Anti β actin a5441

Manufactured by Merck Group

Sourced in United States, United Kingdom, Germany

Anti-β-actin (A5441) is a laboratory reagent used in various applications that require the detection and quantification of β-actin, a commonly expressed cytoskeletal protein. This product is intended for research use only.

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Lab products found in correlation

Anti flag m2 f1804, by Merck Group (5 mentions) Anti hif 1α nb100 479, by Novus Biologicals (3 mentions) Hybond, by Cytiva (3 mentions) Protease inhibitor cocktail, by Merck Group (3 mentions) Anti gapdh sc 25778, by Santa Cruz Biotechnology (3 mentions) Hrp conjugated secondary antibody, by GE Healthcare (3 mentions) Complete protease inhibitor cocktail, by Roche (3 mentions) Anti shp2 610822, by BD (3 mentions) Anti pan akt 11e7, by Cell Signaling Technology (3 mentions) Anti flag f1804, by Merck Group (3 mentions) Anti rl2 ma1 072, by Thermo Fisher Scientific (3 mentions) Hiperfect transfection reagent, by Qiagen (2 mentions) Anti hif 2α nb100 122, by Novus Biologicals (2 mentions) Hybond c extra, by Cytiva (2 mentions) Anti bax 610982, by Merck Group (2 mentions) Anti trail r2 2019, by Merck Group (2 mentions) Anti p53 do 1, by Santa Cruz Biotechnology (2 mentions) Enhanced chemiluminescence system, by Merck Group (2 mentions) Anti rac1 05 389, by Merck Group (2 mentions) Peroxidase conjugated goat anti rabbit 111 035 045, by Jackson ImmunoResearch (2 mentions)

Close spelling variants of this product

β-actin (6886 citations) Anti-β-actin (3256 citations) Anti-actin (1282 citations) A5441 (398 citations) β-actin (A5441) (88 citations) Anti-β-actin antibody (A5441) (24 citations) Actin (A5441) (22 citations) Anti-actin (A5441) (16 citations) Mouse anti-β-actin antibody (A5441) (6 citations) Anti-β-actin monoclonal antibody (A5441) (5 citations)

87 protocols using anti β actin a5441

Protein Expression Analysis via Western Blot

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Cells were treated according to different needs of the experiment and lysed with RIPA (Beyotime) on ice to collect cell proteins. The protein expression of the cells was then detected by western blot technology. The antibodies needed for the proteins detected in this experiment are: anti-RBCK1 (26367-1-AP, Proteintech), anti-HIF1α (SC-135151, Santa Cruz), anti-β-Actin (A5441, Sigma), anti-HA (MMS-101R, Biolegend), anti-Myc (60003-2-lg, Proteintech), and anti-Flag (20543-1-AP, Proteintech). After the protein was electrophoretic, transparabed, and blocked, we incubated the corresponding primary antibody and the secondary antibody of the primary antibody species. Finally, we visualized the fluorescent signal of the resulting protein using AI600 (GE), during which the membrane was pre-processed with an Immobilon Western Chemilum HRP Substrate Kit (Millipore Co, Billerica).

Niu Z., Fan J., Chen F., Yang H., Li X., Zhuang T., Guo C., Cao Q., Zhu J., Wang H, & Huang Q. (2022). RBCK1 regulates the progression of ER-positive breast cancer through the HIF1α signaling. Cell Death & Disease, 13(12), 1023.

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Western Blotting and ChIP Antibody Catalog

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Antibodies utilized in Western blotting and/or chromatin immunoprecipitation assay (ChIP) were purchased from the following companies: anti-heme oxygenase-1 (sc-7695), anti-Nrf2 (sc-13032X), anti-JNK (sc-571), normal rabbit IgG (sc-2027), normal mouse IgG (sc-2025), and anti-RNAPII (sc-899X) all from Santa Cruz Biotechnology; anti-lamin B (Ab-1) from Oncogene; anti-lactate dehydrogenase (AB1222) from Chemicon; phospho-specific antibodies of p38 MAPK (9211), MEK1/2 (9121), JNK (4688), ERK1/2 (9911), histone H3S10 (3377), and anti-Histone H3 (9715) all from Cell Signaling Technology; ChIP grade phospho-specific anti-H3S10 (ab14955), anti-RNAPIISer2 (ab5095), and anti-RNAPIISer5 (ab5131) from Abcam; and anti-β-actin (A5441) from Sigma.

