Mouse anti βiii tubulin

Manufactured by Merck Group

Sourced in Germany, Portugal

Mouse anti-βIII-tubulin is a laboratory reagent used in immunocytochemistry and Western blot analysis. It is a monoclonal antibody that specifically binds to the βIII-tubulin isoform, which is expressed in neuronal cells. This product can be used to detect and visualize βIII-tubulin in various biological samples.

Automatically generated - may contain errors

Lab products found in correlation

Rabbit anti tom20, by Santa Cruz Biotechnology (3 mentions) Fluoro gel mounting medium, by Electron Microscopy Sciences (3 mentions) Lsm 710 confocal microscope, by Zeiss (3 mentions) Rabbit anti rpl7a, by Abcam (2 mentions) Rabbit anti gfap, by Merck Group (2 mentions) Rabbit anti βiii tubulin, by Merck Group (2 mentions) Triton x 100, by Merck Group (2 mentions) Mouse anti actin, by Abcam (2 mentions) Mouse anti gap 43, by Thermo Fisher Scientific (2 mentions) Mouse anti tau, by Merck Group (2 mentions) Anti synaptophysin, by Santa Cruz Biotechnology (2 mentions) Anti myo6, by Merck Group (2 mentions) Anti map2, by BD (2 mentions) Anti myo6, by Santa Cruz Biotechnology (2 mentions) Anti cyto c, by BD (2 mentions) Mouse anti map2ab, by Merck Group (2 mentions) Alexa fluor 405 goat anti rabbit, by Thermo Fisher Scientific (2 mentions) Rabbit anti hb egf, by Cell Signaling Technology (2 mentions) Rabbit anti egf, by Novus Biologicals (2 mentions) Rabbit antipro egf against aa 700 800, by Novus Biologicals (2 mentions)

27 protocols using mouse anti βiii tubulin

Western Blot Analysis of Regulatory Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lysates were separated by SDS-PAGE. Proteins were transferred to nitrocellulose (pore size 0.2 µm). Membranes were blocked in blocking solution (2% (w/v) BSA, 0.1 vol% Tween 20, 0.1% (w/v) sodium azide in 1× TBS pH 7.5) for at least 1 h. Primary antibodies were diluted in blocking buffer and incubated overnight at 4 °C. The following antibodies were used in this study: polyclonal antibodies: rabbit anti-Pum2 (Abcam) 1:10,000, rabbit anti-Btz (self-made, [52 (link)]) 1:500, rabbit anti-Rpl7a (Abcam) 1:1000, rabbit anti-Rps6 1:1000, rabbit anti-phospho-Rps6 1:1000, rabbit anti-PABP1 (all Cell Signaling) 1:1000, goat anti-Vinculin (Santa Cruz) 1:200; monoclonal antibodies: mouse anti-eIF4E (BD) 1:1000, mouse anti-eIF2s1 (Cell Signaling) 1:1000, mouse anti-FMRP (gift from Utz Fischer, Würzburg) 1:1000, mouse anti-β-III-Tubulin (Sigma Aldrich) 1:10,000, and rabbit anti-phospho-mTOR (Cell Signaling) 1:1000).
Membranes were washed in PBS supplemented with 0.2 vol% Tween 20. Primary antibodies were detected using infrared dye labeled secondary anti-rabbit, anti-goat, or anti-mouse antibodies (all 1:10,000, Li-COR Biosciences). Membranes were scanned using the Li-Cor Odysey IR scanner.

Schieweck R., Schöneweiss E.C., Harner M., Rieger D., Illig C., Saccà B., Popper B, & Kiebler M.A. (2021). Pumilio2 Promotes Growth of Mature Neurons. International Journal of Molecular Sciences, 22(16), 8998.

