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Klrg1 apc

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KLRG1-APC is a fluorochrome-conjugated antibody that binds specifically to the KLRG1 (Killer Cell Lectin-Like Receptor G1) surface marker. KLRG1 is expressed on a subset of natural killer (NK) cells, T cells, and other cytotoxic lymphocytes. The APC (Allophycocyanin) fluorescent dye is conjugated to the antibody, allowing for the detection and quantification of KLRG1-positive cells using flow cytometry.

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Lab products found in correlation

Cd44 percp cy5, by Thermo Fisher Scientific (1 mentions) Cd8 alexa fluor 700, by Thermo Fisher Scientific (1 mentions) Cd127 fitc, by Thermo Fisher Scientific (1 mentions) Anti cd4 fitc, by Thermo Fisher Scientific (1 mentions) Flow count beads, by Beckman Coulter (1 mentions) Anti pd 1 fitc, by Thermo Fisher Scientific (1 mentions) Anti cd3 efluor 450, by Thermo Fisher Scientific (1 mentions) Anti cd8a percp cy5, by Thermo Fisher Scientific (1 mentions) Facscanto 2, by BD (1 mentions) Facsdiva, by BD (1 mentions) Golgiplug, by BD (1 mentions) Foxp3 staining buffer kit, by Thermo Fisher Scientific (1 mentions) Foxp3 pe, by Thermo Fisher Scientific (1 mentions) Cd11b percp cy5, by Thermo Fisher Scientific (1 mentions) Cd25 pe cy7, by Thermo Fisher Scientific (1 mentions) Cd4 percp cy5, by Thermo Fisher Scientific (1 mentions) Cd3 fitc, by Thermo Fisher Scientific (1 mentions) Cd62l efluor450, by Thermo Fisher Scientific (1 mentions) Cytoflex, by BD (1 mentions) F4 80 pe cy7, by Thermo Fisher Scientific (1 mentions)

4 protocols using klrg1 apc

1

Phenotyping OVA-specific CD8+ T cells

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Splenocytes prepared as described previously were stained with H-2Kb-OVA257 MHC I pentamers (ProImmune, Oxford, UK) in room temperature FACS buffer for 10 minutes in the presence of 2-4G2 antibody. Cells were washed once and stained with surface antibodies plus L/D NIR for 20 minutes on ice. Antibodies included CD127-FITC, CD44-PerCP-Cy5.5, KLRG1-APC, CD8-Alexa Fluor 700 (all from eBioscience), and B220-V500 (BD). Cells were washed, fixed with Cytofix, and analyzed as previously. Within the viable CD8 T-cell population, CD44hi H-2Kb-OVA257 pentamer+ events, gated based on the 99.9th percentile in unimmunized mice, were analyzed for their expression of KLRG1 and CD127.
Odegard J.M., Kelley-Clarke B., Tareen S.U., Campbell D.J., Flynn P.A., Nicolai C.J., Slough M.M., Vin C.D., McGowan P.J., Nelson L.T., ter Meulen J., Dubensky TW J.r, & Robbins S.H. (2015). Virological and Preclinical Characterization of a Dendritic Cell Targeting, Integration-deficient Lentiviral Vector for Cancer Immunotherapy. Journal of Immunotherapy (Hagerstown, Md. : 1997), 38(2), 41-53.
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2

