β3 tubulin

Manufactured by Abcam

Sourced in China

β3-tubulin is a protein that is a component of microtubules, which are cytoskeletal structures involved in various cellular processes. It plays a role in the formation and maintenance of microtubules.

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Lab products found in correlation

Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Triton x 100, by Merck Group (1 mentions) Dmi4000b fluorescence microscope, by Leica (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Solarbio (1 mentions) Paraformaldehyde (pfa), by Solarbio (1 mentions) Nestin, by Merck Group (1 mentions) Glial fibrillary acidic protein (gfap), by Abcam (1 mentions) Tcs sp5 mp confocal laser scanning microscope, by Leica (1 mentions) Ssea4, by Santa Cruz Biotechnology (1 mentions) Lsm 780, by Zeiss (1 mentions) Alexa fluor 488 or 546, by Thermo Fisher Scientific (1 mentions) Anti gapdh, by Merck Group (1 mentions) F actin, by Thermo Fisher Scientific (1 mentions) Fv1000, by Olympus (1 mentions) Caspase 3, by Merck Group (1 mentions) P creb1, by Merck Group (1 mentions) Alexa fluor 568, by Thermo Fisher Scientific (1 mentions) Rhodamine phalloidin, by Thermo Fisher Scientific (1 mentions)

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Anti-β3-Tubulin (2 citations)

6 protocols using β3 tubulin

Immunofluorescence Staining of Stem Cell Markers

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Cells were fixed with 4% PFA (Solarbio, China) for 10–15 min at room temperature, permeabilized with 0.3% Triton X-100 (Sigma) for 10–15 min at room temperature, blocked with 3% bovine serum albumin (Solarbio) for 45–60 min at room temperature, and then incubated with primary antibodies against OCT4 (1:100; Santa Cruz Biotechnology), SSEA4 (1:100; Santa Cruz Biotechnology), SOX2 (1:100; Santa Cruz Biotechnology), NESTIN (1:100; Sigma), β3-tubulin (1:100; Abcam), and GFAP (1:100; Abcam) overnight at 4 °C. And then cells were incubated with secondary antibodies: Goat anti-Rabbit IgG Alexa Fluor 488 (1:200; Invitrogen) and Goat anti-Mouse IgG Alexa Fluor 594 (1:200; Invitrogen) for 1 h at 37 °C. Wash with PBS three times before each step. Nuclei were stained with DAPI (300 nM, Invitrogen) for 15 min at room temperature. Fluorescence images were captured by Leica DMI 4000B fluorescence microscope and Leica TCS SP5 MP confocal laser scanning microscope (Leica, Germany).

Bai R., Chang Y., Saleem A., Wu F., Tian L., Zhang S., Li Y., Ma S., Dong T., Guo T., Jiang Y., You Y., Lu W.J., Jiang H.F, & Lan F. (2021). Ascorbic acid can promote the generation and expansion of neuroepithelial-like stem cells derived from hiPS/ES cells under chemically defined conditions through promoting collagen synthesis. Stem Cell Research & Therapy, 12, 48.

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Live-cell Microscopy of Neuronal Biochips

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Live-cell phase microscopy was performed on a Zeiss microscope equipped with an onstage incubator maintained at 37 °C and 5% CO₂, and the immunostained biochips were observed under the same microscope equipped with fluorescence-imaging ability.
After 3 days, the neurons on biochips (inside or outside microwells) were fixed with 4% paraformaldehyde/glucose in PBS and stained with neuron-specific antibody β3-tubulin (abcam). Alex Fluor 488 goat anti-mouse (Invitrogen) was used as the secondary antibody. Finally, slides were prepared with ProLong® gold antifade with DAPI reagent as the mounting medium (Invitrogen).

Wei L., Sweeney A.J., Sheng L., Fang Y., Kindy M.S., Xi T, & Gao B.Z. (2014). Single-Neuron Axonal Pathfinding under Geometric Guidance: Low-Dose-Methylmercury Developmental Neurotoxicity Test. Lab on a chip, 14(18), 3564-3571.

