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Anti cd45ro pecy7

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Anti-CD45RO PECy7 is a monoclonal antibody that binds to the CD45RO antigen on the surface of human T cells. It is conjugated with the PECy7 fluorescent dye.

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Lab products found in correlation

Celltrace violet, by Thermo Fisher Scientific (2 mentions) Anti cd19 pe, by BioLegend (1 mentions) Anti cd3 bv711, by BioLegend (1 mentions) Anti cd27 pe, by BioLegend (1 mentions) Anti cd45ra bv421, by BioLegend (1 mentions) Anti cd8 bv711, by BioLegend (1 mentions) Streptavidin pe, by BioLegend (1 mentions) Anti ccr7 apc, by BioLegend (1 mentions) Anti cd45 bv711, by BioLegend (1 mentions) Anti cd4 bv785, by BioLegend (1 mentions) Fortessa flow cytometer, by BD (1 mentions) Alexa fluor 647, by Jackson ImmunoResearch (1 mentions) Anti cd8 fitc, by BioLegend (1 mentions) Anti cd137 pecy7, by BioLegend (1 mentions) Anti cd45ra percp cy5, by BioLegend (1 mentions) Anti cd3 percp cy5, by BioLegend (1 mentions) Anti ccr7 apc cy7, by BioLegend (1 mentions) Anti bovine igg, by Merck Group (1 mentions) True nuclear transcription factor buffer set, by BioLegend (1 mentions) Anti cd8 v500, by BD (1 mentions)

4 protocols using anti cd45ro pecy7

1

Comprehensive CAR Expression Analysis

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For evaluation of CAR expression, cells were stained with a goat anti-mouse Fab antibody conjugated with Alexa Fluor 647 (Jackson ImmunoResearch) or biotinylated CD19-Fc (ACRO Biosystems) for 15 min at room temperature. Cells were thoroughly washed before staining with antibodies for additional surface markers. Anti-CD3-BB515 and anti-CD62L-BB515 were purchased from BD Biosciences. Anti-CD4-BV785, anti-CD8-BV711, anti-CD45RA-BV421, anti-CD45RO-PECy7, streptavidin-PE, anti-CD27-PE, anti-CD95-allophycocyanin (APC)Cy7, anti-CCR7-APC, anti-CD45-BV711, and anti-CD19-PE, anti-CD3-BV711 were purchased from BioLegend and used to stain surface antigens. Cell Trace Violet was purchased from Thermo Fisher Scientific and used at a concentration of 1 μM to label cells for 10 min. All data were acquired using a BD Fortessa flow cytometer (BD Biosciences) and analyzed using FlowJo software.
MacLeod D.T., Antony J., Martin A.J., Moser R.J., Hekele A., Wetzel K.J., Brown A.E., Triggiano M.A., Hux J.A., Pham C.D., Bartsevich V.V., Turner C.A., Lape J., Kirkland S., Beard C.W., Smith J., Hirsch M.L., Nicholson M.G., Jantz D, & McCreedy B. (2017). Integration of a CD19 CAR into the TCR Alpha Chain Locus Streamlines Production of Allogeneic Gene-Edited CAR T Cells. Molecular Therapy, 25(4), 949-961.
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2

Multiparametric Immune Cell Phenotyping

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Anti-HLA-A*02 APC (RRID AB 2561567, clone BB7.2), anti-CD3 PerCPCy5.5 (RRID AB 2561628, clone OKT3) and PE (RRID AB 571913, clone OKT3), anti-CD8 FITC (RRID AB 1877178, clone SK1), anti-CD137 PECy7 (RRID AB 2207741, clone 4B4-1), anti-CD45RO PECy7 (RRID AB 11203903, clone UCHL1), anti-CD45RA PerCPCy5.5 (RRID AB 893358, clone HI100), anti-CCR7 APC-Cy7 (RRID AB 10915272, clone G043H7) were bought from Biolegend (San Diego, CA, USA).
Quiros-Fernandez I., Poorebrahim M., Fakhr E, & Cid-Arregui A. (2021). Immunogenic T cell epitopes of SARS-CoV-2 are recognized by circulating memory and naïve CD8 T cells of unexposed individuals. EBioMedicine, 72, 103610.
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3

