Anti alpha tubulin

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-alpha-tubulin is a primary antibody that specifically recognizes the alpha-tubulin subunit of the microtubule cytoskeleton. It can be used in various applications, such as Western blotting, immunohistochemistry, and immunocytochemistry, to detect and visualize alpha-tubulin in biological samples.

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6 protocols using anti alpha tubulin

Apoptosis Signaling Pathway Analysis

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OAW42 cells were grown up to 70% confluency, treated with DMSO, only MRE or MRE in combination with cisplatin/paclitaxel for 24 h. Then cells were harvested. The whole cell lysate of individual treatment groups were prepared. Expression of Caspase 9, Caspase 8, Caspase 3 and PARP1 was carried out through western blot, following established protocol.¹⁸ (link) Antibodies were procured from Cell signaling Technology, Inc. and were used in the following dilutions: anti-caspase 9 (Mouse monoclonal, 1:1000 dil. Cat No: 9508), anti-caspase 8 (Mouse monoclonal, 1:1000 dil. Cat No. 9746), anti-caspase 3 (Rabbit polyclonal, 1:1000 dil. Cat No. 9662), anti-cleaved caspase 3 (Rabbit Polyclonal, 1:1000 dil. Cat No. 9661), anti-PARP1 (Rabbit polyclonal, 1:1000 dil. Cat No. 9541). Anti-alpha tubulin (mouse monoclonal, 1:1000 dil.) was procured from Santa Cruz Biotechnology Inc, Cat No: sc8035). Following secondary antibodies were used: HRP conjugated anti-rabbit IgG raised in Goat (Cat No. A0545, Sigma, 1:8000 dil.) and HRP conjugated anti-mouse IgG raised in Rabbit (Cat No. A9044, Sigma, 1:10,000 dil.).

Ghosh A., Roychowdhury T., Nandi R., Maiti R., Ghosh N.N., Molla S.A., Mukhopadhyay S., Prodhan C., Chaudhury K., Das P., Sarkar N.K., Chattopadhyay S., Bhattacharya R., Bose C.K, & Maiti D.K. (2021). Inhibitory role of a smart nano-trifattyglyceride of Moringa oleifera root in epithelial ovarian cancer, through attenuation of FSHR - c-Myc axis. Journal of Traditional and Complementary Medicine, 11(6), 481-492.

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Protein Immunoblot Analysis of EMT6 and 4T1 Cells

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EMT6 or 4T1 cells were harvested for protein lysates for immunoblotting as described in [31] (link). List of antibodies: anti-collagen I (ab34710, abcam), anti-beta-catenin (sc-7963, Santa Cruz Biotechnology), anti-alpha-tubulin (sc-8935, Santa Cruz Biotechnology), anti-phospho-LRP6 (#2568, Cell Signaling Technology), and anti-LRP6 (#3395, Cell Signaling Technology). Densitometry was determined by the ImageJ program.

Willis C.M, & Klüppel M. (2014). Chondroitin Sulfate-E Is a Negative Regulator of a Pro-Tumorigenic Wnt/Beta-Catenin-Collagen 1 Axis in Breast Cancer Cells. PLoS ONE, 9(8), e103966.

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Antibody-Based Protein Interaction Analysis

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The following antibodies were used: anti-GFP (Santa Cruz, sc-9996 for coimmunoprecipitation; Abcam, ab32146 for immunoblots), anti-FLAG horseradish peroxidase-conjugated (Sigma, A8592), anti-p-SEK1/MKK4 (Cell Signaling, CST-9151), anti-GST (Santa Cruz, sc-138), anti-alpha-tubulin (Santa Cruz, sc-53030), anti-DLK (ThermoFisher Scientific, PA5-32173 for coimmunoprecipitation and immunohistochemistry; Antibodies Incorporated, 75-355 for immunoblot), anti-GAPDH (Santa Cruz, sc-32233), anti-LC3A/B (Cell Signaling, CST-12741), anti-HA (Abcam, ab9110), anti-Xpress (ThermoFisher Scientific, R910-25), anti-SUMO2/3 (Abcam, ab3742), anti-FLAG (Cell Signaling, 14793S), anti-BRN3A Alexa Fluor 594 (Santa Cruz, sc-8429 AF594), and anti-beta III tubulin (Abcam, ab41489). We dissolved all chemicals in dimethyl sulfoxide (Sigma, D8418-250ML) except for vincristine (Sigma, V8879), which was dissolved in methanol. Controls were treated with dimethyl sulfoxide as vehicle or methanol in the case of vincristine controls. We used vincristine at 200 nM, bafilomycin (Sigma, B1793) at 100 nM, caspase inhibitor (Sigma, 400012) at 1 and 5 μM, pan-caspase inhibitor (R&D systems, FMK001) at 10 and 50 μM, and MG-132 (Sigma, M7449) at 10 μM.