Ray P.D., Huang B.W, & Tsuji Y. (2015). Coordinated Regulation of Nrf2 and Histone H3 Serine 10 Phosphorylation in Arsenite-activated Transcription of the Human Heme Oxygenase-1 Gene. Biochimica et biophysica acta, 1849(10), 1277-1288.

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Autophagy Pathway Protein Analysis

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The primary antibodies anti-LC3B (L7543), anti-SQSTM1 (P0067), anti-phosphor (p)-mTOR (SAB4504476), anti-mTOR (T2949), anti-PIK3C3 (V9764), and anti-β-actin (A5441) were provided by Sigma-Aldrich (St. Louis, MO, USA), while anti-ATG5 (D5F5U), anti-P70S6K (9202S), and anti-p-P70S6K (9206S) were provided by Cell Signaling Technology (Boston, MA, USA), and anti-LAMP1 (21997-1-AP) was provided by Proteintech (Chicago, IL, USA). Meanwhile, the TRITC-labeled goat anti-rabbit IgG (H + L) secondary antibody (ZF-0316) was provided by ZSGB-BIO (Beijing, China), whereas the IRDye^® 800CW Goat anti-Mouse IgG (H + L) (925-32210), IRDye^® 680RD Goat anti-Rabbit IgG (H + L) (926-68071), and IRDye^® 800CW Goat anti-Rabbit IgG (H + L) (925-32211) secondary antibodies were provided by Li-Cor Biotechnology (Lincoln, NE, USA).

Liu Z., Zhang L., Liu Y., Zhang H., Chen J., Feng G., Yang P., Sha F., Cui L, & Sun G. (2021). Identification of Compound CB-2 as a Novel Late-Stage Autophagy Inhibitor Exhibits Inhibitory Potency against A549 Cells. Life, 11(8), 865.

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Epigenetic Regulation by EGCG in Cells

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(–) Epigallocatechin-3-gallate (EGCG) and dimethyl sulfoxide (DMSO) were procured from Sigma (St Louis, MO). A 1000-fold concentrate of EGCG was prepared in DMSO and stored at −80 °C. Dulbecco’s Modified Eagle Medium (DMEM) and trypsin were purchased from Invitrogen (Carlsbad, CA). Mouse monoclonal antibody against human Bmi-1 (ab14389) and goat polyclonal antibody against Ring1B (ab3832) was purchased from Abcam Inc. (Cambridge, MA). Mouse monoclonal anti-Ezh2 (#612667) was obtained from BD Transduction Labs (San Jose, CA). Antibodies specific for histone H3 K27-trimethyl (H3K27me3) (07-449), and ubiquitinated lysine 119 of histone H2A (H2AK119ub) (AB10029) were from Millipore (Billerica, MA). Anti-rabbit procaspase 9 (9502), and anti-mouse procaspase 8 (9746) were purchased from Cell Signaling Technology (Danvers, MA). Mouse monoclonal anti-β-actin (A5441) was obtained from Sigma (St Louis, MO). Peroxidase-conjugated sheep anti-mouse IgG (NA931) and donkey anti-rabbit IgG (NA934) were purchased from GE Healthcare (Piscataway, NJ).

Balasubramanian S., Scharadin T.M., Han B., Xu W, & Eckert R.L. (2015). The Bmi-1 helix–turn and ring finger domains are required for Bmi-1 antagonism of (–) epigallocatechin-3-gallate suppression of skin cancer cell survival. Cellular signalling, 27(7), 1336-1344.

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JARID1D Epigenetic Regulation in Prostate Cells

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DU145 and PC3 prostate cell lines and HEK293T cells were purchased from American Type Culture Collection (ATCC), which confirms cell lines using short tandem repeat analysis, and were cultured within 15 times of passages. Cell culture reagents were obtained from Invitrogen/Gibco. Anti-JARID1D antibodies were from Bethyl (A310-624A) and Santa Cruz (sc-83944). Anti-JARID1A (A300-897A), anti-JARID1B (A301-813A), and anti-JARID1C (A301-034A) antibodies were from Bethyl. Anti-H3, anti-H3K4me3, and anti-H3K27me3 antibodies were from Millipore. Normal Rabbit IgG was from santa cruz (sc-2027). Anti-FLAG (M2, F1804) and anti-β–Actin (A5441) antibodies were from Sigma Aldrich. The anti-p84 antibody was from GeneTex. JARID1D and its catalytic mutant (HE/AA) were cloned into the pFlag-CMV2 vector (Sigma). We previously reported that JARID1D's catalytic mutant mJARID1D (HE/AA) in a baculoviral recombinant form is enzymatically inactive (7 (link)).

