Tubulin b512

Manufactured by Merck Group

Sourced in United States

Tubulin #B512 is a laboratory product manufactured by Merck Group. It is a purified protein that plays a critical role in the structure and function of microtubules, which are essential components of the cytoskeleton in eukaryotic cells.

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Anti-α-tubulin clone B512 (23 citations) Anti-tubulin (B512, (8 citations) Mouse anti-α–tubulin B512 (7 citations) Mouse monoclonal anti-α-tubulin B512 (4 citations) Mouse monoclonal anti-Tubulin B512 (2 citations) Anti-Tubulin B512 mouse monoclonal antibody (2 citations)

6 protocols using tubulin b512

Epithelial-Mesenchymal Transition Markers

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E-cadherin #3195, Vimentin #5741, ZEB1 #3396, Claudin1 #4933, Snail #3879, β-catenin #9562, Bip #3183, CHOP #2895, eIF2α #9722 and p-eIF2α #9721 (Cell Signaling Technologies Inc. Danvers, MA 01923); Tubulin #B512 (Sigma); GAPDH #FL335 (Santa Cruz); Ubqln polyclonal was made by inoculating rabbits with a peptide specific to Ubqln1 (Yenzym Antibodies LLC); Alexa Fluor 488 goat anti-rabbit IgG #A11034 (Molecular Probes, Invitrogen detection technologies, Eugene, OR. USA); Alexa Fluor 568 Phalloidin #A12380 (Life technologies Eugene, OR. USA).

Shah P.P., Lockwood W.W., Saurabh K., Kurlawala Z., Shannon S.P., Waigel S., Zacharias W, & Beverly L.J. (2014). Ubiquilin1 Represses Migration and Epithelial to Mesenchymal Transition of Human Non-small Cell Lung Cancer Cells. Oncogene, 34(13), 1709-1717.

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Antibody Characterization for Research

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The following rabbit antibodies were used: Flag (Sigma-Aldrich, St Louis, MO, United States), c-Myc-HRP (Miltenyi Biotec, North Rhine-Westphalia, Germany); RPL11 (D1P5N), Phospho-4EBP1 (Thr70), 4EBP1, S6 ribosomal protein (5G10), Phospho-S6 ribosomal protein (Ser235/236) and TSC2 (Cell Signaling, Danvers, MA, United States) and HA-HRP (Thermo Fisher Scientific, Waltham, Massachusetts, United States). Mouse monoclonal antibodies used were: HA, c-Myc, RPS23 (1E3) and Tubulin (B512) (Sigma-Aldrich, St Louis, MO, United States); eIF4E (BD Biosciences, Franklin Lakes, New Jersey, United States); p72 (1BC11), p72 (18BG3), and p150 (17AH2) (Ingenasa, Madrid, Spain); Cullin4B and RPS23 (Abcam, Cambridge, United Kingdom) and p30 (a gift from J.M. Escribano, Algenex, Madrid, Spain). Horseradish peroxidase-conjugated anti-rabbit and anti-mouse secondary antibodies were from Sigma-Aldrich (St Louis, MO, United States), and Alexa-Fluor−488, −594, and −647 conjugated anti-rabbit and anti-mouse antibodies were from Thermo Fisher Scientific (Waltham, Massachusetts, United States).

Barrado-Gil L., Del Puerto A., Muñoz-Moreno R., Galindo I., Cuesta-Geijo M.Á., Urquiza J., Nistal-Villán E., Maluquer de Motes C, & Alonso C. (2020). African Swine Fever Virus Ubiquitin-Conjugating Enzyme Interacts With Host Translation Machinery to Regulate the Host Protein Synthesis. Frontiers in Microbiology, 11, 622907.

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Immunoblotting Analysis of IGF/Insulin Signaling

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IGF1R-beta (CST#3027), IGF1R Beta XP (CST#9750), p-IGF1R beta (CST#3918), INSR (CST#3025), IGF2R (CST#14364), AKT (CST#9272), p-AKT (CST#9271); Tubulin #B512 (Sigma); GAPDH (SantaCruz#FL335); Actin (Sigma#A5316), Ubiquilin1 (CST#14526); Anti-FLAG Affinity Gel (Sigma#A2220), INSR (CST#3025), FLAG polyclonal (Sigma# F7425).

