β tubulin antibody

Manufactured by Merck Group

Sourced in United States

The β-tubulin antibody is a laboratory research tool used to detect and quantify the β-tubulin protein in various cell and tissue samples. β-tubulin is a key structural component of microtubules, which are essential for various cellular processes such as cell division, intracellular transport, and cell motility. The antibody can be used in techniques like Western blotting, immunocytochemistry, and immunohistochemistry to analyze the expression and distribution of β-tubulin in different experimental systems.

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Anti-β-tubulin monoclonal antibody Anti-β-tubulin antibody Anti-β-tubulin antibody (T8328, Mouse monoclonal anti-β-tubulin antibody (T4026 Anti-β-tubulin III antibody β-Tubulin I antibody Anti‐β‐Tubulin antibody (T5293; Mouse anti-β-tubulin III monoclonal antibody (clone SDL.3D10, Mouse monoclonal anti-β-III-tubulin antibody Rabbit anti-β-tubulin antibody

31 protocols using β tubulin antibody

Quantifying TREM2 and Microglial Activation

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For western-blots we used rabbit polyclonal antibodies against the synthetic peptide within 29–59 N-terminal aa’ of human TREM2 (1:200, Abcam, ab175262, Cambridge, MA) (Ma et al., 2016 (link); Raha et al., 2016 ) and against to the C-terminus of Iba1 (1:500, Wako, Osaka, Japan). TREM2 antibody recognized non-glycosylated/immature (~ 25 kDa) and glycosylated/mature (~ 50 kDa) form (Fig. S1). Loading control was a β-tubulin antibody (1:1000, Millipore, Billerica, MA) (Mufson et al., 2012 (link); Perez et al., 2015 (link)). Aβ plaques and microglia quantitation employed a mouse monoclonal IgG antibody against human APP/Aβ (6E10) (1:2000, BioLegend, San Diego, CA) and a rabbit polycloncal antibody against Iba1 (1:1000, Wako), respectively.

Perez S.E., Nadeem M., He B., Miguel J.C., Malek-Ahmadi M.H., Chen K, & Mufson E.J. (2017). Neocortical and hippocampal TREM2 protein levels during the progression of Alzheimer’s disease. Neurobiology of aging, 54, 133-143.

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Bryostatin-1 and Picolog Analog Synthesis

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Bryostatin-1 (Cat# 2383) was purchased from Tocris Bioscience (Minneapolis, MN). The Erk inhibitor PD98059 (Cat# P215) was purchased from Sigma-Aldrich (St. Louis, MO). P44/42 (Erk1/2) rabbit monoclonal antibody (Cat# 4695) and phosphoP44/42 MAPK (Erk1/2) (Thr202/Tyr204) rabbit polyclonal antibody (Cat# 9101) were purchased from Cell Signaling Technology (Danvers, MA). Human/mouse caspase-8 antibody (Cat# AF1650) was purchased from R&D Systems (Minneapolis, MN). β-actin (AC-15) antibody (Cat#sc-69879) was purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). β-tubulin antibody (Cat# 04–1049) was purchased from Millipore (Temecula, CA). The design and synthesis of the synthetic analog of Bryostatin-1, picolog, were conducted according to published methods (Wender et al., 2014 ). The chemical formula of Bryostatin-1 and picolog are presented in Fig.1. Picolog was synthesized by function-oriented synthesis using Bryostatin-1as therapeutic lead in a step-economical fashion. Picolog was found to be the most potent new class of ‘B-Ring’ analog of Bryostatin-1 in terms of PKC activity (Wender et al., 2015 (link)). Bryostatin-1 and picolog were dissolved in DMSO (50µM) and aliquots were stored at −20 °C. Stock solutions were dissolved in culture medium at different concentrations just before each experiment.

Khan T.K., Wender P.A, & Alkon D.L. (2017). Bryostatin and its synthetic analog, picolog rescue dermal fibroblasts from prolonged stress and contribute to survival and rejuvenation of human skin equivalents. Journal of cellular physiology, 233(2), 1523-1534.

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Western Blot Analysis of ER Stress Proteins

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Thirty micrograms of different protein fractions from rat tissues were subjected to 10% SDS-polyacrylamide gel electrophoresis. Proteins were transferred to Immobilon polyvinylidene difluoride transfer membranes (Millipore, Burlington, MA, USA) and blocked for 1 h at room temperature with 5% non-fat milk solution in 0.1% Tween-20-Tris-buffered saline (TBS). Membranes were then incubated overnight with the primary antibody in 0.1% Tween-20-TBS with 5% bovine serum albumin (BSA) at 4 °C. Detection was achieved using the enhanced chemiluminescence (ECL) kit for horseradish peroxidase (HRP) (Amersham GE Healthcare Europe GmbH, Barcelona, Spain). To confirm the uniformity of protein loading, the blots were incubated with β-actin or β-tubulin antibody (Sigma-Aldrich, Saint Louis, MO, USA) as a control. The size of the detected proteins was estimated using protein molecular-mass standards (Invitrogen, Life Technologies). Primary antibodies for phosphor- and total-IRE1, ATF6α, phosphor- and total-PERK, and XBP1s proteins were obtained from Santa Cruz Biotechnologies (Dallas, TX, USA) and AbCam (Cambridge, UK) as described previously [32 (link)].