CAR T Cell Phenotyping and Transcription Factor Analysis

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GPC3-CAR expression was detected using the anti-F(ab)2 Alexa Fluor® 647-conjugated antibody (Jackson ImmunoResearch) and anti-goat IgG1 isotype control (Jackson ImmunoResearch, West Grove, PA). The following antibodies were used for T cell phenotyping analyses: anti-CD4-APC/Fire 750 (BioLegend), anti-CD8-V500 (BD Biosciences, San Jose, CA), anti-CD45RO-PE/Cy7 (BioLegend), anti-CD62L-AF488 (BioLegend), anti-CD19-PerCP/Cy5.5 (CCR1; BioLegend), anti-CD3-PE (BD Biosciences), anti-CD279-PerCP/Cy5.5 (PD-1; BioLegend), anti-CD223-PE/Cy7 (LAG-3; BioLegend) and anti-CD366-BV421 (TIM-3; BD Biosciences). Anti-bovine IgG (Sigma-Aldrich, St. Louis, MO) was used to block non-specific binding of other murine antibodies following CAR staining. Flow cytometry assessment was performed on either an LSR-II (BD Biosciences) or iQue Screener PLUS (Intellicyt Corporation, Albuquerque, NM). Results were analyzed using FlowJo software (FlowJo, Ashland, OR). To detect intracellular TCF-1 expression, CAR T cells were first stained for surface expression of CAR, CD4, and CD8 using the antibodies listed above, followed by staining with anti-TCF1-PE (TCF7, BioLegend) used in conjunction with the True-Nuclear™ Transcription Factor Buffer Set (BioLegend) according to the manufacturer’s instructions.
Batra S.A., Rathi P., Guo L., Courtney A.N., Fleurence J., Balzeau J., Shaik R.S., Nguyen T.P., Wu M.F., Bulsara S., Mamonkin M., Metelitsa L.S, & Heczey A. (2020). Glypican-3-specific CAR T cells co-expressing IL15 and IL21 have superior expansion and antitumor activity against hepatocellular carcinoma. Cancer immunology research, 8(3), 309-320.
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4

Detailed Phenotyping of Immune Cells

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Anti-CD14 and anti-CD19 APC-H7 (both at 1:100), anti-CD25 PECy7 (1:50), anti-CD62L V450 (1:50), anti-CD127 PECy7 (1:100), anti-CD161 FITC (1:10), anti-IFNγ V450 (1:200), anti-Ki67 FITC (1:10) were from BD Biosciences. Anti-CD3 ECD (1:100) and anti-CD8β PE (1:50) were from Beckman Coulter. Anti-Vα7.2 FITC (1:10) and PE (1:20), anti-CD45 Alexa Fluor 700 (1:400), anti-CD45RO PECy7 (1:50), anti-CCR9 Alexa Fluor 647 (1:20), anti-Ki67 Brilliant Violet 421 (1:20), anti-CCR7 Brilliant Violet 421 (1:20) were from BioLegend. Anti-CD161 PerCP-Cy5.5 (1:20), anti-IL-22 PerCP-eFluor 710 (1:50) were from eBioscience. Anti-IL-18R PE (1:20), and anti-PLZF APC (1:10) were from R&D systems. Anti-CD4 Qdot 655 (1:400), anti-CD8α Qdot 605 (1:800), anti-CD45 Qdot 705 (1:100), live/dead aqua (1:100) and near infrared fixable (1:400) cell stain were from Invitrogen. Cell surface staining was performed using directly conjugated antibodies and fixed in Cytofix/Cytoperm (BD Biosciences). Intracellular staining was performed using the appropriate mAb’s in Perm/Wash (BD Biosciences). Samples were acquired on an LSRII flow cytometer (BD Biosciences) equipped with 405, 488, 532 and 647 nm lasers. Single-stained polystyrene beads (BD Biosciences) were used for compensation purposes. Software-based compensation was performed using the compensation platform in FlowJo software version 9.5 (Tree Star).
Leeansyah E., Loh L., Nixon D.F, & Sandberg J.K. (2014). Acquisition of innate-like microbial reactivity in mucosal tissues during human fetal MAIT-cell development. Nature Communications, 5, 3143.
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