Brain organoids were fixed in 4% paraformaldehyde for 45 min. After two washes in PBS, organoids were moved to 30% sucrose solution overnight at 4 °C. Organoids were then embedded in optimal cutting temperature (OCT) compound and flash frozen at –80 °C. Sections (20 µm) were cut with a cryostat (HM560 Microm Microtech). After permeabilization with 0.3% Triton and blocking with 2% goat serum (Gibco, 16210064) and 1% bovine serum albumin (BSA), sections were incubated for 1 hr with the following primary antibodies: SOX2 1:100 (Abcam, ab93689), PAX6 1:100 (Sigma-Aldrich, AMAb91372), βIII-tubulin 1:500 (Sigma-Aldrich, T8578), TBR1 1:100 (Abcam, ab31940), PSD95 1:500 (Sigma-Aldrich, MABN68), NeuN 1:100 (Sigma-Aldrich, MAB377), MAP2 1:100 (Sigma-Aldrich, ZRB2290), GFAP 1:200 (Sigma-Aldrich, G3893). After PBS washes, DAPI (100–1000 ng/ml) and secondary antibodies labeled with Alexa Fluor 488 or 594 1:500 (Invitrogen, #A-11037, #A-11029) were incubated for 1 hr. Slides were mounted using Fluoromount Aqueous Mounting Medium (Sigma Aldrich). Images were acquired using a microscope Axio Observer Z1 (Carl Zeiss), Objective 10 X, and analyzed using Fiji software.
T8578
Manufactured by Abcam
T8578 is a laboratory product manufactured by Abcam. It is a laboratory equipment item, but a detailed description is not available while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Axio observer z1, by Zeiss (6 mentions)
Ab93689, by Merck Group (2 mentions)
Ab31940, by Merck Group (2 mentions)
Fluoromount aqueous mounting medium, by Merck Group (2 mentions)
Amab91372, by Merck Group (2 mentions)
Mabn68, by Merck Group (2 mentions)
Mab377, by Merck Group (2 mentions)
Zrb2290, by Merck Group (2 mentions)
G3893, by Merck Group (2 mentions)
Hm560, by Microtech (2 mentions)
2 protocols using t8578
1
Immunohistochemical Analysis of Brain Organoids
2
Immunohistochemical Analysis of Brain Organoids
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Brain organoids were fixed in 4% paraformaldehyde for 45 min. After two washes in PBS, organoids were moved to 30% sucrose solution overnight at 4 °C. Organoids were then embedded in optimal cutting temperature (OCT) compound and flash frozen at –80 °C. Sections (20 µm) were cut with a cryostat (HM560 Microm Microtech). After permeabilization with 0.3% Triton and blocking with 2% goat serum (Gibco, 16210064) and 1% bovine serum albumin (BSA), sections were incubated for 1 hr with the following primary antibodies: SOX2 1:100 (Abcam, ab93689), PAX6 1:100 (Sigma-Aldrich, AMAb91372), βIII-tubulin 1:500 (Sigma-Aldrich, T8578), TBR1 1:100 (Abcam, ab31940), PSD95 1:500 (Sigma-Aldrich, MABN68), NeuN 1:100 (Sigma-Aldrich, MAB377), MAP2 1:100 (Sigma-Aldrich, ZRB2290), GFAP 1:200 (Sigma-Aldrich, G3893). After PBS washes, DAPI (100–1000 ng/ml) and secondary antibodies labeled with Alexa Fluor 488 or 594 1:500 (Invitrogen, #A-11037, #A-11029) were incubated for 1 hr. Slides were mounted using Fluoromount Aqueous Mounting Medium (Sigma Aldrich). Images were acquired using a microscope Axio Observer Z1 (Carl Zeiss), Objective 10 X, and analyzed using Fiji software.
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