+ Open protocol

+ Expand

Immunocytochemistry of DRG Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

DRG neurons were fixed in 4% paraformaldehyde in PBS for 15 min, permeabilized with 0.1% triton-X in PBS for 10 min, and blocked in 15% goat serum/1% BSA/0.1% triton-X in PBS. Primary and secondary antibodies were diluted in block and incubated for 1 hr each at room temperature. The following primary antibodies were used: mouse anti-βIII tubulin (1:800, Sigma), rabbit anti-TUJ1 (1:1000; Covance), chicken anti-GFP (1:500; Aves Labs), mouse anti-glutamylated tubulin GT335 (1:3000; AdipoGen), mouse anti-acetylated tubulin 6-11B-1 (1:1000; Sigma), rat anti-tyrosinated tubulin YL1/2 (1:1000; Abcam), rabbit-anti Tom20 (1:500; Santa Cruz Biotechnology). The following secondary antibodies were used at 1:500: AlexaFluor-488 anti-mouse, anti-rabbit, or anti-chicken, AlexaFluor-568 anti-rabbit, AlexaFluor-647 anti-mouse (Invitrogen), Cy3-conjugated anti-mouse, Cy5-conjugated anti-rat (Jackson ImmunoResearch). Images of fixed cells were acquired on a Nikon Eclipse E800, or when fluorescence was to be quantified, a Zeiss LSM 710 confocal microscope.

Gornstein E.L, & Schwarz T.L. (2016). Neurotoxic mechanisms of paclitaxel are local to the distal axon and independent of transport defects. Experimental neurology, 288, 153-166.

+ Open protocol

+ Expand

Western Blot Analysis of Cerebral Cortex

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fresh frozen cerebral cortex was homogenized in RIPA buffer (50 mM Tris-Cl, pH 7.5, 150 mM NaCl, 10 mM EDTA, 2 mM EGTA, 50 mM NaF, 0.5% SDS, 1% NP-40) containing a protease inhibitor cocktail (Roche Applied Science). Protein concentrations were determined by BCA (Thermo Scientific). Proteins were separated on an SDS-PAGE gel, and transferred to a nitrocellulose membrane (Bio-Rad). Membranes were blocked in TBS (100 mM Tris-Cl, pH 7.5, 150 mM NaCl) containing 0.1% Tween-20, and 5% nonfat milk solution for 1 h, then incubated with one of the following primary antibodies: mouse-anti-PSD95 (1:1000; EMD Biosciences), mouse-anti-synaptophysin (1:500; Sigma), or mouse anti-βIII-Tubulin (1:2000; Sigma) overnight at 4 °C on a rotary shaker. Membranes were washed three times with 0.1% TBS prior to addition of an anti-mouse IgG horse radish peroxidase secondary antibody (1:2000; Cell Signaling Technology). Proteins were visualized by chemiluminescence (Millipore) on a QBOX imaging system (Syngene). Densitometry analysis was performed in ImageJ (NIH) using the gel analyzer plug-in.

Dickens A.M., Yoo S.W., Chin A.C., Xu J., Johnson T.P., Trout A.L., Hauser K.F, & Haughey N.J. (2017). Chronic low-level expression of HIV-1 Tat promotes a neurodegenerative phenotype with aging. Scientific Reports, 7, 7748.

+ Open protocol

+ Expand

Immunofluorescence Staining of Cultured Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cultured cells were fixed with 4% PFA for 15 minutes at room temperature (RT) and block-permeablised using PBST-10% Normal Donkey Serum (NDS) for 1 hour at RT. Primary and secondary antibodies were incubated in 3%NDS overnight (O/N) at 4 °C and 1 hour at RT respectively at the following dilutions; mouse anti-CNPAse, (1:2000; Chemicon, Germany), rabbit anti-GFAP, goat anti-GFAP, mouse anti-βIII-tubulin, rabbit anti-βIII-tubulin (all at 1:300; Sigma-Aldrich), rabbit anti-Pax6 (1:200; Chemicon, Germany), donkey anti-sheep Alexafluor555, donkey anti-rabbit Alexafluor488/555/647 and donkey anti-mouse Alexafluor488/555/647 (all 1:1000; Invitrogen), donkey anti-chickenCy3 (Jackson Laboratories, Bar Harbour, ME). Cells were counterstained with DAPI and mounted with Slow-fade mounting media (Invitrogen). Non-specific staining was controlled by using secondary-only controls. Fluorescence was viewed using the Axioplan2 microscope (Carl Zeiss, Jena, Germany) fitted with an HBO 100 lamp (Carl Zeiss, Jena, Germany). Images were captured using an Axiocam Mrm camera and Axio Vs40 v4.5.0.0 software (Axiovision, Carl Zeiss, Jena, Germany).

Bridges C.R., Tan M.C., Premarathne S., Nanayakkara D., Bellette B., Zencak D., Domingo D., Gecz J., Murtaza M., Jolly L.A, & Wood S.A. (2017). USP9X deubiquitylating enzyme maintains RAPTOR protein levels, mTORC1 signalling and proliferation in neural progenitors. Scientific Reports, 7, 391.

