Anti β actin 3700

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-β-actin (3700) is a primary antibody produced by Cell Signaling Technology. It is designed to detect β-actin, a ubiquitous cytoskeletal protein.

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Lab products found in correlation

Trypsin edta, by Thermo Fisher Scientific (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Dmem f12, by Thermo Fisher Scientific (2 mentions) Ab129002, by Abcam (1 mentions) Ab124892, by Abcam (1 mentions) Ab54481, by Abcam (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Loading buffer, by Thermo Fisher Scientific (1 mentions) Polyvinylidene difluoride membrane, by Merck Group (1 mentions) Immobilon western chemiluminescent hrp substrate, by Merck Group (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Non fat dry milk, by Bio-Rad (1 mentions) Ab15537, by Abcam (1 mentions) Ab179695, by Abcam (1 mentions) Ab121024, by Abcam (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Avantor (1 mentions) Tgf β3, by Thermo Fisher Scientific (1 mentions) Tgf β2, by Thermo Fisher Scientific (1 mentions) Ly2109761, by MedChemExpress (1 mentions) Repsox, by MedChemExpress (1 mentions)

15 protocols using anti β actin 3700

Protein Expression Analysis by Western Blot

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Cells were washed twice with ice‐cold PBS and ruptured with RIPA Lysis Buffer (#P0013B; Beyotime), PMSF, cocktail inhibitor and phosphatase inhibitor cocktail. Cell extracts were microcentrifuged for 15 minutes at 12 000 g and supernatants were collected. Cell lysates (15‐20 μL) were resolved by SDS‐PAGE and transferred onto PVDF membranes. Membranes were blocked for 1 hour with 5% non‐fat milk in TBST (Trisbuffered saline containing 0.1% Tween 20) and incubated overnight at 4°C with anti‐FTO antibody (ab124892; Abcam, Shanghai, China), anti‐β‐actin (3700S; Cell Signal Technology, Danvers, MA, USA), anti‐Vinculin (ab129002, Abcam), anti‐PGC‐1α (ab54481, Abcam). Membranes were washed 5 minutes with TBST for three times, incubated for 1 hour with required secondary antibodies conjugated to horseradish peroxidase and developed by chemiluminescent substrates.

Zhuang C., Zhuang C., Luo X., Huang X., Yao L., Li J., Li Y., Xiong T., Ye J., Zhang F, & Gui Y. (2019). N6‐methyladenosine demethylase FTO suppresses clear cell renal cell carcinoma through a novel FTO‐PGC‐1α signalling axis. Journal of Cellular and Molecular Medicine, 23(3), 2163-2173.

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Western Blot Analysis of CLL Cell Lysates

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CLL samples were processed for Western blotting analysis as previously described^{36 (link)}. Briefly, cells were lysed in RIPA buffer, containing 1% NP-40, 10 mM Tris-HCl, 150 mM NaCl, 0,5% Na-deoxycholate and 1 mM EDTA, supplemented with the protease inhibitor cocktail (complete-mini EDTA-free tablets—Roche) on ice for 30 min. Lysates were clarified by 30 min-centrifugation at 14,000xg, at 4 °C. Western blotting was performed by suspending protein aliquots in loading buffer (Thermo Fisher Scientific), resolved on Novex^® NuPAGE^® SDS-PAGE Precast Bis-Tris Gels 4 to 12%, transferred to PolyVinylidene-DiFluoride membrane (Millipore, Bedford, MA, USA), and incubated with primary antibodies followed by incubation with horseradish-peroxidase-linked mouse or rabbit IgG (1:2000) (GE Healthcare Amersham, Little Chalfont, UK) in PBS containing 5% non-fat dry milk (Bio-Rad Laboratories). Proteins were detected by chemiluminescence using the Immobilon Western Chemiluminescent HRP substrate (Millipore). Primary antibodies were purchased from Thermo Fisher Scientific (anti-IBtkα, PA5-24224) and Cell Signalling (anti- β-Actin, #3700S).

