Embryonic mice were decapitated and heads were fixed in 4% paraformaldehyde (PFA) in PBS at 4°C overnight. Postnatal animals were perfused and brains were dissected out, then fixed in 4% PFA in PBS at 4°C overnight. Parasagittal sections 7μm thick were prepared from fixed tissues embedded in paraffin. Hematoxylin and eosin (H&E), immunofluorescence, and immunohistochemical staining of tissue sections were performed as previously described (Park et al., 2016a (link); Park et al., 2016b (link)). Antibodies used were Yap (1:500; abcam ab56701), BLBP (1:200; Millipore ABN14), Calbindin (1:200; Sigma, C9848), S100β (1:200; Novus Biologicals, NB110–57478), GFAP (1:200; Thermo Scientific, RB-087), BrdU (1:500; abcam, ab6326), PCNA (1:250; Proteintech, 10205–2-P), pH3 (1:500; Millipore Sigma, 06–670), NeuN (1:250; Millipore, MAB377), Pax6 (1:200; Covance, RBP-278), Sox9 (1:200; Millipore, AB5535), Pals1 (1:200; Proteintech), Pan-Crb (1:200; (Cho et al., 2012 (link))), NMIIB (1:500; Covance, PRB-445P), N-Cadherin (1:500; BD, 610920), and β-Catenin (1:500; BD, 610153). Species-specific secondary antibodies conjugated to Alexa Fluor 488 (1:250; Invitrogen) or Cy3 (1:250; Jackson Immunochemical) were used for immunofluorescence. Nuclei were stained with Hoechst 22358 (1:500; Invitrogen).
Ab6326
Manufactured by Proteintech
Ab6326 is an antibody product offered by Proteintech. It is a laboratory tool used for scientific research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Calbindin, by Merck Group (1 mentions)
Ab5535, by Proteintech (1 mentions)
Prb 445p, by BD (1 mentions)
Hoechst 22358, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Mab377, by Fortrea (1 mentions)
β catenin, by BD (1 mentions)
N cadherin, by BD (1 mentions)
Ab109520, by Abcam (1 mentions)
In cell 2200 automated 991 microscope, by GE Healthcare (1 mentions)
In cell 2200 developer software version 1, by GE Healthcare (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
2 protocols using ab6326
1
Immunohistochemical Analysis of Embryonic and Postnatal Mouse Brains
2
High-Content Immunofluorescence Assay
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Cells were grown in a 96-well plate and fixed with 4% paraformaldehyde for 10 min at RT. Cells were then washed twice with PBS and permeabilized by incubating with 0.4% Triton X-100 in PBS for 10 min at RT. After a PBS wash, cells were blocked 30 min at RT using 1% BSA in PBS supplemented with 0.1% Tween-20 (PBST). Primary antibodies (details found at the end of the section) were diluted in 1% BSA-PBST and incubated overnight. For BrdU staining, cells were treated with DNaseI and MgCl2 simultaneously with the primary antibody. Cells were then washed with PBS and incubated with their respective secondary antibody for 1 h at RT. Nuclei were stained with DAPI (Sigma-Aldrich). Images were acquired using INCell 2200 automated 991 microscope (GE) and INCell 2200 Developer software version 1.8 (GE) was used for image analysis. Antibodies used in this study are: p21CIP (Abcam, Cat# ab109520), Ki67 (Abcam, Cat# ab92742), BrdU (Abcam, ab6326; 1:500) and F9 (Proteintech, Cat# 21481–1-AP).
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