+ Open protocol

+ Expand

Immunostaining Protocol for Cell Cultures

Check if the same lab product or an alternative is used in the 5 most similar protocols

Following fixation, cell permeabilisation was performed using 0.5% TritonX-100 (Sigma) for 30 min. Following 3 × 5 min washes, non-specific binding was blocked with 5% normal goat serum (Dako) in PBS for 30 min. After another wash step, primary antibodies were diluted 1:400 in PBS (mouse anti-β III-tubulin; Sigma) and incubated overnight at 4 °C. Following 3 × 10 min washes, secondary antibody, anti-mouse IgG DyLight 488 (Vector Laboratories) was diluted 1:300 in PBS and added for 90 min. Hoechst 33258 (1 μg/ml) was also added into the secondary antibody incubation to stain nuclei. Omission of a primary or secondary antibody was routinely used as a control. Incubation times for coverslips were half that for gels except for an overnight incubation in primary antibodies. Gels and coverslips were stored in PBS at 4 °C.

O’Rourke C., Hopper C., MacRobert A.J., Phillips J.B, & Woodhams J.H. (2017). Could clinical photochemical internalisation be optimised to avoid neuronal toxicity?. International Journal of Pharmaceutics, 528(1-2), 133-143.

+ Open protocol

+ Expand

Proximity Ligation Assay for Neuron Interactions

Check if the same lab product or an alternative is used in the 5 most similar protocols

The proximity ligation assay was performed according to the manufacturer’s instructions (Sigma-Aldrich). Briefly, fixed neurons were incubated with primary antibodies (SNPH-actin interaction: rabbit anti-SNPH, 1:250 and mouse anti-actin, 1:100, Abcam; SNPH-microtubules interaction: rabbit anti-SNPH, 1:100, Abcam and mouse anti-βIII-tubulin, 1:2000, Sigma-Aldrich) diluted in Duolink antibody diluent at 4°C overnight, then washed three times with Buffer A (0.01 M Tris, 0.15 M NaCl, 0.05% Tween 20) at RT for 5 min each, incubated with Duolink PLA Probe at 37°C for 1 hr, rewashed with PBS three times at RT for 5 min each, then incubated in Duolink PLA ligation solution at 37°C for 30 min, rewashed with PBS three times at RT for 5 min each, then incubated with Duolink PLA amplification solution at 37°C for 100 min, washed with 1x Buffer B (0.2 M Tris, 0.1 M NaCl pH 7.5) two times at RT for 10 min each and PBS at RT for 5 min. To counterstain for a neuronal marker, neurons were post-fixed with 4% paraformaldehyde and 4% sucrose for 10 min at RT and processed for immunocytochemistry.

Li S., Xiong G.J., Huang N, & Sheng Z.H. (2020). The crosstalk of energy sensing and mitochondrial anchoring sustains synaptic efficacy by maintaining presynaptic metabolism. Nature metabolism, 2(10), 1077-1095.

+ Open protocol

+ Expand

Immunofluorescence Staining of Neuronal Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following antibodies and dilutions were used: mouse-anti-βIII-tubulin (Sigma, #T8660, 1:400), mouse-anti-Gap-43 (Invitrogen, #33-5000, 1:50), mouse-anti-Tau (Sigma, #T9450, 1:100) and Anti-mouse-Alexa Flour 488 (Abcam, #ab150105, 1:500).

Dravid A., Raos B., Svirskis D, & O’Carroll S.J. (2021). Optimised techniques for high-throughput screening of differentiated SH-SY5Y cells and application for neurite outgrowth assays. Scientific Reports, 11, 23935.

+ Open protocol

+ Expand

Immunocytochemistry Staining for Neural Cell Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