Li N., Dhar S.S., Chen T.Y., Kan P.Y., Wei Y., Kim J.H., Chan C.H., Lin H.K., Hung M.C, & Lee M.G. (2016). JARID1D is a suppressor and prognostic marker of prostate cancer invasion and metastasis. Cancer research, 76(4), 831-843.

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Western Blot Protein Analysis Protocol

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Protein samples (20 μg) were separated in premade NuPAGE Bis-Tris gels (Thermo Fisher) and transferred to polyvinylidene difluoride membranes. The following antibodies were used: anti-PDK1 ADI-KAP-PK112-D (Enzo Biochem, New York, NY), anti-PDK2 sc-100534 (Santa Cruz Biotechnology), anti-PDK3 serum (rabbit antiserum provided by Dr Robert. A Harris), anti-PDK4 ab214938 (Abcam, Cambridge, MA), anti-p-PDHE1α (Calbiochem, San Diego, CA), anti-HSP90 #4874 (Cell Signaling Technology, Danvers, MA), and anti-β-actin A5441 (Sigma-Aldrich). Proteins were visualized using an LAS-4000 (BD Biosciences) or iBright1500 (Thermo Fisher).

Lee H., Jeon J.H., Lee Y.J., Kim M.J., Kwon W.H., Chanda D., Thoudam T., Pagire H.S., Pagire S.H., Ahn J.H., Harris R.A., Kim E.S, & Lee I.K. (2022). Inhibition of Pyruvate Dehydrogenase Kinase 4 in CD4+ T Cells Ameliorates Intestinal Inflammation. Cellular and Molecular Gastroenterology and Hepatology, 15(2), 439-461.

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CFTR Expression in F508del HeLa Cells

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HeLa‐F508del cells were treated 24 h with VX‐809 (10 μM). Total protein fraction was extracted with RIPA buffer and Complete Mini EDTA‐free protease inhibitor cocktail (Roche Diagnostics, Indianapolis, IN). For Western blots, equal amounts of proteins were resolved by 7% SDS‐PAGE and transferred onto polyvinylidene fluoride membrane (GE Healthcare, LittleChalfont, UK). The membranes were probed with specific antibodies anti‐CFTR 24‐1 (1:200) (R&D Systems, Minneapolis, MN, USA) and anti‐β actin (A5441; 1/10,000) (Sigma‐Aldrich, St‐Louis, USA) and detected with horseradish peroxidase (HRP)‐conjugated goat anti‐mouse IgG (1:50,000) (Sigma–Aldrich, St‐Louis, USA). The signal was detected using the enhanced chemiluminescence ECL Advance kit (GE Healthcare) and visualized using the G:BOX‐iChemi (Syngene, SynopticsLTD, UK).

Rocca J., Manin S., Hulin A., Aissat A., Verbecq‐Morlot W., Prulière‐Escabasse V., Wohlhuter‐Haddad A., Epaud R., Fanen P, & Tarze A. (2016). New use for an old drug: COX‐independent anti‐inflammatory effects of sulindac in models of cystic fibrosis. British Journal of Pharmacology, 173(11), 1728-1741.

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Immunoblotting for HIF-1α and HIF-2α

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Enhanced chemiluminescence (ECL) reagents, nitrocellulose membranes (Hybond-C extra), and secondary Cy3-conjugated antibodies were from Amersham Pharmacia Biotech Inc. (Piscataway, NJ, USA). The following antibodies were used: anti-HIF-1α (NB100-479) and anti-HIF-2α (NB100-122) from Novus Biologicals; anti SB3 from Xeptagen; anti-total UbQ (sc-8017), anti-LaminA (sc-20680), anti-p53 (sc-6243), anti-p21 (sc-817), anti-gliceraldehyde-3-phosphate dehydrogenase (GAPDH) (sc-20357), and anti-ERK (sc-94) from Santa Cruz Biotechnology; anti-NEDD8 (ab81264) from Abcam; anti-NAE-1 (AP13067c) from Abgent; anti-UBE-1 (BML-PW8395-00251) from ENZO Life Sciences; anti-HO-1 (MA1-112) from Invitrogen; anti-Ub-48 (05-137) from Millipore; anti-P-ERK (#4696) from Cell Signaling Technology; anti-α-tubulin (T9026) and anti-β-actin (A5441) from Sigma Aldrich. DMOG (Dimethyloxalylglycine) and all other reagents of analytical grade were from Sigma Aldrich. HiPerfect Transfection Reagent was from Qiagen. Lactate Colorimetric/Fluorometric assay kit was from Biovision Incorporation.