Rodrigo S., Panadero M.I., Fauste E., Rodríguez L., Roglans N., Álvarez-Millán J.J., Otero P., Laguna J.C, & Bocos C. (2019). Effects of Maternal Fructose Intake on Perinatal ER-Stress: A Defective XBP1s Nuclear Translocation Affects the ER-stress Resolution. Nutrients, 11(8), 1935.

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Curcumin Modulates Steroidogenesis In Vitro

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Curcumin was purchased from Cayman Chemical (item No. 81025). Cell culture supplies were obtained from Gibco (Thermo Fisher Scientific) and plasticware from BD Biosciences. [³H]-thymidine (20 Ci/mmol) and [^1,2,6,7-3H]-progesterone were purchased from New England Nuclear Corporation (North Billerica, MA, USA). StAR antibody (item No. FL-285) was from Santa Cruz Biotechnology Inc., β-tubulin antibody (item No. 05-661-I) was from Sigma-Aldrich, and peroxidase-conjugated anti-mouse (item No. PI-2000-1) and anti-rabbit (item No. PI-1000-1) IgG antibodies from Vector Labs (Burlingame, CA, USA). WST-1, Trypan blue, propidium iodide, sodium citrate, RNase A, activated charcoal, dextran, and progesterone antibody were purchased from Sigma-Aldrich.

Raices T., Luisa Varela M., Abiuso A.M., Pereyra E.N., Mondillo C., Pignataro O.P, & Riera M.F. (2023). Curcumin effects on Leydig cell functions and potential therapeutic uses. Endocrine Oncology, 3(1), e220075.

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Investigating ER Stress Signaling Pathways

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Antibodies used in this study include the following: GRP78 antibody (N-20 and C-20, Santa Cruz Biotechnology, Santa Cruz, CA, USA), Phosphos-IRE1α (Ser724) antibody (Abcam, Cambridge, UK); E-cadherin (Stressgen, Victoria, Canada); STAT3, Phospho-STAT3 (Tyr705), JAK2, Phospho-JAK2 (Tyr1007/1008), CHOP, Caspase-3, IRE1α, and PARP antibody (Cell Signaling Technology, Beverley, MD, USA); and β-tubulin antibody (Sigma-Aldrich, Steinheim, Germany). Tunicamycin was from Sigma-Aldrich. Dulbecco’s Modified Eagles Medium (DMEM), fetal bovine serum (FBS), and Geneticin (G418) were purchased from HyClone (Logan, UT, USA). STAT3 specific inhibitor benzoic acid (2-Hydroxy-4-(((4-methylphenyl)sulfonyloxy)acetyl)amino)-benzoic acid, NSC74859), human STAT3/shRNA, and control shRNA lentiviral particles were obtained from Santa Cruz Biotechnology.

Yao X., Liu H., Zhang X., Zhang L., Li X., Wang C, & Sun S. (2015). Cell Surface GRP78 Accelerated Breast Cancer Cell Proliferation and Migration by Activating STAT3. PLoS ONE, 10(5), e0125634.

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Investigating MEK/ERK Signaling Pathway

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Plasmids pGL3-basic and pRL-TK, as well as the MEK1/2-specific inhibitor U0126, were purchased from Promega. Antibodies against Sp1 (rabbit monoclonal antibody, 1:1000 dilutions), c-Jun (rabbit polyclonal antibody, 1:500 dilutions), c-Fos (rabbit polyclonal antibody, 1:500 dilutions), ERK1/2 (rabbit polyclonal antibody, 1:1000 dilutions), and β-actin (mouse monoclonal antibody, 1:1000 dilutions) were purchased from Santa Cruz Biotechnology. Antibodies against p-ERK1/2 (Thr202/Tyr204) (rabbit monoclonal antibody, 1:1000 dilutions), T567 ezrin (rabbit monoclonal antibody, 1:1000 dilutions) were purchased from Cell Signaling (Beverly, MA) and the ezrin antibody was purchased from Neomarker (mouse monoclonal antibody, 1:500 dilutions). TPA, dimethyl sulfoxide (DMSO), and β-tubulin antibody (mouse monoclonal antibody, 1:1000 dilutions) were purchased from Sigma. All other reagents were of analytical reagent grade.

Zhang X.D., Xie J.J., Liao L.D., Long L., Xie Y.M., Li E.M, & Xu L.Y. (2015). 12-O-Tetradecanoylphorbol-13-Acetate Induces Up-Regulated Transcription of Variant 1 but Not Variant 2 of VIL2 in Esophageal Squamous Cell Carcinoma Cells via ERK1/2/AP-1/Sp1 Signaling. PLoS ONE, 10(4), e0124680.