For O4 staining, the primary antibodies mouse anti-O4 (Sommer and Schachner 1981 (link)) (1:30) were applied on the living cells in KRHA (Krebs–Ringer-Hepes buffer with 0.1% (w/v) BSA) for 30 min, then the cells were washed with PBS and fixed with 4% (w/v) PFA in PBS for 10 min at RT.
The staining against other epitopes was performed on fixed cells. The cells were blocked with 10% (w/v) NGS, 0.1% (v/v) Triton in PBS for 30 min and incubated for additional 30 min with primary antibodies in the blocking solution. The following primary antibodies have been used: rabbit anti-platelet-derived growth factor receptor α (PDGFRα) (Santa Cruz Biotechnology) 1:200, mouse anti-myelin basic protein (MBP) (Millipore) 1:50, mouse anti-βIII-tubulin (Sigma-Aldrich) 1:300, rabbit anti-glial fibrillary acidic protein (GFAP) (Dako, Hamburg, Germany) 1:400, mouse anti-Nestin (Millipore) 1:300, mouse anti-bromodeoxyuridine BrdU (Roche) 1:50. Further on, the cells were washed 3×10 min with PBS, incubated with secondary antibodies (see immunohistochemistry) and 4′,6-diamidino-2-phenylindole DAPI 1:50,000 for 30 min, washed again 3×10 min with PBS and mounted in PBS/glycerin 1:1.

Safina D., Schlitt F., Romeo R., Pflanzner T., Pietrzik C.U., Narayanaswami V., Edenhofer F, & Faissner A. (2016). Low-density lipoprotein receptor-related protein 1 (LRP1) is a novel modulator of radial glia stem cell proliferation, survival and differentiation. Glia, 64(8), 1363-1380.

+ Open protocol

+ Expand

Immunocytochemistry of Neurospheres and Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

For immunocytochemistry, samples (neurospheres or neurons: derived from the 201B7 line) were plated onto poly-L-ornithine/fibronectin-coated chamber slide glasses (Iwaki) and fixed in 4% PFA/PBS for 30 min at room temperature. The slides were rinsed with PBS three times and permeabilized with 0.3% Triton X-100/PBS for 5 min at room temperature. After blocking with Blocking One (Nacalai Tesque, 03953-95) for 15 min at room temperature, the slides were incubated at 4°C overnight with the following antibodies: rabbit anti-α-tubulin (Cell Signaling Technology, 2144; 1:500), mouse anti-β-III tubulin (Sigma-Aldrich, T8660-2ML; 1:500), rabbit anti-tau (Dako, A0024; 1:500), rabbit anti-p38 MAPK (Cell Signaling Technology, 8690; 1:500), rabbit anti-phospho-p38 MAPK (Cell Signaling Technology, 4511; 1:500), rabbit anti-phospho-CDC25B (Thermo Fisher Scientific, PA5-104568; 1:500), and rabbit anti-acetyl-α-tubulin (Lys40) (Cell Signaling Technology, 5335; 1:500). After washing three times with PBS, the samples were incubated with secondary antibodies conjugated to Alexa 488 (Thermo Fisher Scientific, A-11034) or Alexa 555 (Thermo Fisher Scientific, A-21424; 1:500) for 60 min at room temperature and then subjected to nuclear counterstaining with Hoechst 33258 (Sigma-Aldrich, B2883; 10 μg/mL). The samples were analyzed with an all-in-one fluorescence microscope (BZ-700 or BZ-800, Keyence).

Kase Y., Sato T., Okano Y, & Okano H. (2022). The GADD45G/p38 MAPK/CDC25B signaling pathway enhances neurite outgrowth by promoting microtubule polymerization. iScience, 25(4), 104089.

+ Open protocol

+ Expand

Immunostaining of Cortical Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cortical neurons were fixed with 4% paraformaldehyde and 4% sucrose at RT for 20 min, washed three times with PBS for 5 min each, permeabilized in 0.15% Triton X-100 for 15 min, and then incubated in blocking buffer (4% BSA and 3% normal goat serum) in PBS for 1 hr. Fixed neurons were next incubated with primary antibodies diluted in blocking buffer at 4°C overnight. Primary antibodies were as follows: rabbit anti-TOM20 (1:100, Santa Cruz), anti-myo6 (1:100, Sigma-Aldrich), anti-synaptophysin (1:400, Santa Cruz), and anti-SNPH (1:250); mouse anti-βIII-tubulin (1:5000, Sigma-Aldrich), anti-MAP2 (1:5000, BD Biosciences), anti-SV2 (1:2000, DSHB), anti-myo6 (1:50, Santa Cruz), and anti-cyto c (1:100, BD Biosciences). After washing three times with PBS at RT for 10 min each, samples were incubated with secondary fluorescent antibodies (Alexa 488, 546, 594 or 633 conjugated, Thermo Fisher Scientific) diluted in blocking buffer for 60 min, rewashed with PBS, and mounted with Fluoro-Gel mounting medium (Electron Microscopy Sciences) before imaging.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!