Albano F., Chiurazzi F., Mimmi S., Vecchio E., Pastore A., Cimmino C., Frieri C., Iaccino E., Pisano A., Golino G., Fiume G., Mallardo M., Scala G, & Quinto I. (2018). The expression of inhibitor of bruton’s tyrosine kinase gene is progressively up regulated in the clinical course of chronic lymphocytic leukaemia conferring resistance to apoptosis. Cell Death & Disease, 9(1), 13.

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TGF-β Isoforms, Hepatitis C Inhibitors

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TGF‐β1 (100‐21‐10 μg), TGF‐β2 (100‐35B‐10 μg) and TGF‐β3 (100‐36E‐10 μg) proteins were purchased from PeProtech Inc (Peprotech). All TGF‐β isoforms were dissolved according to the manufacturer's protocol and the cell viability after TGF‐β isoforms treatment was determined by a methyl thiazolyl tetrazolium (MTT, Amresco) assay.¹⁴ Sofosbuvir (HY‐15005), TβRI/II inhibitor LY2109761 (HY‐12075) and TβRI/ALK5 inhibitor RepSox (HY‐13012) were purchased from MedChem Express (MCE). The following primary antibodies were used: anti‐HCV core (ab2740, Abcam), anti‐NS3 (ab13830, Abcam), anti‐β‐Actin (3700S, Cell Signaling Technology), anti‐TGF‐β1 (ab179695, Abcam), anti‐TGF‐β3 (ab15537, Abcam), anti‐TβRI (ab121024, Abcam), anti‐SMAD2/3 (8685T, Cell Signaling Technology) and corresponding secondary antibodies (ZSGB‐BIO). The siRNAs for TβRI (sc‐40222) and for SMAD2/3 (sc‐37238) and the negative control siRNA‐A (sc‐37007) were purchased from Santa Cruz Biotechnology.

Zou L., Li J., Li H., Tan J., Wang M., Liu N., Gao R., Yan H., Wang X., Dong B., Li Y, & Peng Z. (2021). TGF‐β isoforms inhibit hepatitis C virus propagation in transforming growth factor beta/SMAD protein signalling pathway dependent and independent manners. Journal of Cellular and Molecular Medicine, 25(7), 3498-3510.

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Murine T Cell Polarization Assay

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Anti-mouse CD3e (100340), anti-mouse CD28 (102116), recombinant mouse IL-2 (575406), recombinant mouse IL-6 (575702), recombinant mouse TGF-β (763102), recombinant mouse IL-12 (577004), recombinant mouse IL-23 (589002), Brilliant Violet 421-conjugated anti-mouse CD4 (100443), PerCP anti-mouse CD8a (100731), PE-conjugated anti-mouse/human CD44 (103008), APC-conjugated anti-mouse CD62L (104412), PE/Cyanine7-conjugated anti-mouse IFN-γ (505826), Brilliant Violet 421-conjugated anti-mouse IL-17A (512321), PE-conjugated anti-mouse IL-17A (506904), Alexa Fluor^® 647 anti-mouse/rat/human FOXP3 (320014), FITC anti-mouse CD11c (117306), Brilliant Violet 421™ anti-mouse I-A/I-E (107620), PE/Cy7 anti-mouse CD86 (105014), and PE anti-mouse F4/80 (123110) were purchased from Biolegend (San Diego, CA, USA). Anti-phospho-PI3 Kinase p85 (Tyr458)/p55 (Tyr199) (4228S), anti-PI3 Kinase p85 (4292S), anti-phospho-AKT (Ser473) (D9E) XP^® Rabbit mAb (4060S), anti-AKT (9272S), anti-PKM2 (4053S) and anti-β-Actin (3700S) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA); anti-PGK1 (A12686), anti-ENO1 (A11448), and anti-LDHA (A1146) antibodies were got from Abclonal (Wuhan, China); anti-HK2 (22029-1-AP) and anti-Glut1 (21829-1-AP) antibodies were obtained from Proteintech (Wuhan, China); and 740 Y-P (HY-P0175) was ordered from Medchem Express (Shanghai, China).

Zou Y., Zhang J., Sun F., Xu Q., Chen L., Luo X., Wang T., Zhou Q., Zhang S., Xiong F., Kong W., Yang P., Yu Q., Liu S, & Wang C.Y. (2024). Fluvoxamine inhibits Th1 and Th17 polarization and function by repressing glycolysis to attenuate autoimmune progression in type 1 diabetes. Molecular Medicine, 30, 23.

