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Adult c57b 6 mice

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Adult C57B/6 mice are a common inbred strain of laboratory mice. They are widely used in biomedical research due to their well-characterized genetic and physiological characteristics.

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Lab products found in correlation

Tecnai g2 spirit transmission, by Olympus (2 mentions) Vt1000s vibratome, by Leica (2 mentions) Durcupan acm epoxy resin, by Merck Group (2 mentions) Vectastain elite abc kit, by Vector Laboratories (2 mentions) Morada ccd digital camera, by Olympus (2 mentions) Cd23 biotin, by Thermo Fisher Scientific (1 mentions) Escherichia coli lps, by Merck Group (1 mentions) Anti biotin microbead, by Miltenyi Biotec (1 mentions)

3 protocols using adult c57b 6 mice

1

Isolation and Activation of Murine B Cells

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Adult C57/B6 mice were purchased from The Jackson Laboratory and maintained in a specific pathogen–free facility at the University of California, San Diego. Mice were analyzed at 8 weeks of age. Both male and female mice were used for analysis. Animal studies (S00031) were approved by the Institutional Animal Care and Use Committee. Splenocytes were incubated with CD23-biotin (eBioscience, clone B3B4) and purified using anti-biotin microbeads (Miltenyi Biotec). Sorted B cells were cultured for up to 3 days in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS), antibiotics, 2 mM l-glutamine, and 50 μM β-mercaptoethanol at 37°C and 5% CO2. LPS (Escherichia coli, Sigma-Aldrich, L2654) was used at 10 μg/ml for B cell activation.
Zhou Y, & Murre C. (2021). Bursty gene expression and mRNA decay pathways orchestrate B cell activation. Science Advances, 7(49), eabm0819.
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2

Ultrastructural Analysis of WFA-Labeled Neurons

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Adult C57B/6 mice (Jackson Labs) were transcardially perfused with 4% PFA and 0.5% glutaraldehyde (EMS) in 100 mM phosphate buffer (PB). Brains were post-fixed at 4°C overnight, and 50 μm coronal sections were cut on a Leica VT1000 S vibratome. Pre-embedding IHC was performed using WFA (Sigma L1516), amplified with Vectastain Elite ABC kit (Vector Laboratories), and developed with DAB23 (link). Sections were postfixed in 1% osmium tetroxide for 30 min and then embedded in Durcupan ACM epoxy resin (Fluka, Sigma-Aldrich)51 . For reconstruction of WFA-labeled neurons, we cut ~200 serial semithin (1.5 μm) sections on an Ultracut UC-6 ultramictrotome. Selected semithin sections were glued to resin blocks and detached from glass slides by repeated freeze-thaw. Ultrathin sections (60–80 nm) were then cut and placed on Formvar-coated single-slot grids, stained with lead citrate, and examined at 80 kV on a FEI Tecnai G2 Spirit transmission electron microscope equipped with a Morada CCD digital camera (Olympus).
Mirzadeh Z., Alonge K.M., Cabrales E., Herranz-Pérez V., Scarlett J.M., Brown J.M., Hassouna R., Matsen M.E., Nguyen H.T., Garcia-Verdugo J.M., Zeltser L.M, & Schwartz M.W. (2019). Perineuronal Net Formation during the Critical Period for Neuronal Maturation in the Hypothalamic Arcuate Nucleus. Nature metabolism, 1(2), 212-221.
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3

Ultrastructural Analysis of WFA-Labeled Neurons

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Adult C57B/6 mice (Jackson Labs) were transcardially perfused with 4% PFA and 0.5% glutaraldehyde (EMS) in 100 mM phosphate buffer (PB). Brains were post-fixed at 4°C overnight, and 50 μm coronal sections were cut on a Leica VT1000 S vibratome. Pre-embedding IHC was performed using WFA (Sigma L1516), amplified with Vectastain Elite ABC kit (Vector Laboratories), and developed with DAB23 (link). Sections were postfixed in 1% osmium tetroxide for 30 min and then embedded in Durcupan ACM epoxy resin (Fluka, Sigma-Aldrich)51 . For reconstruction of WFA-labeled neurons, we cut ~200 serial semithin (1.5 μm) sections on an Ultracut UC-6 ultramictrotome. Selected semithin sections were glued to resin blocks and detached from glass slides by repeated freeze-thaw. Ultrathin sections (60–80 nm) were then cut and placed on Formvar-coated single-slot grids, stained with lead citrate, and examined at 80 kV on a FEI Tecnai G2 Spirit transmission electron microscope equipped with a Morada CCD digital camera (Olympus).
Mirzadeh Z., Alonge K.M., Cabrales E., Herranz-Pérez V., Scarlett J.M., Brown J.M., Hassouna R., Matsen M.E., Nguyen H.T., Garcia-Verdugo J.M., Zeltser L.M, & Schwartz M.W. (2019). Perineuronal Net Formation during the Critical Period for Neuronal Maturation in the Hypothalamic Arcuate Nucleus. Nature metabolism, 1(2), 212-221.
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