PB was obtained from the retro-orbital plexus using heparinized capillary tubes (#22-362566 Fisherbrand) and lysed in red blood cell lysis buffer (R7757, Sigma-Aldrich)7 (link) employing the following antibodies: CD45.1-APC (A20) (Biolegend, San Diego, CA); B220-PECy7 and CD8-PECy7 (53-6.7) (Tonbo Biosciences, San Diego, CA); CD45.2-V500 (104), B220-PerCPCy5.5 (RA3-6B2), Gr1-PerCPCy5.5 (RB6-8C5), CD11b-PerCPCy5.5 (M1/70) and CD4-PECy7 (RM4-5) (BD Biosciences, San Diego, CA) and Ter119-PECy7 (TER-119) (Biolegend). Dilutions used and catalogue identifiers for each antibody are provided in Supplementary Table 2 .
Cd45.1 apc a20
Manufactured by BioLegend
CD45.1-APC (A20) is a fluorescently-labeled antibody that specifically binds to the CD45.1 allotypic variant of the CD45 protein. The APC (allophycocyanin) fluorescent label allows for detection and analysis using flow cytometry instrumentation.
Automatically generated - may contain errors
Lab products found in correlation
Cd8 pecy7 53 6.7, by Cytek Biosciences (2 mentions)
R7757, by Merck Group (1 mentions)
Heparinized capillary tube, by Thermo Fisher Scientific (1 mentions)
Red blood cell lysis buffer, by Merck Group (1 mentions)
Facsaria 3 sorp, by BD (1 mentions)
B220 pe cy7, by Cytek Biosciences (1 mentions)
Fitc cd45.1 a20, by BD (1 mentions)
Cd4 pecy7 rm4 5, by BD (1 mentions)
Lsrfortessa, by BD (1 mentions)
Streptavidin efluor 450, by Thermo Fisher Scientific (1 mentions)
Cd45.2 apc, by BioLegend (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Lsrfortessa flow cytometer, by BD (1 mentions)
3 protocols using cd45.1 apc a20
1
Comprehensive Blood Cell Analysis
2
Multicolor Flow Cytometry Protocol for Murine Immune Cell Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PB was collected from the retro-orbital plexus in heparinized capillary tubes (Fisherbrand, Pittsburgh, PA) and lysed in red blood cell lysis buffer (Sigma-Aldrich, St. Louis, MO). Cells were stained with the following antibodies: CD45.1-APC (A20) (Biolegend, San Diego, CA) or CD45.1-FITC (A20) (BD Biosciences, San Diego, CA), B220-PECy7 and CD8-PECy7 (53–6.7) (Tonbo Biosciences, San Diego, CA), CD45.2-V500 (104), B220- PerCPCy5.5 (RA3-6B2), Gr1-PerCPCy5.5 (RB6-8C5), Cd11b-PerCPCy5.5 (M1/70) and CD4-PECy7 (RM4-5) (BD Biosciences, San Diego, CA). All antibodies were used at 1:200 dilution. 4’,6-diamidino-2-phenylindole (DAPI) staining was used to gate live events. Analysis was performed on a LSR Fortessa and a BD FACSAria III SORP (Special Order Research Product, which contains the following LASERs: 405 nm, 445 nm, 488 nm, 562 nm, and 640 nm) (both BD Biosciences, San Diego, CA) and the data analyzed with FlowJo version 9.4.11 (Tree Star, Ashland, OR). To discern the four Confetti colors: the filter arrangements for each LASER were: CFP, 445 nm LASER—470/24 band-pass filter (BP); GFP, 488 nm LASER—515/20 BP and 505 long-pass filter (LP); YFP, 488 nm LASER—545/10 BP and 525 LP; RFP, 562 nm LASER—610/20 BP and 600 LP48 (link).
3
Flow Cytometric Analysis of Hematopoietic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H9M suspension cells were pelleted and resuspended in PBS (Gibco) supplemented with 2% FBS and 1 μM 4’,6-diamidino-2-phenylindole (DAPI; Sigma Aldrich) before flow cytometric analysis. Aliquots of BM cells from moribund mice were stained with CD11b-APC/Cy7 (M1/70), Gr1-Alexa Fluor 700 (RB6-8C5), cKit-PE/Cy7 (2B8), B220-biotin (RA3-6B2), CD3e-biotin (145-2C11), and optionally CD45.1-APC (A20) or CD45.2-APC (104) (all BioLegend), prior to staining with Streptavidin-eFluor450 (eBioscience). Data acquisition was performed in the presence of 0.5 μg/mL propidium iodide (PI). PB nucleated cells were stained with CD11b-APC/Cy7, Gr1-Alexa Fluor 700, cKit-PE/Cy7, and either CD45.1-APC or CD45.2-APC antibodies. Cells were resuspended in 1 μM DAPI for flow cytometric analysis. BM cells from short-term homing assays were flushed from the tibia, femurs, and iliac crests of recipient mice before staining with CD45.1-Pacific Blue (A20; BioLegend) and CD45.2-APC and 0.5 μg/mL PI for dead cell exclusion. 12xFGB H9M cell mixes were stained likewise and tracked over time. All data were recorded on an LSRFortessa flow cytometer (BD Biosciences), and data analyses were performed with FlowJo (Tristar).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!