Rat monoclonal ATE1 antibody (EMD Millipore, MABS436) was previously described in Wang et al. (2011) (link). Rabbit polyclonal antibody to arginylated actin (R-actin) was obtained from EMD Millipore (ABT264) as part of their product validation study. Batch 13 of the antibody, not currently licensed for commercial use, is shown in the majority of the study. Anti-Doublecortin antibody (ab18723 dilutions: 1:200 for immunofluorescence) was purchased from Abcam. Mouse monoclonal anti- β-III tubulin was from R & D Systems (MAB1195, dilutions: 1:100 for immunofluorescence). Rhodamine-phalloidin was purchased from Sigma and used at the working concentration of 100 nM.
Anti β 3 tubulin
Manufactured by R&D Systems
Sourced in United States
Anti-β-III tubulin is a monoclonal antibody that recognizes the β-III isoform of the tubulin protein. 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 Anti-β-III tubulin antibody can be used to detect and quantify the expression of the β-III tubulin isoform in various cell and tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
Abt264, by Merck Group (2 mentions)
Rhodamine phalloidin, by Merck Group (2 mentions)
Mabs436, by Merck Group (2 mentions)
R actin, by Merck Group (2 mentions)
Anti doublecortin antibody, by Abcam (2 mentions)
Fitc conjugated pna, by Merck Group (2 mentions)
Cd44 pe, by BD (1 mentions)
Cd45 pe, by BD (1 mentions)
Cd29 pe, by BD (1 mentions)
Cd90 fitc, by BD (1 mentions)
Cd105 pe, by BD (1 mentions)
Cd34 fitc, by BD (1 mentions)
Cd31 apc, by BD (1 mentions)
Anti p63, by Abcam (1 mentions)
Anti nestin, by R&D Systems (1 mentions)
Anti fibronectin, by R&D Systems (1 mentions)
Anti gfap, by R&D Systems (1 mentions)
Anti vimentin, by R&D Systems (1 mentions)
Fitc anti rabbit, by Vector Laboratories (1 mentions)
Anti filaggrin, by Abcam (1 mentions)
6 protocols using anti β 3 tubulin
1
Immunofluorescence Antibody Validation
2
Immunophenotyping of Neural Stem Cells
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The primary antibodies used were: anti-filaggrin (1:500; Abcam, catalog number # ab24584), anti-fibronectin (1:200; R&D Systems, catalog number # AF19181), anti-GFAP (1:400; R&D Systems, catalog number # AF2594), anti-βIII tubulin (1:200; R&D Systems, catalog number # MAB1195), anti-vimentin (1:200; R&D Systems, catalog number # MAB2105), anti-nestin (1:200; R&D Systems, catalog number # MAB2736), anti-CNPase (1:400; Chemicon, catalog number # MAB326), CD34-FITC (Pharmingen, San Jose, CA catalog number # 553733) and anti-p63 (1:500, AbCAM, catalog number # ab53039). The secondary antibodies were: Texas Red anti-sheep (1:1,000 catalog number # TI-6000), FITC anti-sheep (1:500 catalog number # F7634), FITC anti-rabbit (1:1,000 catalog number # FI-1000), anti-sheep HRP (1:1,000 catalog number # A16-147), anti-rabbit HRP (1:1,000 catalog number # PI-1000) all from Vector Laboratories, USA. The fluorophore-conjugated antibodies for flow cytometry were: CD34-PE catalog number # 551387, CD31-APC catalog number #561814, CD45-PE catalog number # 553081, CD44-PE catalog number # 561860, CD90-FITC catalog number # 554894, CD29-PE catalog number #562801, CD105-PE catalog number # 562759 diluted 1:500, all from BD Biosciences (San Jose, CA, USA).
3
Immunofluorescence Antibody Validation
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Rat monoclonal ATE1 antibody (EMD Millipore, MABS436) was previously described in Wang et al. (2011) (link). Rabbit polyclonal antibody to arginylated actin (R-actin) was obtained from EMD Millipore (ABT264) as part of their product validation study. Batch 13 of the antibody, not currently licensed for commercial use, is shown in the majority of the study. Anti-Doublecortin antibody (ab18723 dilutions: 1:200 for immunofluorescence) was purchased from Abcam. Mouse monoclonal anti- β-III tubulin was from R & D Systems (MAB1195, dilutions: 1:100 for immunofluorescence). Rhodamine-phalloidin was purchased from Sigma and used at the working concentration of 100 nM.
4
Fluorescent Labeling of SMG Tissues
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The cultured SMG tissues were fixed with 4% paraformaldehyde (PFA) and incubated with fluorescein isothiocyanate (FITC)-conjugated PNA (1:200; Sigma–Aldrich, MO, USA). The fixed tissues were treated with anti-β III-tubulin (1:1000; R&D Systems, MN, USA), Antibody binding was detected with Alexa Fluor-conjugated secondary antibodies (1:200; Life Technologies, NY, USA), and the samples were imaged by confocal microscopy (C1; Nikon).
5
Immunofluorescence and Western Blot Analysis of SMGs
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The cultured SMGs were fixed with 4% paraformaldehyde (PFA) and incubated with fluorescein isothiocyanate (FITC)-conjugated PNA (1:200, Sigma-Aldrich, MO), anti-βIII-tubulin (1:1000, R&D Systems), anti-FGF7 (1:1000, AB Biotech), anti-FGF10 (1:2000, Millipore), or anti-ki-67 (1:1500, Abcam, UK). The antibodies were detected using Alexa Fluor 488 or 568 conjugated secondary antibodies (Life Technologies). Images were obtained using confocal microscopy (C1, Nikon). The total amount of protein obtained from the cultured tissue was calculated using a BCA protein assay kit (Pierce Biotechnology, IL) and then the following antibodies were used in Western blot analysis: anti-FGF7 (1:1000) and anti-FGF10 (1:2000). Goat anti-rabbit horseradish peroxidase conjugated secondary antibody (1:8000, Santa Cruz Biotec, CA) was used as a secondary antibody. ß-Actin antibody (1:5000, Abcam) was used as a loading control. Western blot data were quantified by comparing the relative density of target bands to the loading control by using Image J software. Additional information is supplied in the Supplementary data.
6
Immunostaining of Cell and Tissue Samples
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Cultured cells and frozen tissue sections were fixed with 4% paraformaldehyde and then stained with the following primary antibodies: anti-Sox10 (N-20; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-PDGF receptor α-chain (558774; BD Pharmingen, San Diego, CA, USA), anti-PDGF receptor β-chain (#3169; Cell Signaling Technology, Danvers, MA, USA), anti-GFP (ab13970; Abcam, Cambridge, UK), anti-Ap2α, anti-AP-2β (Cell Signaling Technology), anti-Nanog (AB5731; Millipore, Temecula, CA, USA) anti-Nestin (G-20; Santa Cruz Biotechnology), anti-Oct3/4 (MAB1759; R&D Systems, Minneapolis, MN, USA), anti-PAR4 (3G9H7; Santa Cruz Biotechnology), anti-Sox2 (Y-17; Santa Cruz Biotechnology), anti-alpha smooth muscle actin (ab32575; Abcam), anti-βIII-tubulin (MAB1195; R&D Systems), anti-glial fibrillary acidic protein (MAB360; Chemicon International, Temecula, CA, USA) and anti-GFP (MBL, Nagoya, Japan). After washing with Tris-buffered saline, the sections were stained with Alexa Fluor-488 or Alexa Fluor-568 conjugated secondary antibodies (Invitrogen). All sections were counterstained with Hoechst 33342 (Invitrogen).
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