Cd86 2331 fun 1

Manufactured by BD

Sourced in United States

CD86 (2331 (FUN-1)) is a laboratory product. It is a protein that plays a role in the immune system. The core function of this product is to facilitate the interaction between antigen-presenting cells and T cells.

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Lab products found in correlation

Cd83 hb15e, by BD (2 mentions) Bsa pbs, by Merck Group (2 mentions) Cd80 l307.4, by BD (2 mentions) Facscanto flow cytometer, by BD (2 mentions) Granzyme b, by BD (1 mentions) Fixable viability dye efluor 780, by Thermo Fisher Scientific (1 mentions) Perm wash solution, by BD (1 mentions) Donkey anti goat a21447, by Thermo Fisher Scientific (1 mentions) Ace2 af933, by R&D Systems (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Cd11b m1 70, by BD (1 mentions) Cd45 30 f11, by BD (1 mentions) Cd31 mec 13, by BD (1 mentions) Cd44 im7, by Thermo Fisher Scientific (1 mentions) C kit 2b8, by BioLegend (1 mentions) Cd34 mec14, by BioLegend (1 mentions) Sca1 d7, by BioLegend (1 mentions) Cd80 16 10a1, by BD (1 mentions) Hmβ1 1, by BioLegend (1 mentions)

4 protocols using cd86 2331 fun 1

Measuring Immune Cell Activation and Cytotoxicity

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To measure DC activation markers, cells were fixated for 30 min with 4% paraformaldehyde (PFA, Electron Microscopy Sciences). For cell surface staining, cells were incubated in 0.5% PBS-BSA (Sigma-Aldrich) containing antibodies for 30 min at 4°C. To measure CTL responses, PBLs were harvested and stained for viability using Fixable Viability Dye eFluor 780 (eBioscience) for 5 min at 4°C and subsequently fixated for 10 min with 2% PFA. After fixation, cells were permeabilized using Perm/Wash solution (BD Biosciences) for 5 min 4°C and incubated with antibodies diluted in Perm/Wash solution for 10 min at 4°C. Single-cell measurements were performed with a FACS Canto flow cytometer (BD Biosciences). FlowJo V.10 software was used to analyze the data. Antibody clones used to analyze DC activation are CD86 (2331 (FUN-1)), CD80 (L307.4), and CD83 (HB15e) (all BD Pharmingen). For each experiment, live cells were gated on FSC (forward scatter) and SCC (side scatter) and analyzed further with the markers mentioned. Antibody clones used for cytotoxic T-cell responses are CD3 (UCHT1), CD8 (RPA-T8), IFN-γ (B27) (all BioLegend), perforin (dG9, eBioscience), and granzyme B (GB11, BD Pharmingen). For each experiment, live cells were selected and gated on CD3 and CD8 expressions. In this CD3⁺CD8⁺ population, cytokine expression was analyzed.

Waanders L., van der Donk L.E., Ates L.S., Maaskant J., van Hamme J.L., Eldering E., van Bruggen J.A., Rietveld J.M., Bitter W., Geijtenbeek T.B, & Kuijl C.P. (2023). Ectopic expression of cGAS in Salmonella typhimurium enhances STING-mediated IFN-β response in human macrophages and dendritic cells. Journal for Immunotherapy of Cancer, 11(4), e005839.

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Cell Surface Marker Analysis by Flow Cytometry

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For cell surface staining, cells were incubated in 0.5% PBS‐BSA (Sigma‐Aldrich) containing antibodies for 30 min at 4°C. Single‐cell measurements were performed on a FACS Canto flow cytometer (BD Biosciences) and FlowJo V10 software (TreeStar) was used to analyze the data. The antibody clones used are: CD86 (2331 (FUN‐1), BD Pharmingen), CD80 (L307.4, BD Pharmingen), CD83 (HB15e, BD Pharmingen), ACE2 (AF933, R&D Systems), goat‐IgG (AB‐2535864, ThermoFisher Scientific), donkey‐anti‐goat (A‐21447, ThermoFisher Scientific). For each experiment, live cells were gated on FSC and SSC and analyzed further with the markers mentioned (Supporting information Fig. S1). The authors adhered to the guidelines for the use of flow cytometry and cell sorting in immunological studies [37 (link)].

van der Donk L.E., Eder J., van Hamme J.L., Brouwer P.J., Brinkkemper M., van Nuenen A.C., van Gils M.J., Sanders R.W., Kootstra N.A., Bermejo‐Jambrina M, & Geijtenbeek T.B. (2022). SARS‐CoV‐2 infection activates dendritic cells via cytosolic receptors rather than extracellular TLRs. European Journal of Immunology, 52(4), 646-655.

