Tissues were quick frozen in OCT (Tissue-Tek) and stored at −80°C. Tissue sections (10 µm) were fixed with ice-cold acetone for 15 min at −20°C and washed three times with PBS followed by blocking with 5% FBS. Fluorescence-labeled primary antibodies [e-flour 615 CD8 (clone 53-6.7; 1∶50), e-flour 570 ki67 (clone solA15; 1∶100; eBioscience, San Diego, CA, USA); FITC F4/80 (MCA497A488; 1∶100), Alexa Fluor 647 CD204 (MCA1322; 1∶50, AbD Serotec, Raleigh NC), Alexa Fluor 647 Gr-1 (clone RB6-8C5; 1∶100; Biolegend), FITC CD11b (clone M1/70, 1∶100; Biolegend); and 33D1 (1∶50, BD Biosciences, San Jose, CA, USA; secondary anti-rat IgG Alexa Fluor 647; Invitrogen)] were incubated with tissues overnight at 4°C in the presence of 5% FBS. Slides were then washed three times with PBS and mounted with ProLong Gold AntiFade with DAPI (Invitrogen). Images were taken using an A1 Nikon confocal microscope (Belmont, CA, USA) and percent positive cells were determined by manual counting or using Image J software (National Institute of Health) equipped with the ITCN (Image-based tool for counting nuclei) Plugin (Thomas Kuo and Jiyun Byun; Center for Bio-image Informatics at UC Santa Barbara, CA, USA) with analysis of 3–6 arbitrary regions representing randomly selected tissues from greater than one mouse per group.
Alexa fluor 647 cd204
Manufactured by Bio-Rad
Sourced in United States
Alexa Fluor 647 CD204 is a fluorescent dye conjugate used for flow cytometry analysis. It is designed to target the CD204 surface antigen, also known as the scavenger receptor class A, member 3 (SRA-3). This product provides a fluorescent label for the detection and quantification of CD204-expressing cells.
Automatically generated - may contain errors
Lab products found in correlation
E flour 615 cd8 clone 53 6.7, by Thermo Fisher Scientific (2 mentions)
E flour 570 ki67 clone sola15, by Thermo Fisher Scientific (2 mentions)
F4 80 fitc, by Bio-Rad (2 mentions)
A1 confocal microscope, by Nikon (2 mentions)
Prolong gold antifade with dapi, by Thermo Fisher Scientific (2 mentions)
Cd11b fitc clone m1 70, by BioLegend (1 mentions)
Antirabbitigg tritc, by Jackson ImmunoResearch (1 mentions)
2 protocols using alexa fluor 647 cd204
1
Quantifying Immune Cell Populations in Tissue
2
Multiparametric Immune Profiling of Frozen Tissue Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Tissues were quick frozen in OCT (Tissue-Tek)
and stored at −80 °C. Tissue sections (10 μm) were
fixed with ice-cold acetone for 15 min at −20 °C and washed
three times with 1× PBS followed by blocking with
5% fetal bovine serum in PBS. Fluorescence-labeled antibodies [e-flour
615 CD8 (clone 53-6.7; 1:50), e-flour 570 Ki-67 (clone solA15; 1:100; eBioscience, San Diego, CA, USA); FITC
F4/80 (MCA497A488;1:100); Alexa Fluor 647 CD204 (MCA1322; 1:100, AbD
Serotec, Raleigh NC, USA); 33D1 (1:100, BD Biosciences, San Jose,
CA; 1:500 secondary antirat IgG Alexa Fluor 546; Invitrogen); and
iNOS (6/iNOS/NOS; 1:100 BD Biosciences, San Jose, CA, USA; 1:500 antirabbit
IgG-TRITC, Jackson Immunoresearch Laboratories, Inc. West Grove, PA,
USA)] were incubated with tissues overnight at 4 °C in the presence
of 5% FBS. Slides were then washed three times with PBS and mounted
with ProLong Gold AntiFade with DAPI (Invitrogen). Images were taken
using an A1 Nikon confocal microscope, and the percent positive cells
were determined by manual counting of four arbitrary regions in random
samples.
and stored at −80 °C. Tissue sections (10 μm) were
fixed with ice-cold acetone for 15 min at −20 °C and washed
three times with 1× PBS followed by blocking with
5% fetal bovine serum in PBS. Fluorescence-labeled antibodies [e-flour
615 CD8 (clone 53-6.7; 1:50), e-flour 570 Ki-67 (clone solA15; 1:100; eBioscience, San Diego, CA, USA); FITC
F4/80 (MCA497A488;1:100); Alexa Fluor 647 CD204 (MCA1322; 1:100, AbD
Serotec, Raleigh NC, USA); 33D1 (1:100, BD Biosciences, San Jose,
CA; 1:500 secondary antirat IgG Alexa Fluor 546; Invitrogen); and
iNOS (6/iNOS/NOS; 1:100 BD Biosciences, San Jose, CA, USA; 1:500 antirabbit
IgG-TRITC, Jackson Immunoresearch Laboratories, Inc. West Grove, PA,
USA)] were incubated with tissues overnight at 4 °C in the presence
of 5% FBS. Slides were then washed three times with PBS and mounted
with ProLong Gold AntiFade with DAPI (Invitrogen). Images were taken
using an A1 Nikon confocal microscope, and the percent positive cells
were determined by manual counting of four arbitrary regions in random
samples.
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