Cannito S., Foglia B., Villano G., Turato C., C Delgado T., Morello E., Pin F., Novo E., Napione L., Quarta S., Ruvoletto M., Fasolato S., Zanus G., Colombatto S., Lopitz-Otsoa F., Fernández-Ramos D., Bussolino F., Sutti S., Albano E., Martínez-Chantar M.L., Pontisso P, & Parola M. (2019). SerpinB3 Differently Up-Regulates Hypoxia Inducible Factors-1α and -2α in Hepatocellular Carcinoma: Mechanisms Revealing Novel Potential Therapeutic Targets. Cancers, 11(12), 1933.

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Western Blot Analysis of Cell Signaling

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Cell lysates were prepared using complete lysis buffer (EMD Millipore, San Diego, CA), with protease and phosphatase inhibitor cocktails (Roche Diagnostics, Indianapolis, IN). Protein quantification was performed using DC protein assay from Bio-Rad (Hercules, CA). Western blot analysis was performed as described previously (Al-Azayzih et al. 2015 (link); Sabbineni et al. 2019 (link)). Antibodies used include Akt1 (2938), N-cadherin (4061), phosphorylated p38-MAPK (9211), total p38-MAPK (9212), phosphorylated Smad2/3 (8828), total Smad2/3 (8685), FoxC2 (12974), phosphorylated β-catenin (9561), total β-catenin (8480) and GAPDH (2118) from Cell Signaling Technology (Danvers, MA). Anti-β-actin (A5441) was from Sigma (St. Louis, MO), anti-eNOS (610297) was from BD Pharmingen (San Diego, CA), anti-ALK-5 (ab31013) was purchased from Abcam (Cambridge, UK) and anti-ALK-1 (NBP1-90254) was obtained from Novus Biologicals. Anti-phosphorylated-Smad1/5/8 (AB3848-I) from Millipore Sigma and anti-Smad1/Smad5/Smad8 (SAB2702532) from Sigma-Aldrich. Band densitometry was performed using NIH Image J software (Bethesda, MD).

Verma A., Artham S, & Somanath P.R. (2020). ALK-1 to ALK-5 ratio dictated by the Akt1-β-catenin pathway regulates TGFβ-induced endothelial-to-mesenchymal transition. Gene, 768, 145293.

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Apoptosis Pathway Protein Analysis

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NGEN (2,3‐dihydro‐5,7‐dihydroxy‐2‐[4‐hydroxyphenyl]‐4H‐1‐benzopyran‐4‐one), dithiothreitol (DTT), 4′,6‐diamindino‐2‐phenylinodole (DAPI), EGTA, leupeptin, and Triton X‐100 were bought from Sigma‐Aldrich Co. (St Louis, MO). Anti‐Bcl‐2 (sc‐7328), anti‐Bcl‐xL (sc‐7195), anti‐Bad (sc‐7869), and anti‐Bax (sc‐493) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Anti‐cytochrome c (Cat. No. 556432) was bought from PharMingen (San Diego, CA). Anti‐β‐Actin (A5441) was purchased from Sigma‐Aldrich Co. and anti‐PUMA (Cat. No. PC686) was bought from Oncogene Science, Inc. (Uniondale, NY). Anti‐cytochrome oxidase IV (Cat. No. 556423) was purchased from Molecular Probes (Eugene, OR). Secondary antibodies of peroxidase‐conjugated anti‐rabbit IgG (Cat. No. NA934), anti‐mouse (Cat. No. NA931) and the enhanced chemiluminescence (ECL) detection kits were obtained from Amersham Life Science (Buckinghamshire, UK). Inhibitors of Caspase‐3 (z‐DEVD‐fmk), caspase‐8 (z‐IETD‐fmk), and caspase‐9 (z‐LEHD‐fmk) were obtained from KAMIYA Biomedical Company (Seattle, WA). Reagents of caspase activity assay were purchased from R&D Systems (Minneapolis, MN). All other chemicals with analytical grade quality had been obtained through commercial sources.

Lu W., Yu C.R., Lien H., Sheu G, & Cherng S. (2020). Cytotoxicity of naringenin induces Bax‐mediated mitochondrial apoptosis in human lung adenocarcinoma A549 cells. Environmental Toxicology, 35(12), 1386-1394.

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