Flow cytometry analysis was performed at the NPC and the final neuronal stages. Cells were harvested by enzymatic dissociation using Accutase for 5 min at 37°C until cells were visibly detached. Cells were pelleted by centrifugation at 800 rpm for 3 minutes, resuspended in ice-cold PBS. Cells were fixed in ice-cold 1%PFA in PBS and permeabilized in 0.1% TritonX-100 in PBS. Cells were blocked in 10% Rabbit serum (Abcam) and 10% Mouse serum (Abcam) for 1h. Cells were then resuspended in blocking buffer containing the appropriate concentration of conjugated antibody according to the manufacturer’s specifications (see Supplementary Table S3 for the list of the antibodies used in this study). Cells were sorted with Becton Dickinson Canto II, and Becton Dickinson FacsCalibur, followed by analysis using FlowJo 10.2 (FlowJo, LLC)
Mouse serum
Manufactured by Abcam
Sourced in United Kingdom
Mouse serum is a biological fluid obtained from the blood of mice. It contains a complex mixture of proteins, enzymes, hormones, and other biomolecules that are present in the circulatory system of the animal.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using mouse serum
1
Neuronal Differentiation Flow Cytometry
2
Chsy1 Mutant Bone Marrow and Spleen Analysis
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Bone marrow from femurs and spleen were collected from 8-week-old Chsy1skt mutants and heterozygous controls for flow cytometry. Cells were washed with PBS, blocked with 5% mouse serum (abcam, Cambridge, UK) for 15 min, stained with fluorochrome-conjugated antibodies (Table S3) for 30 min at 4°, washed, resuspended in PBS, and analyzed by flow cytometry (BD LSR II cytometer; BD Biosciences, San Jose, CA) with FlowJo analysis software (version 9.3.1; FlowJo, Ashland, OR).
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