OSCs and differentiated OSCs were fixed in 4% paraformaldehyde (PFA, Wako Pure Chemical Industries, Ltd., Osaka, Japan) for 10 minutes and rinsed 3 times with PBS. Nonspecific binding was blocked with 5% goat serum in PBS for 20 minutes, and spheres and cells were incubated overnight at 4°C with primary antibodies diluted in PBS containing 2% goat serum. The primary antibodies used in this study include mouse anti‐Nestin antibody (Abcam Plc., Cambridge, MA), mouse anti‐Musashi‐1 antibody (Abcam Plc.), mouse anti‐βIII‐tubulin antibody (Abcam Plc.), rabbit antigalactocerebroside (GalC) antibody (Millipore Corp., Billerica, MA), mouse antiglial fibrillary acidic protein (GFAP) antibody (Abcam Plc.), and mouse antiolfactory marker protein (OMP) antibody (Abcam Plc.). Alexa Fluor 488‐ or Alexa Fluor 546‐conjugated goat antirabbit IgG or antimouse IgG antibodies (Life Technologies Corp.) were used as the secondary antibodies. Nuclei were stained with Hoechst 33258 (Dojindo Molecular Technologies, Inc., Kumamoto, Japan) 15 . Images were obtained by confocal laser scanning microscopy (Leica Microsystems GmbH, Wetzlar, Germany). Antibody specificity was confirmed using a universal negative control immunoglobulin (Dako A/S, Glostrup, Denmark).
Mouse anti βiii tubulin antibody
Manufactured by Abcam
Sourced in United Kingdom
Mouse anti‐βIII‐tubulin antibody is a primary antibody that specifically binds to the βIII isoform of tubulin. Tubulin is a structural protein that is a key component of microtubules, which are essential for various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Confocal laser scanning microscopy, by Leica (1 mentions)
Mouse anti nestin antibody, by Abcam (1 mentions)
Hoechst 33258, by Dojindo Laboratories (1 mentions)
Alexa fluor 488 anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 350 anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Vectashield medium, by Vector Laboratories (1 mentions)
2 protocols using mouse anti βiii tubulin antibody
1
Immunofluorescence Analysis of Stem Cells
2
Neurons Expressing GABA Receptors
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Immunostainings were performed on tissues from 4 animals per group. Double immunofluorescent stainings for GABAB2 receptor and βIII tubulin (a marker of all neuronal cells) were performed to determine the percentage of neurons expressing GABAB2 receptors. Sections were incubated overnight at room temperature with a rabbit anti-GABAB2 receptor antibody (1∶300, ref. 52248, Abcam, Cambridge, UK) and mouse anti-βIII Tubulin antibody (1∶2000, ref. G7121, Promega, San Luis Obispo, CA USA); antibodies were diluted in PBS-T-azide (PBS 0.02 M containing 0.3% Triton X-100 and 0.02% sodium azide). Following several washes in PBS, sections were incubated for 2 h at room temperature in Alexa Fluor® 488 anti-rabbit IgG and Alexa Fluor® 350 anti-mouse IgG (Invitrogen, California, USA), diluted 1∶1000 in PBS-T. Following several washes in PBS, sections were mounted in Vectashield medium (Vector Laboratories, Burlingame, USA). A similar procedure was performed for GABAB2 receptor-GABAB1 receptor double immunostaining, using the anti-GABAB1 receptor antibody (1∶500; mouse anti-GABAB1 receptor, ref. 55051, Abcam, Cambridge, UK) and Alexa Fluor® 546 anti-mouse IgG.
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