Cell cultures were fixed with 4% paraformaldehyde for 20 min and permeabilized with 0.3% Triton X-100 (Sigma). Primary antibodies were incubated overnight at 4°C in phosphate buffer saline (PBS) plus 10% normal goat serum. Cells were washed three times and secondary antibodies incubated for 2 h in the same solution, washed three times and incubated with Hoechst for nuclei staining. Primary antibodies were used with the following dilutions: Oct 3/4, 1:200 (BD 611202); Sox2, 1:1000 (Millipore AB5603); GFP, 1:1000 (Invitrogen A11122); Olig2, 1:1000 (Millipore MANB50); Islet1, 1:10 (developmental studies hybridoma bank 40.3A4-5 and 40.2D6); TUJ1 (recognizing Tubulin β III), 1:1000 (Covance MRB-435P); Microtubule Associated Protein (MAP2), 1:1000 (Chemicon MAB378); Phospho Histone H3, 1:100 (Cell Signaling 9701S). Alexa Fluor secondary antibodies were used at 1:1000. EBs cultured for 5 days were harvested by gravity, washed with PBS and paraformaldehyde-fixed, cryo-protected by sucrose, tissue tek-embedded, cut in 20 μm slices and mounted in slides for immunofluorescence protocol.
Mab378
Manufactured by Cell Signaling Technology
MAB378 is a monoclonal antibody product manufactured by Cell Signaling Technology. It is designed for use in research applications.
Automatically generated - may contain errors
Lab products found in correlation
A11122, by Merck Group (1 mentions)
Phospho histone h3, by Cell Signaling Technology (1 mentions)
Ab5603, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Mrb 435p, by Fortrea (1 mentions)
Alexa fluor 594, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
2 protocols using mab378
1
Immunofluorescence Staining of Embryoid Bodies
2
Immunostaining of Neuronal Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Neurons were washed with PBS, fixed with 4% paraformaldehyde in PBS, pH 7.3, for 10–15 min at room temperature and permeabilized in 0.05% Tween20 for 5 min at room temperature. Nonspecific binding was blocked with 5% BSA in PBS solution for 1 h. Then cells were incubated with anti-MAP2 (1:1000, Millipore, #MAB378) and anti-pCaMKII (1:1000, Cell Signaling, #33613) primary antibodies diluted in 2.5% BSA in PBS at 4 °C overnight. After three washing cycles in PBS, cells were incubated in 2.5% BSA in PBS solution with anti-rabbit (Alexa Fluor 594, 1:1000, Invitrogen, #A11012) and anti-mouse (Alexa Fluor 488, 1:1000, Invitrogen, #A11001) for 1 h at room temperature and then washed three times in PBS and visualized by a confocal microscope (ThorLabs).
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