Slides were thawed to room temperature, and lines drawn to partition the edges of sections on the glass slides using a PAPpen (ImmEdge Pen; Vector Labs H4000). The sections were rehydrated, blocked, permeabilized and immunostained with primary antibodies: PAX6 (BioLegend, 1:200), SOX2 (Santa Cruz, 1:100), β-tubulin III (Sigma, 1:1000), FOXG1 (Takara, 1:500), SOX10 (Santa Cruz, 1:100), Ki67 (Thermo Fisher, 1:200), Nestin (Millipore, 1:200), GFAP (Millipore, 1:400), S100beta (DAKO Potts, 1:400), CTIP2 (Abcam, 1:500), SATB2 (Abcam, 1:50), MAP2AB (Abcam, 1:1000-2000), NeuN (Millipore, 1:200), TBR1(Abcam, 1:500), BRN2 (Millipore, 1:500), MEF2C (Novis, 1:200), AT8 (Invitrogen, 1:1000), PHF1 (gift from Peter Davies,1:1000), Total Tau (DAKO Potts, 1:200), DA9 (gift from Peter Davies, 1:1000), ELAVL4 (Santa Cruz, 1:200), TIA1 (Santa Cruz, 1:100), G3BP1 (Protein Tech, 1:200), VGLUT1 (gift from Susan Morton, Tom Jessell, 1:16,000), Calbindin1 (Swant, 1:10,000), Homer1 (SYSY, 1:250) and Synapsin1 (Millipore, 1:100). Primary antibodies were incubated overnight at 4C, washed three times with PBS and then incubated with corresponding Alexa Fluor conjugated secondary antibody (1:333-1000) for 1 hour at room temperature. Sections were coverslipped and imaged using fluorescence and confocal microscopy (Zeiss AXIO Observer.Z1; Zeiss 780).
Foxg1
Manufactured by Takara Bio
FOXG1 is a laboratory DNA/RNA detection reagent. It is used to detect the presence and expression levels of the FOXG1 gene in biological samples. FOXG1 is a transcription factor that plays a critical role in the development and function of the central nervous system.
Automatically generated - may contain errors
Lab products found in correlation
Ctip2, by Abcam (3 mentions)
Axio observer z1, by Zeiss (1 mentions)
β tubulin 3, by Merck Group (1 mentions)
Glial fibrillary acidic protein (gfap), by Merck Group (1 mentions)
Neuronal nuclei (neun), by Merck Group (1 mentions)
Nestin, by Merck Group (1 mentions)
Synapsin 1, by Merck Group (1 mentions)
G3bp1, by Proteintech (1 mentions)
Map2 ab, by Abcam (1 mentions)
Elavl4, by Santa Cruz Biotechnology (1 mentions)
Satb2, by Abcam (1 mentions)
Tia 1, by Santa Cruz Biotechnology (1 mentions)
Sox10, by Santa Cruz Biotechnology (1 mentions)
Immedge pen, by Vector Laboratories (1 mentions)
Fv1200, by Olympus (1 mentions)
Cryostat, by Leica (1 mentions)
Target retrieval solution, by Agilent Technologies (1 mentions)
Rhodopsin, by Merck Group (1 mentions)
Calbindin, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Nacalai Tesque (1 mentions)
3 protocols using foxg1
1
Immunohistochemical Characterization of Neural Tissue
2
Immunohistochemical Analysis of Retinal Cell Markers
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Immunohistochemistry was performed as described6 (link). Aggregates were fixed with 4% PFA and sectioned with a cryostat (Leica). Frozen sections were treated with or without heat-based antigen retrieval in Target Retrieval solution (Dako) at 105 °C for 15 min. The primary antibodies used in this study were as follows: Chx10 (sheep; 1:500; Exalpha), Rx (guinea pig; 1:2000; Takara), Pax6 (mouse; 1:1000; BD Biosciences), Mitf (mouse; 1:500; Exalpha), Crx (rabbit; 1:200; Takara), Recoverin (rabbit; 1:1000; Proteintech), NRL (goat; 1:500; R&D Systems), RXR-gamma (rabbit; 1:500; Spring Bioscience), Rhodopsin (mouse; 1:1000; Sigma; RET-P1), S-opsin (rabbit; 1:500; Millipore), L/M-opsin (rabbit; 1:500; Millipore), PKCalpha (rabbit; 1:500; Sigma), HuNu (Ms; 1:1000; Millipore), Calbindin (rabbit; 1:500; Millipore), N-cadherin (mouse; 1:500; BD Biosciences), FoxG1 (rabbit; 1:2000; Takara), Ctip2 (rat; 1:500; abcam), Tbr1 (rabbit; 1:500; abcam). Nuclear counter-staining was performed with DAPI (Nacalai). Stained sections were analyzed with a fluorescence microscope (BIOREVO; Keyence) and a confocal microscope (FV1200; Olympus). Image analyses were performed with ImageJ 1.48 v software (NIH).
3
Immunohistochemical Analysis of Organoid Cultures
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Organoids were randomly selected and fixed in 4% PFA in PBS for 2–4 hours. The organoids were then cryopreserved in 25% sucrose overnight, embedded in O.C.T. (Sakura), and frozen on dry ice before being stored at −80°C. Serial cryosections were obtained with a thickness of 12–16 µm. Immunostaining was performed by incubating the sections in blocking solution (PBS, 10% Donkey Serum, 1% Triton-100) for 1 hour, followed by incubation with primary antibodies (overnight, 4°C) and secondary antibodies (1–2 hours, from Jackson ImmunoResearch or ThermoFisher Scientific). The slides were then mounted with coverslips using VECTASHIELD (Vector Labs) and imaged on a Zeiss microscope equipped with an apotome module and ZEN 3.3 (ZEN pro) software. Three cell lines were used for immunocytochemical analyses, and a minimum of four organoids per line were analyzed. Images were acquired randomly to cover the entire extent of the organoid. Antibody list: FOXG1 (rabbit, 1:200, Takara) and PAX6 (mouse, 1:200, BD Bioscience), EOMES (rabbit, 1:1000, Abcam), FOXP2 (goat, 1:200, Santa Cruz), GAD1 (mouse, 1:200, Chemicon), HuC/D (mouse, 1:200, Invitrogen), SOX1 (goat, 1:100, R&D Systems), CTIP2 (rat, 1:500, Abcam).
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