Kurlawala Z., Dunaway R., Shah P.P., Gosney J.A., Siskind L.J., Ceresa B.P, & Beverly L.J. (2017). Regulation of Insulin-like Growth Factor Receptors by Ubiquilin1. The Biochemical journal, 474(24), 4105-4118.

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Plasmid Expression and Antibody Detection

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Plasmids expressing ^FTALLC, ^FTJTO, or untagged ALLC in the pCMV1 or pDEST40 vector were described previously (Cooley et al., 2014 (link)). ^FTBiP and ERdj3 overexpression plasmids were used as described previously (Genereux et al., 2015 (link)). The GRP94 overexpression plasmid was prepared using GRP94.pDONR223 (Addgene; Cat #82130), which was recombined into pDEST40 using Gateway cloning according to the manufacturers protocol. Primary antibodies were acquired from commercial sources and used at the indicated dilutions in Antibody Buffer (50 mM Tris [pH 7.5], 150 mM NaCl supplemented with 5% BSA and 0.1% NaN₃). Mouse monoclonal antibodies were used to detect KDEL (1:1000, Enzo Life Sciences), M2 anti-FLAG (1:500, Sigma Aldrich), Tubulin [B-5-1-2] (1:4000,Sigma), BiP/GRP-28 (1:500, Santa Cruz Biotechnology), β-actin (1:10000, Sigma Aldrich). Polyclonal rabbit antibodies were used to detect GRP94 (1:1000, GeneTex), HYOU1 (1:1000, GeneTex), ERdj3 (DNAJB11) (1:1000, ProteinTech), PDIA4 (1:1000, ProteinTech), PDIA6 (1:1000, GenTex), PDIA3 (ERp57) (1:1000, CellSignaling).

Plate L., Rius B., Nguyen B., Genereux J.C., Kelly J.W, & Wiseman R.L. (2019). Quantitative Interactome Proteomics Reveals a Molecular Basis for ATF6-Dependent Regulation of a Destabilized Amyloidogenic Protein. Cell chemical biology, 26(7), 913-925.e4.

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Epithelial-Mesenchymal Transition Markers

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Isolation and Analysis of Cellular Proteins

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For the isolation of cellular proteins cells were lysed in RIPA lysis buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.5 % NP-40, 15 % Glycerol, supplemented with 10 µg/ml of each aprotinin, leupeptin and pepstatin as well as 0.8 µM Pefabloc (Roche, Mannheim, Germany), 1 mM NaF and 1 mM Na 3 VO 4 ). The protein concentration of the lysates was determined using Biorad Protein Assay (Biorad, Munich, Germany). Proteins were separated by SDS-PAGE and transferred to a polyvinylidenedifluorid membrane. Antigens were detected by incubation with specific primary antibodies (1:1000) followed by incubation with horseradish-peroxidase-coupled secondary antibodies (1:7500) (DAKO, Hamburg, Germany). Detection occurred via an enhanced chemoluminescence kit (Millipore Corporation, Darmstadt, Germany). List of primary antibodies: Dicer (#3363), phosphorylated ERK1/2 (#4370), ERK1/2 (#4695), phosphorylated STAT3 (#9145), phosphorylated S6RBP (#2211), S6RBP (#2217) (Cell Signalling Technology, Frankfurt/Main, Germany); STAT3 (#482) (Santa Cruz Biotechnology, Santa Cruz, CA, USA); HSC70 (1B5) (Farmingdale, NY, USA); Tubulin (B-5-1-2) (Sigma Aldrich) and REDD1 (#10638) (Proteintech, Chicago, Il, USA). Western blots were quantified using Multi Gauge software (Fujifilm Life Sciences, Düsseldorf, Germany).

Pinno J., Bongartz H., Klepsch O., Wundrack N., Poli V., Schaper F, & Dittrich A. (2016). Interleukin-6 influences stress-signalling by reducing the expression of the mTOR-inhibitor REDD1 in a STAT3-dependent manner. Cellular signalling, 28(8).

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