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Immunofluorescent Corneal Nerve Imaging

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At POD28, corneas were fixed with 2% paraformaldehyde for 2 hours and 1% paraformaldehyde for 30 minutes at 4°C and room temperature, respectively. Moreover, corneas were dehydrated using 20% and 10% sucrose solutions, followed by a permeation step consisting of 3 washes with PBS for 15 minutes. Samples were then blocked for 2 hours at room temperature with 5% normal donkey serum in PBS, followed by an overnight incubation with primary antibodies (β3 tubulin, 1:500; Abcam). Upon overnight incubation, the samples were washed 3 times with PBS for 30 minutes and incubated with fluorescently labeled secondary antibodies for 1 hour (1:1000 dilution). Images were captured using a confocal microscope (LSM780; Carl Zeiss Microscopy Ltd., Germany).

Yoo Y.S., Park S., Eun P., Park Y.M., Lim D.H, & Chung T.Y. (2022). Corneal Neuro-Regenerative Effect of Transcutaneous Electrical Stimulation in Rabbit Lamellar Keratectomy Model. Translational Vision Science & Technology, 11(10), 17.

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Antibody validation for neuroscience research

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The following antibodies were used: homemade anti-Myo1b polyclonal antibodies (Almeida et al., 2011 (link)), 1:1,000 for Western blot and 1:50 for immunofluorescence; mouse monoclonal antibodies against Tau-1 (1:100) purchased from EMD Millipore and β-3 tubulin (1:150) purchased from Abcam; and rabbit polyclonal anti-GAPDH (1:20,000 for Western blot) purchased from Sigma-Aldrich. We also used Alexa Fluor 488–, 546–, or 647–coupled secondary antibodies against mouse or rabbit IgG (1:400) and horseradish peroxidase–conjugated secondary antibodies against mouse or rabbit IgG (1:5,000; Invitrogen); and anti–rabbit IgG Cy3 (1:400; Jackson ImmunoResearch Laboratories, Inc.; or 1:500; Molecular Probes). Alexa Fluor 488– or 546–conjugated phalloidin was used to detect F-actin (1:400; Invitrogen).

Iuliano O., Yoshimura A., Prospéri M.T., Martin R., Knölker H.J, & Coudrier E. (2018). Myosin 1b promotes axon formation by regulating actin wave propagation and growth cone dynamics. The Journal of Cell Biology, 217(6), 2033-2046.

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Immunohistochemistry of Neural Markers

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Briefly, PBS was used to wash the slides three times, which were fixed in 4% paraformaldehyde. Ten minutes later, 0.1% Triton X-100 in PBS was employed to permeabilize cells for 15 min. Then, tissues were blocked with 2% BSA in TBST for 1h. The antibody (NeuN [1:400, cat. No. ABN78; Millipore], p-CREB1 [1:200, cat. No. ab32096; Abcam], β3-Tubulin [1:400, cat. No. MAB1637; Millipore], and caspase-3 [1:100, cat. No. sc-7272; Santa Cruz]) was incubated overnight, and the second antibody was incubated for 1h. DAPI was used to stain the nuclei. Subsequently, the images were photographed using OLYMPUS FV1000.

Wu N., Liu H., Lv X., Sun Y, & Jiang H. (2023). Neobaicalein prevents isoflurane anesthesia-induced cognitive impairment in neonatal mice via regulating CREB1. Clinics, 78, 100201.

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Wistar Rat Eye Tissue Sectioning

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All animal experiments were conducted under a protocol approved by the University Health Network (UHN) animal care committee and conformed to local, provincial, and federal regulations. Wistar rats eyes were collected and sectioned as previously described (Guo et al., 2018 (link)). Briefly, p21–p30 rats were killed by CO₂ asphyxiation. Eyes were then immediately enucleated and processed for analyses. Fixed eye tissues were dehydrated using 30% sucrose overnight before OCT embedding and cryosectioning. Tissue sections were permeabilized using 0.2% triton-X in PBS. After blocking, primary antibodies to ANXA4 (1:200; Sigma), GFAP (1:200; Sigma), β3 tubulin (1:200; Abcam) were incubated overnight. Samples were then incubated in Alexa-Fluor-488 or Alexa-Fluor-568 secondary antibodies (1:200; Life Technologies) for 1 h. Rhodamine phalloidin (1:50, Life Technologies) was incubated for 1 h, washed and coverslipped. Images were taken using a Nikon Eclips-Ti Confocal Microscope.

Vicic N., Guo X., Chan D., Flanagan J.G., Sigal I.A, & Sivak J.M. (2022). Evidence of an Annexin A4 mediated plasma membrane repair response to biomechanical strain associated with glaucoma pathogenesis. Journal of cellular physiology, 237(9), 3687-3702.

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