Lee B., Oh Y., Cho E., DiAntonio A., Cavalli V., Shin J.E., Choi H.W, & Cho Y. (2022). FK506-binding protein-like and FK506-binding protein 8 regulate dual leucine zipper kinase degradation and neuronal responses to axon injury. The Journal of Biological Chemistry, 298(3), 101647.

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Cell Lysis and Western Blotting

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Cells were lysed at 4 °C using ice cold lysis buffer containing 30 mM Tris/HCl pH 6.7, 5% glycerol, 2.5% β-mercaptoethanol, and 1% SDS. Protein extracts were analyzed by SDS-PAGE and western blotting. Enhanced chemiluminescence (ECL) signals were detected as described before [38 (link)]. The following antibodies were used for western blot: anti-AXL C89E7 (Cell Signaling Technology, Boston, MA, USA), anti-CD68 (Cell Signaling Technology, Boston, MA, USA), anti-DICER1 clone D38E7 (Cell Signaling Technology, Boston, MA, USA), anti-SPRY4 (GeneTex, Irvine, CA, USA), pan-AGO clone 2A8 (Millipore, Overijse, Belgium), anti-vinculin E1E9V XP (Cell Signalling Technology, Boston, MA, USA), anti-alpha-tubulin (Santa Cruz, Heidelberg, Germany). HRP-labelled secondary antibodies were purchased from Cell Signaling Technology (Boston, MA, USA).

Kozar I., Philippidou D., Margue C., Gay L.A., Renne R, & Kreis S. (2021). Cross-Linking Ligation and Sequencing of Hybrids (qCLASH) Reveals an Unpredicted miRNA Targetome in Melanoma Cells. Cancers, 13(5), 1096.

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Murine Asthma Model Protocol

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Luteolin, picrotoxin, ovalbumin and the PAS staining kit and other chemical reagents were purchased from Sigma (Sigma-Aldrich, St. Louis, MO, USA). Anti-Muc5ac and anti-alpha-tubulin antibodies were purchased from Santa Cruz Biotechnology (sc-21701, sc-12462-R, Dallas, TX, USA). Horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from Pierce Biotechnology (Thermo Fisher Scientific, Rockford, IL, USA). IL-4, IL-5 and IgE ELISA kits were purchased from BD Biosciences (San Diego, CA, USA). The IL-13 kit was purchased from R&D Systems (Minneapolis, MN, USA). OVA for intraperitoneal (i.p.) injection was adsorbed to aluminum hydroxide adjuvant (Santa Cruz, Dallas, TX, USA) at a ratio of 50 μg to 2 mg in 200 μl PBS. OVA for intratracheal (i.t.) injection was dissolved in saline at a final concentration of 2.5 mg/ml. Luteolin (Sigma-Aldrich, St. Louis, MO, USA) was prepared in DMSO and diluted with saline. picrotoxin (Sigma-Aldrich, St. Louis, MO, USA) was prepared in DMSO and diluted with saline. Saline with 0.5% DMSO was used as a vehicle in control groups. The final concentration of DMSO in all reaction mixtures was less than 0.5%.

Shen M.L., Wang C.H., Lin C.H., Zhou N., Kao S.T, & Wu D.C. (2016). Luteolin Attenuates Airway Mucus Overproduction via Inhibition of the GABAergic System. Scientific Reports, 6, 32756.

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Antibodies for Mitochondrial Protein Analysis

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The following primary antibodies were used for Western blots: anti-Fis1 (LuBioScience GmbH, 10956-1-AP, diluted 1:2000), anti-Mff (Life Technologies, PA5-52765, diluted 1:500-1:1000), anti-Inf2
(Sigma-Aldrich HPA000724, diluted 1:2000), anti-alpha-Tubulin (Santa Cruz sc-5286, diluted 1:2000).

Kleele T., Rey T., Winter J., Zaganelli S., Mahečić D., Lambert H.P., Ruberto F.P., Nemir M., Wai T., Pedrazzini T., & Manley S. (2020). Distinct molecular signatures of fission predict mitochondrial degradation or proliferation.

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