The GM from a biopsy punch (Ø 0.25 cm) around the lesion area was dissociated using the adult brain dissociation kit mouse and rat from (MiltenyiBiotec) following the manufacturer's instructions except the red blood cell removal step. This was not necessary as the meninges were removed during tissue preparation. Immune cells were isolated by CD45 magnetic associated cell separation (MACS) according to the manufacturer's protocol (anti‐CD45 MicroBead Kit, mouse; Miltenyi) and then stained with the following antibodies for flow cytometry (Biolegend): anti‐Gr‐1 (BV421; 1:500); anti‐ Ly6c (PE) anti‐CD45 APCCy7 (clone 30‐F11; 1:100); anti‐CD11b BV421 (clone M1/70; 1:500); anti‐CD3 Alexa‐488 (clone 145‐2C11; 1:400); anti‐CD19 Alexa‐647 (clone 6D5; 1:500); and anti F4/80 PeCy7 (clone BM8; 1:500). FACS measurements were performed on a FACS Canto II (BD), and data were analyzed using FlowJo software.
Anti cd11b bv421 clone m1 70
Manufactured by BioLegend
Anti-CD11b BV421 (clone M1/70) is a fluorescently-labeled antibody that binds to the CD11b cell surface antigen. CD11b is a subunit of the integrin Mac-1 (CD11b/CD18) and is expressed on the surface of myeloid cells, including monocytes, macrophages, and granulocytes. This antibody can be used in flow cytometry applications to identify and characterize myeloid cell populations.
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Lab products found in correlation
Facscanto 2, by BD (2 mentions)
Anti f4 80 pe cy7 clone bm8, by BioLegend (1 mentions)
Anti cd25 pe clone pc61, by BioLegend (1 mentions)
Anti f4 80 pe bm8, by BioLegend (1 mentions)
Anti cd11b fitc clone m1 70, by BioLegend (1 mentions)
Anti cd27 apc clone l g 3a10, by BioLegend (1 mentions)
Anti cd8 fitc clone 53.6.7, by BioLegend (1 mentions)
Anti cd3 bv421 clone 17a2, by BioLegend (1 mentions)
Fortessa, by BD (1 mentions)
Apc cy7 anti cd45 30 f11, by BioLegend (1 mentions)
2 protocols using anti cd11b bv421 clone m1 70
1
Immune cell profiling from brain biopsy
2
Multiparameter Flow Cytometry Analysis
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For FACS analysis, the following antibodies were used: anti-CD3-Bv421-(clone 17A2, Biolegend), anti-CD4-Bv421 (clone RM 4-5, eBioscience), anti-CD8-FITC (clone 53-6.7, Biolegend), anti-CD25 PE (clone PC61, Biolegend), anti-CD27-APC (clone LG.3A10, Biolegend), anti-CD11b-FITC (clone M1/70, Biolegend), anti-CD11b-Bv421 (clone M1/70, Biolegend), anti-FceRIα-PE (MAR-1, Biolegend), anti-F4/80-PE (BM8, Biolegend), anti-CD45-APC/Cy7 (30-F11, Biolegend), anti-CD107a-FITC (1D4B, Biolegend) and anti-P2X7-AF647 (clone Hano44, UKE) (Adriouch et al., 2005) (link). Flow cytometric analyses were performed on a BD Fortessa (Beckton Dickinson) or a BD FACS CantoII (Beckton Dickinson).
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