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Mononuclear cells from the spinal cord were isolated following a published protocol by Korn T et al [54].
The spleen and the spinal cord were isolated at indicated time points after CFA immunization.
Cells from lymph nodes and spinal cord were isolated as described before [ 26].
Brain and spinal cord were isolated and fixed in 4% paraformaldehyde and dehydrated in a 30% sucrose solution.
Mononuclear cells from the brain or spinal cord were isolated by enzymatic digestion of tissues with collagenase D (0.2 mg/ml) and DNase I (0.2 mg/ml) followed by Percoll® gradient centrifugation.
The L4 L6 segments of the lumbar spinal cord were isolated and homogenized in a lysis buffer containing protease inhibitor (Sigma); subsequently 20 mg of proteins were loaded for each lane and separated by SDS-PAGE gel (7.5%), and transferred to PVDF membrane (Bio-Rad).
Similar(53)
To accomplish this we used an in vitro preparation of the mouse where the spinal cord was isolated (T5-cauda equina) with one hind limb left attached.
Embryos were allowed to develop until E8 at which point the spinal cord was isolated to assess GFP expression.
The spinal cord was isolated by laminectomy and post-fixed in 4% PFA for 2 hours at 4°C.
The lumbar spinal cord was isolated at E8, fixed in Zamboni's fixative overnight at 4°C, cryoprotected in 30% sucrose and embedded in OCT medium (Tissue-Tek) before freezing and cryostat sectioning into 14 µm-slices.
The lumbar section of the spinal cord was isolated and placed in 4% PFA overnight.
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