α-Vax1 and α-Vax2 were produced as reported previously (Mui et al., 2005 (link)). Commercially available antibodies against the following proteins were used: mouse anti-Myc (Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-GFP (Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-tubulin β-III (Tuj1; Covance, Princeton, NJ, USA), goat anti-Sox2 (Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-Nestin (RC2; Millipore, Billerica, MA, USA), mouse anti-β-galactosidase (Developmental Studies Hybridoma Bank, DSHB), mouse anti-NF160 (Developmental Studies Hybridoma Bank [DSHB], Iowa City, IA, USA), goat anti-Sdc2 (Santa Cruz Biotechnology, Dallas, TX, USA), goat anti-Sdc3 (Santa Cruz Biotechnology (Dallas, TX, USA), for immunohistochemistry), rabbit anti-Sdc3 (Abcam (UK), for Western blot), rabbit anti-Glp1 (Santa Cruz Biotechnology, Dallas, TX, USA), and rabbit anti-Pax2 (Invitrogen, Carlsbad, CA, USA) antibodies.
Mouse anti β 3 tubulin tuj1
Manufactured by Fortrea
Sourced in United States
Mouse anti-β-III tubulin (Tuj1) is a monoclonal antibody that specifically recognizes the β-III isoform of tubulin, a key structural protein found in neurons. This antibody is commonly used as a marker for neuronal cells and their differentiation in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Cy3 or fitc conjugated secondary antibody, by Jackson ImmunoResearch (2 mentions)
Rat anti mbp, by Abcam (2 mentions)
Anti rabbit cy5, by Jackson ImmunoResearch (2 mentions)
Anti rat fitc, by Jackson ImmunoResearch (2 mentions)
Anti mouse cy3, by Jackson ImmunoResearch (2 mentions)
Mouse anti nestin, by BD (2 mentions)
Rabbit anti gr, by Thermo Fisher Scientific (2 mentions)
Mouse anti myc, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti pax2, by Thermo Fisher Scientific (1 mentions)
Goat anti sox2, by Santa Cruz Biotechnology (1 mentions)
Mouse anti β galactosidase, by Developmental Studies Hybridoma Bank (1 mentions)
Mouse anti gfp, by Santa Cruz Biotechnology (1 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Sheep anti mouse igg cy3, by Merck Group (1 mentions)
Mouse anti neun, by Merck Group (1 mentions)
Rabbit anti cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Mouse anti brdu, by BD (1 mentions)
Mouse anti brn3a, by Merck Group (1 mentions)
710 confocal microscope, by Zeiss (1 mentions)
5 protocols using mouse anti β 3 tubulin tuj1
1
Antibody Validation for Protein Detection
2
Immunofluorescence Staining of Neural Stem Cells
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aNSCs were fixed with 4 % PFA and permeabilized with 0.7 % Triton diluted in DPBS, no calcium, no magnesium (PBS, Thermo Fisher Scientific) for 15 min. Samples were then incubated with primary antibodies (resuspended in PBS supplemented with 1 % BSA, 5 % donkey serum and 0.1 % Triton) under 4 °C overnight. After being washed with PBS, the samples were then further incubated with secondary antibodies (resuspended in PBS supplemented with 7 % BSA and 1:1000 DAPI) for 1 h. The samples were then conserved by ProLong™ Gold Antifade reagent (Thermo Fisher Scientific). Following antibody/reagent were used for immunofluorescence staining: mouse anti-Tubulin βIII (Tuj1, 1:400, Covance, Princeton, United States), sheep anti-mouse IgG Cy3 (1:400, Sigma-Aldrich) and DAPI (1:3000). The coverslips were then imaged by the ZEISS Apotome 3 (Carl Zeiss AG, Jena, Germany) under the 20x or 40x objectives. The neuron population was then scored with ImageJ software and numerated manually.
3
Immunostaining of Neural Stem Cells
4
Immunostaining of Neural Stem Cells
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NSCs were fixed in 4% paraformaldehyde (PFA) and immunostained with rabbit anti-GR (1:500, Thermo Scientific, Rockford IL), mouse anti-Nestin (1:500, BD Biosciences, San Jose CA), and rat anti-BrdU (1:500) overnight at 4°C, and then treated with Cy3 anti-mouse, FITC anti-rat, and Cy5 anti-rabbit (1:500, Jackson ImmunoResearch, West Grove PA) for 2h.
For differentiation studies, cells were immunostained (overnight, 4°C) with mouse anti-β-III tubulin (Tuj1; 1:1000, Covance, Princeton NJ) or rat anti-MBP (1:100, Abcam, Cambridge MA) followed by 2h incubation with Cy3 or FITC-conjugated secondary antibodies (1:500, Jackson). The nuclear stain DAPI was added prior to mounting coverslips in 1,4-diazabicyclo[2.2.2]octane (DABCO). Images were obtained with an inverted fluorescence microscope (20× objective; Zeiss, Oberkochen, Germany), and blind cell counts were taken using Metamorph software. 20 random visual fields were analyzed per coverslip. Data are the percentage of positive cells in relation to the total number of DAPI-stained nuclei present in the culture. All experiments were independently replicated at least 3 times.
For differentiation studies, cells were immunostained (overnight, 4°C) with mouse anti-β-III tubulin (Tuj1; 1:1000, Covance, Princeton NJ) or rat anti-MBP (1:100, Abcam, Cambridge MA) followed by 2h incubation with Cy3 or FITC-conjugated secondary antibodies (1:500, Jackson). The nuclear stain DAPI was added prior to mounting coverslips in 1,4-diazabicyclo[2.2.2]octane (DABCO). Images were obtained with an inverted fluorescence microscope (20× objective; Zeiss, Oberkochen, Germany), and blind cell counts were taken using Metamorph software. 20 random visual fields were analyzed per coverslip. Data are the percentage of positive cells in relation to the total number of DAPI-stained nuclei present in the culture. All experiments were independently replicated at least 3 times.
5
Embryonic Tissue Immunolabeling Protocol
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