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Isolation and Characterization of Mouse Bone Marrow Mesenchymal Stem Cells

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Six- to eight-week-old DBA1J mouse bone marrow cells were collected by flushing femurs and tibias with Dulbecco’s modified Eagle’s medium (Gibco, Carlsbad, CA, USA) containing 2 mM L-glutamine (Gibco), 1% antibiotics (penicillin (10 U/ml)–streptomycin (10 g/ml)) (Gibco) and 15% heat-inactivated fetal bovine serum (FBS, Gibco) with an endotoxin level ≤5 EU/ml and hemoglobin level ≤10 mg/dl (Gibco)52 (link). When cells reached around 80% confluency, the medium was aspirated and 3–5 ml trypsin-EDTA (Gibco) were added to each dish. The dishes were then incubated for ~5 min to allow cell detachment. Next, an equal volume of culture medium was added to inactivate trypsin. The marrow cell immunophenotypes were persistently positive for Sca-1 (D7; BioLegend, San Diego, CA, USA), CD44 (IM7; eBioscience, San Diego, Ca, USA), and CD29 (HMβ1-1; BioLegend), but negative for c-Kit (2B8; BioLegend), CD11b (M1/70; BD Pharmingen, San Diego, CA, USA), CD34 (MEC14.7; BioLegend), CD106 (429 (MVCAM.A); BD Pharmingen), CD45 (30-F11; BD Pharmingen), CD31 (MEC 13.3; BD Pharmingen), CD80 (16-10A1, BD Pharmingen), and CD86 (2331 (FUN-1), BD Pharmingen) after more than 10 passages (two months of culturing) (Supplementary Fig. 1).

Lim J.Y., Im K.I., Lee E.S., Kim N., Nam Y.S., Jeon Y.W, & Cho S.G. (2016). Enhanced immunoregulation of mesenchymal stem cells by IL-10-producing type 1 regulatory T cells in collagen-induced arthritis. Scientific Reports, 6, 26851.

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Multiparametric Flow Cytometry for PBMC Characterization

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Phenotypic characterization of the administrated PBMC subsets was analyzed by flow cytometry for the following fluorochrome-conjugated mAbs against the following surface antigens: CD11b (ICRF44), CD11c (B-ly6), CD206 (19.2), CD3 (SK7), CD31 (WM59), CD40 (5C3), CD44 (G44-26), CD45 (HI30), CD56 (HCD56), CD73 (AD2), CD79a (HM47), CD83 (HB15e), CD86 [2331 (FUN-1)], CD90 (5E10; BD Biosciences, San Jose, Calif., USA); CD34 (581; Biolegend, San Diego, Calif., USA); CD14 (Tük4), HLA-DR (Tü36; Invitrogen, UK). Data acquisition was performed using LSRFortessa (BD) equipped with a high-throughput system. The data were analyzed with FlowJo (TreeStar Inc.) software. SSC/FSC gates around single cell CD90 low CD79 + CD44 + CD73 + CD45 -CD34 -CD31 -CD11b -HLA-DR -populations were used to identify MSC populations. To identify PBMC populations, gating on SSC/FSC and single cells was performed first. CD3 - CD56 -identified CD83 + CD14 -cells as dendritic cells, CD83 + CD14 low as nondendritic (intermediate) and CD83 -CD14 + as monocytes. CD3 -CD56 + , CD3 + CD56 + and CD3 + CD56 -cells were identified as NK, NK-T and T cells, respectively (see fig. 2a).

Jungebluth P., Holzgraefe B., Lim M.L., Duru A.D., Lundin V., Heldring N., Wiklander O.P.B., Nordin J.Z., Chrobok M., Roderburg C., Sjöqvist S., Anderstam B., Beltrán Rodríguez A., Haag J.C., Gustafsson Y., Roddewig K.G., Jones P., Wood M.J.A., Luedde T., Teixeira A.I., Hermanson O., Winqvist O., Kalzén H., El Andaloussi S., Alici E, & Macchiarini P. (2015). Autologous Peripheral Blood Mononuclear Cells as Treatment in Refractory Acute Respiratory Distress Syndrome. Respiration; international review of thoracic diseases, 90(6).

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