen (BMS618). An LDH Cytotoxicity kit was obtained from ThermoFisher (C20300).
Shen C., Li R., Negro R., Cheng J., Vora S., Fu T.M., Wang A., He K., Andreeva L., Gao P., Tian Z., Flavell R., Zhu S, & Wu H. (2021). Phase Separation Drives RNA Virus-Induced Activation of the NLRP6 Inflammasome. Cell, 184(23), 5759-5774.e20.
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9

Quantifying FASN Protein Levels

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To determine the FASN level, proteins were extracted from the tissues by suspension in radioimmunoprecipitation assay (RIPA) buffer (Solarbio). The samples were centrifuged at 13,523 × g at 4°C for 38 min, and the supernatants were recovered for analysis. The protein concentration was determined using the bicinchoninic acid (BCA) protein assay kit (Sigma-Aldrich; Merck KGaA). Protein samples (40 µg) were electrophoresed on a pre-cast bis-Tris polyacryl-amide gel (8-12%) and then transferred onto a polyvinylidene difluoride membrane (EMD Millipore). The membranes were blotted with rabbit anti-FASN (1:1,000) and mouse anti-actin (all from ProteinTech Group, Inc.) followed by horseradish peroxidase (HRP)-conjugated secondary antibodies (1:5,000; ZsBio). Immunoblots were visualized using enhanced chemiluminescence (LAS-4000; GE Healthcare).
Wang X., Du G., Wu Y., Zhang Y., Guo F., Liu W, & Wu R. (2019). Association between different levels of lipid metabolism-related enzymes and fatty acid synthase in Wilms’ tumor. International Journal of Oncology, 56(2), 568-580.
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10

Antibodies and Reagents for Immunological Assays

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Rabbit anti-DYKDDDDK tag (D6W5B), rabbit anti-RIG-I (D14G6), rabbit anti-IRF3 (D83B9), rabbit anti-pIRF3 (4D46), rabbit anti-TBK1 (3031S), rabbit anti-pTBK1 (D52C2), and rabbit anti-TRAF3 antibodies (Abs) were obtained from Cell Signaling Technology (USA); the mouse anti-MAVS (E-3) and mouse anti-TRAF3 (G-6) antibodies were obtained from Santa Cruz Biotechnology (USA); the mouse anti-actin and rabbit anti-calnexin antibodies were obtained from Proteintech (Wuhan, China); the mouse anti-Flag M2 antibody was obtained from Sigma-Aldrich (USA); the mouse anti-Myc (9E10) antibody was obtained from Origene (USA); the rabbit anti-GM130 antibody was obtained from Abcam (United Kingdom); the mouse anti-GAPDH antibody (AF0006) was obtained from Beyotime (China); and the mouse anti-HA antibody was obtained from MDL Biotech (China). Alexa Fluor 488 goat anti-rabbit IgG secondary antibody, Alexa Fluor 568 goat anti-mouse IgG secondary antibody, Alexa Fluor 488 goat anti-mouse IgG secondary antibody, and Alexa Fluor 568 goat anti-rabbit IgG secondary antibody were obtained from Thermo Fisher Scientific. Protein A/G beads were obtained from Santa Cruz Biotechnology, and anti-Flag magnetic beads were obtained from Bimake (USA). Recombinant proteins of human IFN-β (P5660) and human IFN-λ1 (P5669) were purchased from Beyotime (China).
Zheng Y., Zhuang M.W., Han L., Zhang J., Nan M.L., Zhan P., Kang D., Liu X., Gao C, & Wang P.H. (2020). Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) membrane (M) protein inhibits type I and III interferon production by targeting RIG-I/MDA-5 signaling. Signal Transduction and Targeted Therapy, 5, 299.
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