Multiparametric flow cytometry analysis

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Cell enumeration was performed in whole blood samples using Flow-Count™ beads (Beckman Coulter, UK) according to manufacturer’s instructions. After red blood cell lysis, mononuclear cells were stained with anti-CD3/eFluor 450, anti-CD4/FITC and anti-CD8a/PerCP-Cy5.5 monoclonal antibodies (mAb) (all eBioscience, USA). Expression of PD-1, KLRG1 and LAG-3 was assessed in whole blood samples after staining with anti-CD3/eFluor 450, anti-CD8a/PerCP-Cy5.5, anti-PD-1/FITC, KLRG-1/APC and anti-LAG-3/PE mAbs (all eBioscience). Pentamer analysis was performed as previously described [14 (link)], using H-2Kb/SIINFEKL or H-2Kb/SVYDFFVWL Pro5 pentamer/PE (ProImmune, UK).
To assess peptide-induced intracellular accumulation of IFN-γ by CD8+ T cells, splenocytes were stimulated with 1 μg/mL SVL peptide, 0.5 μg/mL co-stimulatory anti-CD28 antibody (eBioscience) in the presence of GolgiPlug (BD Biosciences, Belgium) for 5 h prior to fixation, permeabilization, and staining with anti-CD3/eFluor 450, anti-CD8a/PerCP-Cy5.5 and anti-IFN-γ/PE mAbs (eBioscience).
Samples were analysed using a FACSCantoII (BD Biosciences) and FACSDiva (BD Biosciences) or FlowJo (Treestar, OR) software.
Tye G.J., Ioannou K., Amofah E., Quartey-Papafio R., Westrop S.J., Krishnamurthy P., Noble A., Harrison P.M., Gaensler K.M., Barber L.D, & Farzaneh F. (2015). The combined molecular adjuvant CASAC enhances the CD8+ T cell response to a tumor-associated self-antigen in aged, immunosenescent mice. Immunity & Ageing : I & A, 12, 6.
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3

Multiparametric Flow Cytometry Analysis

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Cells were incubated with fixable viability dye eFluor780 for 10 minutes at 4°C and washed twice in FACS buffer prior to staining in order distinguish live cells from the dead cells. Cells were incubated with Fc-blocking antibodies for 10 minutes at 4°C, then incubated for 30 minutes at 4°C with CD3 FITC, CD4 PerCP Cy5.5, CD25 PE-Cy7, CD62LeFluor450, FOXP3 PE, KLRG1APC, CD127 AF700, CD11B PerCp Cy5.5, CD11cAPC, GR1 PE, F4/80 PE-Cy7 from eBioscience, and CD8 BV605 and CD44 BV510 from Biolegend. For intracellular staining, cells were fixed and permeabilized with FOXp3 Staining Buffer Kit (eBioscience) according to manufacturer’s instructions. Cells were incubated for 30 minutes at room temperature in dark with Foxp3-PE and analyzed using the CytoFLex (BD Biosciences) and Kaluza software (Beckman Coulter).
Schrand B., Verma B., Levay A., Patel S., Castro I., Benaduce A.P., Brenneman R., Umland O., Yagita H., Gilboa E, & Ishkanian A. (2017). Radiation-Induced Enhancement of Antitumor T-cell Immunity by VEGF-Targeted 4–1BB Costimulation. Cancer research, 77(6), 1310-1321.
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4

Multiparameter Flow Cytometry Analysis

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Cells were stained with fluorochrome-conjugated monoclonal antibodies and 2.4G2 blocking antibody and incubated in the dark for 30 min on ice. For CD1d tetramer staining, cells were incubated in the dark for 15 min at room temperature followed by 15 min on ice. Flow cytometric analysis was performed on a FACSAria (BD Biosciences) or a MACSQuant (Miltenyi Biotec). Data were analyzed using FlowJo (Tree Star).
TCRβ-FITC, HSA-FITC, CD4-FITC, TCRβ-PE, Ly49C/I/F/H-PE, TCRβ-PErCPCy5.5, Ly49G2-PerCPeFluor710, CD62L-PE-Cy6, NK1.1-PE-Cy7, KLRG1-APC, CD25-APC, CD19-PE, CD44-APCeFluor780, CD8α-eFluor450, and CD3-eFluor450 were purchased from ebioscience. Ly49G2-FITC, Ly49C/I-FITC, Ly49A/D-PE, and CD4-PerCP were purchased from BD Pharmingen. BV421-CD127, BV510-TCRβ, BV570-CD45, BV605-CD3, BV605-CCR6, and BV785-NK1.1 were purchased from Biolegend. PE- and APC-conjugated α-GalCer loaded CD1d tetramer was prepared in our laboratory. CD1d, M45 and M57 tetramers were kindly provided by the National Institute of Allergy and Infectious Disease MHC Tetramer Core Facility at Emory University (Atlanta, GA, USA).
Miah S.M., Jayasuriya C.T., Salter A.I., Reilly E.C., Fugere C., Yang W., Chen Q, & Brossay L. (2017). Ptpn11 Deletion in CD4+ Cells Does Not Affect T Cell Development and Functions but Causes Cartilage Tumors in a T Cell-Independent Manner. Frontiers in Immunology, 8, 1326.
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