For immunostaining, cells were fixed with 4% paraformaldehyde, blocked and incubated with primary antibodies overnight at 4 °C. They wer e then stained with Alexa Fluor conjugated secondary antibodies Goat anti-Rabbit IgG (H+L) (Thermo Fisher) and Goat anti-Mouse IgG (H+L) (Thermo Fisher) at RT for one hour. Nuclear staining was performed with DAPI (BD Bioscience). The following primary antibodies were used in this study: NANOG (D73G4) XP® Rabbit mAb #4903 (1:300) (4903S, Cell Signaling), NESTIN (1:200) (10C2, Biolegend 656801), EDC4 (1:50) (Abcam ab72408) DDX6 (Novus NB200–192), βIII-tubulin (TUJ1, Biolegend 801211), SOX1 (R&D SYSTEMS AF3369), SOX2 (R&D SYSTEMS AF2018), MYOSIN (MF-20 Hybridoma bank), LSM14A (1:50) (N3C3, GeneTex GTX120902), LSM14B (1:50) (Sigma Aldrich HPA061189).
βiii tubulin tuj1
Manufactured by BioLegend
βIII-tubulin (TUJ1) is a specific marker for post-mitotic neurons. It is a member of the tubulin family of proteins, which are the structural components of microtubules. βIII-tubulin is predominantly expressed in neurons and is essential for the assembly and stabilization of the neuronal cytoskeleton.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti mouse igg h l, by Thermo Fisher Scientific (1 mentions)
Goat anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by BD (1 mentions)
Brachyury, by R&D Systems (1 mentions)
Iba 1, by Abcam (1 mentions)
Ang 2, by Abcam (1 mentions)
Cxcl10, by Thermo Fisher Scientific (1 mentions)
Brain derived neurotrophic factor (bdnf), by Thermo Fisher Scientific (1 mentions)
Erbb4, by Abcam (1 mentions)
Glial fibrillary acidic protein (gfap), by Abcam (1 mentions)
Alexa fluor 647, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Vectashield mounting medium, by Vector Laboratories (1 mentions)
Dapi h 1200, by Vector Laboratories (1 mentions)
Alexa fluor conjugated secondary antibodies, by Merck Group (1 mentions)
Hoechst, by Merck Group (1 mentions)
C fos, by Cell Signaling Technology (1 mentions)
3 protocols using βiii tubulin tuj1
1
Immunostaining Protocol for Stem Cell Markers
2
Histological Analysis of Cortical Organoids
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Brain cortical organoids at 20-100 days in culture were fixed with 10% formalin, dehydrated with ethanol, and embedded in paraffin blocks. 10 μm thick sections were deparaffinized with xylene, rehydrated in a graded series of ethanol and double distilled water in a standard manner, and then stained with H&E. To detect protein expression, the sections were heated to unmask the epitopes, blocked with 10% goat serum, then primary antibodies were added overnight [Sox2 from EMD Millipore, β III tubulin (TUJ1, BioLegend, San Diego, CA)]; IBA-1, NRG1, ErbB4, GFAP, CXCR3, Ang2 and Ang1 from Abcam; Brachyury from R&D Systems; Cleaved caspase 3 (CC3), NFL and NFM from Cell Signaling, Danvers, MA; BDNF from Thermo Fisher Scientific and CXCL-10 from PeproTech (Rocky Hill, NJ). Fluorescence staining was performed using fluorochrome-labelled secondary antibodies (Alexa Fluor 488, Alexa Fluor 647, Invitrogen). The sections were covered with Vectashield mounting medium containing DAPI (H-1200) obtained from Vector Laboratories Inc (Burlingame, CA).
3
Immunocytochemistry Protocol for Cell Labeling
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For immunocytochemistry (ICC), cells were fixed with 4% paraformaldehyde (PFA) 15 min on ice, washed and incubated in blocking solution (1% Bovine serum albumin (BSA), 0,25% Triton X-100 (0,25% Tx) in 0.1M PBS) for 1 h at RT. Primary antibody (βIII tubulin (Tuj1, 1:1000, BioLegend), c-Fos (1:200, Cell Signaling), ChR2 (1:500, Progen), mCherry (1:500, Abcam) GFP (1:500, ThermoFisher Scientific) was added to the cells in blocking solution and incubated O/N at 4 °C. After washing, the cells were incubated 1 h at RT with Alexa-Fluor-conjugated secondary antibodies (568 goat and 488 goat and donkey) and Hoechst (Sigma Aldrich).
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