PBS (Invitrogen, Carlsbad, CA, USA) and separated by Ficoll-Paque (GE Healthcare, Piscataway, NJ, USA) centrifugation. Mouse splenocytes were isolated by disaggregation and 70μm filtration (BD, San Diego, CA, USA) to form a single cell suspension. Cells were enumerated using a Z™ Series Coulter Counter (Beckman Coulter Inc., NSW, Australia).
70μm filtration
Manufactured by BD
Sourced in United States
The 70μm filtration is a lab equipment product that is designed to filter particles down to a size of 70 micrometers. It is used to separate and isolate components of a sample based on their size.
Automatically generated - may contain errors
Lab products found in correlation
Z series coulter counter, by Beckman Coulter (4 mentions)
Ficoll paque, by GE Healthcare (4 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (4 mentions)
Rbc lysis buffer, by Thermo Fisher Scientific (2 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Percoll, by GE Healthcare (1 mentions)
Dnase 1, by Merck Group (1 mentions)
5 protocols using 70μm filtration
1
Mouse Splenocyte Isolation and Enumeration
2
Mouse Splenocyte Isolation and Enumeration
3
Microglia Isolation and Characterization
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At 6 h after LPS treatment, microglia cells from the adenovirus injected groups were isolated from whole brain homogenates by Percoll gradient centrifugation according to previous reports46 (link),57 (link),58 . Briefly, brain tissue was gently dissociated by grinding in a 15-ml dounce homogenizer (Glass Potter, Braun, Melsungen, Germany) containing DNase I (0.025 U ml–1 final concentration, Sigma) and passed through 70-μm filtration (BD Biosciences, Boston, MA). After centrifugation, the supernatants were discarded, and the cell pellets were resuspended in 40% Percoll (GE-healthcare, Uppsala, Sweden), followed by spining at 800 g for 30 min with discontinuous Percoll gradients (HBSS/30%/40%/70%). Microglia cells were collected at the 40%/70% interface. Cells were washed and then resuspended in FACS buffer (1X PBS with 2% FBS). Quantification of CD11b + cells with CD45, MHC class II, and CD86 was performed with a FACSCalibur flow cytometer (Becton Dickinson) using standard procedures.
4
Purification and Enumeration of pDCs
5
Purification and Enumeration of pDCs
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Lymphocytes from HD and CLL patients were purified from peripheral blood diluted 1:1 in PBS (Invitrogen, Carlsbad, CA, USA) and layered onto Ficoll-Paque (GE Healthcare, Piscataway, NJ, USA) for gradient centrifugation. Mouse splenocytes were isolated following disaggregation of the spleen and 70μm filtration (BD, San Diego, CA, USA) to form a single cell suspension. Red blood cells were lysed with RBC lysis buffer (eBioscience, San Diego, CA, USA). Cells were enumerated using a Z™ Series Coulter Counter (Beckman Coulter Inc., NSW, Australia). Absolute numbers of pDCs were calculated by multiplying the percentage of FACS-gated pDCs by total cell count.
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