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Western Blot Analysis of Tight Junction Proteins

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Protein concentration from homogenized microvessels was measured using BCA^™ Protein Assay Kit (Thermo Scientific, Rockford, IL). Equal amounts of protein lysates (20 μg) were loaded and separated on a 4–20% sodium dodecyl sulphate-polyacrylamide gels (SDS-PAGE), transferred using nitrocellulose membranes (Bio-Rad Laboratories, Hercules, CA) and analyzed by Western Blotting. nitrocellulose membranes were blocked with 3% bovine serum albumin (BSA) dissolved with Tris-buffered saline containing 0.1% Tween-20 (TBS-T), and incubated with the respective antibodies overnight at 4 °C. Occludin and ZO-1 antibodies were purchased from Invitrogen (Camarillo, CA). Claudin-5 antibody was purchased from Millipore (Temecula, CA). β-tubulin antibody was obtained from Sigma-Aldrich (St. Louis, MO). All antibodies were diluted with 3% BSA in TBS-T buffer at 1:1000 except for β-tubulin which was diluted 1:20,000. Secondary antibodies were purchased from LI-COR biosciences (Lincoln, NE) and incubated at room temperature for 1 hour at 1:20,000 concentrations. Following washing with TBS-T blots were imaged on the LI-COR Odyssey® CLx scanner and analyzed using Image Studio software provided with the instrument.

Wolff G., Davidson S.J., Wrobel J.K, & Toborek M. (2015). Exercise maintains blood-brain barrier integrity during early stages of brain metastasis formation. Biochemical and biophysical research communications, 463(4), 811-817.

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Metformin effects on lipid metabolism

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Metformin (1,1-dimethylbiguanide hydrochloride) was purchased from Sigma-Aldrich (St. Louis, MO, USA). The [9,10(n)-³H] palmitic acid was purchased from NEN (Boston, MA, USA). Phosphorylated ACC (Ser79), phosphorylated p70S6K (Thr389), total ACC, and total p70S6K antibodies were acquired from Cell Signaling Technology (Danvers, MA, USA). The androgen receptor (AR) antibody was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA), claudin 11 antibody was obtained from Invitrogen (Thermo Fisher Scientific, Waltham, MA, USA), ZO-1 antibody was acquired from Zymed Laboratories (Thermo Fisher Scientific), and β-tubulin antibody was purchased from Sigma-Aldrich. Tissue culture media and all other drugs were acquired from Sigma-Aldrich. Sprague–Dawley rats (Rattus norvegicus) were obtained from the Central Animal Facility of the Department of Veterinary Sciences (Universidad de Buenos Aires, Argentina).

Rindone G.M., Dasso M.E., Centola C.L., Sobarzo C.M., Galardo M.N., Meroni S.B, & Riera M.F. (2024). Effect of Metformin on Sertoli Cell Fatty Acid Metabolism and Blood–Testis Barrier Formation. Biology, 13(5), 330.

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Ethanol Self-Administration Protocol

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Ethanol solutions (v/v) obtained by dilution of ethanol (95%; U.S. Pharmacopeia National Formulary, 1995 ) with tap water were freshly prepared before every session of self-administration. xCT polyclonal antibody was from Abnova (Taipei, Taiwan). β-tubulin antibody was purchased from Sigma-Aldrich (St. Louis, MO, USA). Both anti-mouse and anti-rabbit IgG-coupled horseradish peroxidase antibodies as well as enhanced chemiluminescence (ECL) reagents were from Cell Signaling Technology (USA). All other reagents were of the highest purity grade commercially available.

Peana A.T., Muggironi G, & Bennardini F. (2014). Change of cystine/glutamate antiporter expression in ethanol-dependent rats. Frontiers in Neuroscience, 8, 311.

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Targeted Cancer Therapies Evaluation Protocol

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AZD9291, gefitinib, afatinib, and crizotinib were purchased from Selleckchem. SHR‐A1403, the naked anti‐c‐Met monoclonal antibody c‐Met mAb and free toxin SHR152852, were provided by Jiangsu Hengrui Medicine Co. Ltd.31 DyLight 488 N‐hydroxysuccinimide (NHS) ester was purchased from Thermo‐Fisher Scientific. Sulforhodamine B was purchased from Sigma‐Aldrich.
Antibodies against EGFR, phospho‐EGFR (Tyr1173), c‐Met, phospho‐c‐Met (Tyr1234/1235), STAT3, phospho‐STAT3 (Tyr705), AKT, phospho‐AKT (Ser473), ERK1/2, phospho‐ERK1/2 (Thr202/Tyr204), and GAPDH were purchased from Cell Signaling. β‐Tubulin antibody was purchased from Sigma‐Aldrich.

Tong M., Gao M., Xu Y., Fu L., Li Y., Bao X., Fu H., Quan H, & Lou L. (2019). SHR‐A1403, a novel c‐mesenchymal‐epithelial transition factor (c‐Met) antibody‐drug conjugate, overcomes AZD9291 resistance in non‐small cell lung cancer cells overexpressing c‐Met. Cancer Science, 110(11), 3584-3594.

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