Live fluorescence immunostaining was performed as previously reported.44 (link) Briefly, live slices were transferred placed on parafilm, Fc-blocked for 30 min, and labelled with an antibody cocktail for one hour. Antibodies used were FITC-rat anti-mouse/human CD45R/B220 (clone RA3–6B2, BioLegend), and rat anti-mouse CD4 F(ab’)2 fragment (clone GK1.5, BioLegend, fragmented according to published procedures45 (link)) that was conjugated using AlexaFluor 594-succinimidyl ester according to manufacturer instructions (Thermo Fisher). All antibodies were used at a concentration of 20 μg/mL (0.2 μg per tissue slice). Slices were rinsed for 1 hr in PBS to remove unbound antibodies.
Fitc rat anti mouse human cd45r b220 clone ra3 6b2
Manufactured by BioLegend
FITC-rat anti-mouse/human CD45R/B220 (clone RA3–6B2) is a fluorescently labeled monoclonal antibody. It binds to the CD45R/B220 antigen expressed on B cells and a subset of T cells.
Automatically generated - may contain errors
2 protocols using fitc rat anti mouse human cd45r b220 clone ra3 6b2
1
Live Fluorescence Immunostaining of Tissue Slices
2
Live Immunostaining of Brain Slices
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Live fluorescence immunostaining was performed as previously reported. 44 (link) Briefly, live slices were transferred placed on parafilm, Fc-blocked for 30 min, and labelled with an antibody cocktail for one hour. Antibodies used were FITC-rat anti-mouse/human CD45R/B220 (clone RA3-6B2, BioLegend), and rat anti-mouse CD4 F(ab')2 fragment (clone GK1.5, BioLegend, fragmented according to published procedures 45 (link) ) that was conjugated using AlexaFluor 594-succinimidyl ester according to manufacturer instructions (Thermo Fisher). All antibodies were used at a concentration of 20 μg/mL (0.2 μg per tissue slice). Slices were rinsed for 1 hr in PBS to remove unbound antibodies.
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