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Western Blot Analysis of Osteoclast Proteins

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Osteoclasts from control or Lyz2^Cre/WT;Nfatc2^Δ/Δ mice were extracted in buffer containing 25 mM Tris-HCl (pH 7.5), 150 mM NaCl, 5% glycerol, 1 mM EDTA, 0.5% Triton X-100, 1 mM sodium orthovanadate, 10 mM NaF, 1 mM phenyl methyl sulfonyl fluoride and a protease inhibitor cocktail (all from Sigma Aldrich, St. Louis, MO). Quantified total cell lysates (50 μg of total protein) were separated by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) in 8% polyacrylamide gels and transferred to Immobilon-P membranes (Millipore, Billerica, MA). The blots were probed with anti-NFATc2 (4389) and anti-β-Actin (3700) antibodies, both from Cell Signaling Technology (Danvers, MA), and anti-NFATc1 (556602; BD Bioscience, San Jose, CA) antibodies. The blots were exposed to either anti-rabbit IgG or anti-mouse IgG conjugated to horseradish peroxidase (Sigma-Aldrich) and incubated with a chemiluminescence detection reagent (BioRad). The bands were detected by ChemiDoc^™ XSR+ molecular imager (BioRad) with Image Lab^™ software (version 5.2.1) [Zanotti et al., 2013 (link)].

Yu J., Zanotti S., Schilling L, & Canalis E. (2018). NUCLEAR FACTOR OF ACTIVATED T CELLS 2 IS REQUIRED FOR OSTEOCLAST DIFFERENTIATION AND FUNCTION IN VITRO BUT NOT IN VIVO. Journal of cellular biochemistry, 119(11), 9334-9345.

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Antibody-based Autophagy Regulation Study

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Antibodies used in this study are: anti-HSP60 (sc-13115), anti-HSP90 (sc-13119) and anti-RhoGDI (sc-373723) from Santa Cruz Biotechnology; anti-LC3B (3868 for immunofluorescence and 2775 for Western blot), anti-β-actin (3700) and anti-FoxO1 (2880) from Cell Signaling; anti-CD68 (MCA1957GA) from Bio-Rad (formerly AbDSerotec). Fluorescent Alexa-coupled secondary antibodies and DAPI were from Life Technologies and HRP-coupled secondary antibodies from Cell Signaling Technology. Galanin was purchased from GeneCust and was referred to as Gal throughout this study. Bafilomycin A1, a proton-ATPase inhibitor, from Sigma-Aldrich, was used to inhibit the autophagic flux. All other chemicals were from Sigma-Aldrich unless otherwise stated.

Martinelli I., Timotin A., Moreno-Corchado P., Marsal D., Kramar S., Loy H., Joffre C., Boal F., Tronchere H, & Kunduzova O. (2021). Galanin promotes autophagy and alleviates apoptosis in the hypertrophied heart through FoxO1 pathway. Redox Biology, 40, 101866.

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Quantitative Protein Expression Analysis

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Protein lysates were isolated and concentration was determined as previously described [26 (link)]. To determine protein levels, we used the Protein Simple WES capillary western blot system (San Jose, CA, USA) according to manufacturer’s instructions. Total protein concentrations of 0.2 mg/ml was used based on prior titration. Primary antibodies were used at 1:50 dilution. The primary antibodies used were: Rabbit polyclonal anti sigma-1R NBP1–82479 from NovusBio (Centennial, CO, USA), and mouse monoclonal anti β-actin #3700 from Cell Signaling Technology (Boston, MA, USA). Anti-Rabbit and anti-mouse secondary antibodies were supplied with the WES detection module kits from Protein Simple and the manufacturer’s compass software was used for data analysis. Band images based upon the densitometry data were generated using the software for figures. The raw densitometry peaks were utilized for quantitative analyses.

Motawe Z.Y., Farsaei F., Abdelmaboud S.S., Cuevas J, & Breslin J.W. (2020). Sigma-1 Receptor Activation-Induced Glycolytic ATP Production and Endothelial Barrier Enhancement. Microcirculation (New York, N.Y. : 1994), 27(6), e12620.

