Anti cd69 clone fn50

Manufactured by BioLegend

Sourced in United States

Anti-CD69 (clone FN50) is a monoclonal antibody that specifically binds to the CD69 cell surface receptor. CD69 is a type II transmembrane glycoprotein that is rapidly induced on the surface of activated T cells, B cells, and NK cells. This antibody can be used to detect and study CD69 expression.

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Lab products found in correlation

Anti cd154 clone 24 31, by BioLegend (1 mentions) Anti ki 67 clone b56, by BD (1 mentions) Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Lsr 2 flow cytometer, by BD (1 mentions) Anti myc clone 9b11, by Cell Signaling Technology (1 mentions) Anti cd45ra clone hi100, by BioLegend (1 mentions) Percp cy5, by BD (1 mentions) Anti cd25 clone bc96, by BioLegend (1 mentions) Fluorescein isothiocyanate (fitc), by R&D Systems (1 mentions)

3 protocols using anti cd69 clone fn50

Activation-Induced Marker (AIM) Staining of Antigen-Specific T Cells

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After peptide stimulation, the cells were washed with PBS containing 5% FCS and stained with amine-reactive viability stain (Live/dead fixable aqua dead cell stain kit; Invitrogen) for 30 min at 4°C. After washing, pMHC-spheromers were added to screen the epitope-specific CD8⁺ and CD4⁺ T cells. Meanwhile, the antibody cocktail was added for AIM staining (anti-CD69 (clone FN50, Biolegend), anti-CD154 (clone 24-31, Biolegend), anti-CD137 (clone 4B4-1, Biolegend), anti-CD38 (clone HIT2, BD Biosciences) and anti-Ki-67 (clone B56, BD Biosciences)). The cells were stained for 30 min at 4°C in 100μl volume.
Cells were then washed twice with staining buffer before acquisition using BD LSRII flow cytometer. The data was analyzed using FlowJo (v10) software.

Gao F., Mallajoysula V., Arunachalam P.S., van der Ploeg K., Manohar M., Röltgen K., Yang F., Wirz O., Hoh R., Haraguchi E., Lee J.Y., Willis R., Ramachandiran V., Li J., Kathuria K.R., Li C., Lee A.S., Shah M.M., Sindher S.B., Gonzalez J., Altman J.D., Wang T.T., Boyd S.D., Pulendran B., Jagannathan P., Nadeau K.C, & Davis M.M. (2023). Spheromers reveal robust T cell responses to the Pfizer/BioNTech vaccine and attenuated peripheral CD8+ T cell responses post SARS-CoV-2 infection. Immunity, 56(4), 864-878.e4.

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Multiparameter Flow Cytometry Immunophenotyping

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Cells were washed with 1X PBS (Sigma) supplemented with 0.5% bovine serum albumin (RPI) and 2 mM EDTA, then surface stained by incubating with antibodies for 15 minutes on ice. They were subsequently washed again prior to flow analysis on a BD Accuri C6 or Beckman Cytoflex S or cell sorting with a BD FACSAria II or Sony MA900. Anti-CD4 (clone SK3), anti-CD8 (clone SK1), anti-PD-1 (clone EH12.2H7), anti-TIM3 (clone F38-2E2), anti-LAG3 (clone 11C3C65), anti-CD3 (clone OKT3), anti-CD62L (clone DREG-56), anti-CD45RA (clone HI100), and anti-CD69 (clone FN50) antibodies were purchased from Biolegend. Anti-Myc (clone 9B11) and anti-Flag (D6W5B) antibodies were from Cell Signalling Technology.

Gordon K.S., Kyung T., Perez C.R., Holec P.V., Ramos A., Zhang A.Q., Agarwal Y., Liu Y., Koch C., Starchenko A., Joughin B.A., Lauffenburger D.A., Irvine D.J., Hemann M.T, & Birnbaum M.E. (2022). Screening for CD19-specific chimaeric antigen receptors with enhanced signalling via a barcoded library of intracellular domains. Nature biomedical engineering, 6(7), 855-866.

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Multiparametric Flow Cytometry of Lymphocyte Populations

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The following labelled murine anti-human monoclonal antibodies were used
to delineate lymphocyte populations by flow cytometry analysis: anti-CD3 (clone
UCHT1l) labeled with phycoerythrin-Texas Red-X (ECD) (Beckman Coulter, USA),
anti-CD4 (clone RPA-T4) labeled with V500 (BD Horizon, USA), anti-CD25 (clone
BC96) labeled with allophycocyanin with cyanin-7 (APCcy7) (Biolegend, USA),
anti-CD127 (clone A19D5) labeled with brilliant violet 421 (BV421) (Biolegend),
anti- FoxP3 (clone 236a) labeled with peridinin chlorophyll protein complex with
cyanin-5.5 (PerCP-Cy5.5) (BD Pharmigen), anti-CD39 (clone eBioA1) labeled with
phycoerythrin (PE) (eBioscience, USA), anti-CTLA-4 (clone L3D10) labeled with
allophycocyanin (APC) (Biolegend), anti-CD69 (clone FN50) labeled with
phycoerythrin-carbocyanin (PE-Cy7) (Biolegend), anti-HLA-DR (clone L243) labeled
with Alexa 700 (BD Pharmigen), anti-TNFRII (clone 22235) labeled with
fluorescein isothiocyanate (FITC) (R&D Systems, USA), anti-OX40 (clone
ACT35) labeled with PE-Cy7 (Biolegend), anti-GITR (110416) labeled with FITC
(R&D Systems), anti-CD45RO (clone UCHL1) labeled with APC (Biolegend),
and anti-CD45RA labeled with Alexa 700 (Biolegend). Antibody panels used in this
study are presented in Supplementary Table 1. We titrated each antibody and selected its
final dilution for an optimal specific staining associated with a low
background.

Gonçalves-Lopes R.M., Lima N.F., Carvalho K.I., Scopel K.K., Kallás E.G, & Ferreira M.U. (2016). Surface expression of inhibitory (CTLA-4) and stimulatory (OX40) receptors by CD4+ regulatory T cell subsets circulating in human malaria. Microbes and infection, 18(10), 639-648.

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