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Antibody Validation for Adipogenesis Study

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Anti-G9a (PP-A8620A-00; mouse monoclonal) and anti-GLP (PP-B0422-00; mouse monoclonal) antibodies were purchased from R&D Systems (Minneapolis, MN, USA). Anti-H3K9me1 (ab8896; rabbit polyclonal) and anti-STAT5A (ab32043; rabbit monoclonal) antibodies were purchased from Abcam (Cambridge, UK). Anti-Adiponectin (PA1-054; rabbit polyclonal) antibody was purchased from Thermo Scientific (Waltham, MA, USA). Anti-H3K9me2 (9753; rabbit polyclonal), anti-Histone H3 (14269; mouse monoclonal), anti-HSL (4107; rabbit polyclonal), anti-Perilipin (3470; rabbit polyclonal), and anti-β-actin (3700; mouse monoclonal) antibodies were purchased from Cell Signaling (Danvers, MA, USA).

Able A.A., Richard A.J, & Stephens J.M. (2023). TNFα Effects on Adipocytes Are Influenced by the Presence of Lysine Methyltransferases, G9a (EHMT2) and GLP (EHMT1). Biology, 12(5), 674.

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Western Blotting of Topoisomerases

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Antibodies used were as follows: anti–β-actin (3700; Cell Signaling); anti-TOP2A (ab52934; Abcam), anti-TOP2B (ab58442; Abcam), anti-TOP1 (A302-589A; Bethyl), anti-FLAG-HRP (A8592; Sigma). Gradient gels were used according to the molecular weight of the proteins to be evaluated, followed by wet transfer on polyvinylidene fluoride membranes.

Rialdi A., Campisi L., Zhao N., Lagda A.C., Pietzsch C., Ho J.S., Martinez-Gil L., Fenouil R., Chen X., Edwards M., Metreveli G., Jordan S., Peralta Z., Munoz-Fontela C., Bouvier N., Merad M., Jin J., Weirauch M., Heinz S., Benner C., van Bakel H., Basler C., García-Sastre A., Bukreyev A, & Marazzi I. (2016). Topoisomerase 1 inhibition suppresses inflammatory genes and protects from death by inflammation. Science (New York, N.Y.), 352(6289), aad7993.

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Exploring Cell Line Manipulation Techniques

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Human malignant Hep3B, HepG2, SNU-387 and Li-7 cells were provided by the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Human normal MIHA and malignant MHCC-97H, LM3, Huh7, SNU-368, and SNU-739 cells were provided by the BeNa Culture Collection (Henan, China). Cell culture was carried out with DMEM (Gibco, USA) supplied with 10% (v/v) fetal bovine serum (FBS; Gibco), 100µg/mL streptomycin (Gibco) and 100 U/mL penicillin (Gibco) in a humid incubator (5% CO 2 , 37 °C). ASRGL1 shRNA and NC shRNA lentiviral vectors containing the green fluorescent protein (GFP) expression sequence were provided by Genechem Company (China). The target sequence of ASRGL1 shRNA was 5′-CGCAGTCCAGTGTATAGCAAA-3′. The plasmids with stable overexpression of ASRGL1 (ASRGL1-OE) or CDK1 (CDK1-OE) were produced by the Shanghai GeneChem Co., Ltd (GeneChem, China). The empty vector was served as a control. Anti-ASRGL1 antibodies were provided by Santa Cruz Biotechnology (sc-130472). Anti-p53 (2527) and anti-β-actin (3700) antibodies were provided by Cell Signaling Technology. Anti-CyclinB1 (55004-1-AP), anti-CyclinE1 (11554-1-AP), and anti-CDK2 (10122-1-AP) antibodies were provided by Proteintech. Anti-Bub1 antibodies (ab195268) were provided by Abcam. Anti-CDK1 (D190678) and anti cdc20 (D225920) antibodies were from BBI Life Sciences Corporation.

Wang X., Wang Y., Yang L., Yuan J., Shen W., Zhang W., Wang J, & Tao K. (2023). ASRGL1 downregulation suppresses hepatocellular carcinoma tumorigenesis in a CDK1-dependent manner. Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver, 55(7).

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