To detect surface B7-H4, CD11b or CD133 expression, cells were incubated on ice for 30min with appropriate antibodies or isotype controls (PE-B7-H4, PE-mouse IgG1, APC-CD11b, APC-CD133, APC-mouse IgG1, all from eBioscience, 1:5), washed twice with FACS buffer (PBS containing 0.1% NaN3 and 5% fetal bovine serum), and resuspended in 0.5 ml 1% formalin/PBS. To analyze intracellular B7-H4, cells were pre-treated with Fix and Perm cell permeabilization reagents (Caltag Laboratories) according to the manufacturer's instructions. Assays of immune function, cell proliferation, cell cycle, apoptosis, cytokines and cytotoxicity were done under standard methods as previously described. All analyses were performed on a FACS calibur system (Becton Dickinson Immunocytometry Systems).
Apc mouse igg1
Manufactured by Thermo Fisher Scientific
Sourced in United States
The APC-mouse IgG1 is a fluorescently-labeled antibody used for flow cytometry analysis. It binds to mouse immunoglobulin G1 (IgG1) antibodies. The APC (allophycocyanin) fluorescent dye is conjugated to the mouse IgG1 antibody, enabling detection and quantification of target cells or molecules.
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Lab products found in correlation
Mouse igg1 pe, by Thermo Fisher Scientific (3 mentions)
Facscalibur system, by BD (1 mentions)
Cd11b apc, by Thermo Fisher Scientific (1 mentions)
Pe b7 h4, by Thermo Fisher Scientific (1 mentions)
Anti human cd4 fitc, by BD (1 mentions)
Pe mouse igg1, by BD (1 mentions)
Mouse igg1 fitc, by BD (1 mentions)
Anti human cd8 fitc, by BD (1 mentions)
Anti human cd14 fitc, by BD (1 mentions)
Fitc anti human cd56, by BD (1 mentions)
Mouse igg1 fitc, by Thermo Fisher Scientific (1 mentions)
Phospho smad2 3, by Cell Signaling Technology (1 mentions)
Anti human cd4 pe, by BD (1 mentions)
Anti human foxp3 apc, by Thermo Fisher Scientific (1 mentions)
Smad2 3 d7g7, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Kaluza software, by Beckman Coulter (1 mentions)
Gallios flow cytometer, by Beckman Coulter (1 mentions)
Cytofix cytoperm kit, by BD (1 mentions)
4 protocols using apc mouse igg1
1
Detecting Surface and Intracellular Markers
2
Flow Cytometric Analysis of IL-17RA Expression
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SV40 fibroblasts were plated in 24-well plates at a density of 50,000 cells/well, in 0.5 ml of 10% FBS in DMEM per well. The plates were incubated for 24 h. The cells were then subjected to flow cytometry. SV40-transformed fibroblasts or PBMCs were labeled with PE-mouse IgG1 (eBioscience), APC-mouse IgG1 (eBioscience), Alexa Fluor 647 mouse IgG1, κ isotype ctrl (FC) antibody (Sony Biotechnology, Inc.), VioGreen-mouse IgG2a (Miltenyi Biotec), VioBlue-mouse IgG2a (Miltenyi Biotec), FITC-mouse IgG1 (BD), FITC-mouse IgG2b (BD), and PE-mouse IgG1 (BD) antibodies or Alexa Fluor 647–mouse IgG1 (eBioscience) isotype antibodies as a control. PE-anti–human IL-17RA (clone J10MBS; eBioscience), APC-anti–human IL-17RA (clone 424LTS; eBioscience), Alexa Fluor 647 anti–human CD217 (IL-17RA) antibody (Sony Biotechnology, Inc.), VioGreen-anti–human CD14 (Miltenyi Biotec), VioBlue-anti–human CD3 (Miltenyi Biotec), FITC-anti–human CD4 (BD), FITC-anti–human CD8 (BD), FITC-anti–human CD14 (BD), FITC-anti–human CD19 (BD), and FITC-anti–human CD56 (BD) specific antibodies were used according to the manufacturers’ instructions, with the results analyzed by flow cytometry on a FACSCanto II system.
3
Multiparameter Flow Cytometry Immunophenotyping
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Monoclonal antibodies against CD4-FITC (RPA-T4), CD11b-APC (ICRF44), CD45RA-APC (5H9), and FoxP3-PE (259D/C7) were purchased from Becton Dickinson (Franklin Lakes, NJ, USA). LAP-PE (clone 27,232) was purchased from R&D Systems (Minneapolis, MN, USA). Mouse IgG1-FITC, mouse IgG1-PE, and mouse IgG1-APC were purchased from eBioscience (San Diego, CA, USA) and were used as isotype-matched negative controls. Human Fc receptor blocker was purchased from Becton Dickinson. Ethidium monoazide bromide was purchased from Molecular Probes, Inc. (Eugene, OR, USA).
4
Regulatory T Cell Expansion Assay
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Human PBMCs (ALLCELLS) were stained with anti-CD3 and Glycophorin A to enumerate T cells. Cells were cultured with rhIL-2 (100 IU ml−1) and anti-CD3/anti-CD28-coated Dynabeads (Life Technologies) at a ratio of 1 : 3 (cell : bead) in the presence or absence of 2.5 ng ml−1 rhTGF-β1 with or without either a-CTLA4-TGFβRII or a-CTLA-4 antibody (5 μg ml−1). Following culture for 24–48 h, cells were lysed and subjected to immunoblot analyses with the following primary antibodies: FOXP3, SMAD-2/3 (D7G7), or phospho-SMAD-2/3, and β-actin (Cell Signaling Technologies). On day 5, anti-CD3/anti-CD28 beads were magnetically removed and the number of Tregs (CD4+/CD25high/CD127low/FOXP3+) were enumerated by flow cytometry. Cells were stained extracellularly with anti-human CD4-PE, anti-human CD25-PE-CYTM5, and anti-human CD127-FITC antibodies (BD Biosciences). The cells were then permeabilized (BD Cytofix/Cytoperm Kit) and stained intracellularly with anti-human FOXP3-APC or the corresponding isotype control mouse IgG1-APC (eBioscience). The stained cells were washed twice with FACS buffer, run on the Gallios Flow Cytometer, and analyzed utilizing Kaluza Software (Beckman Coulter).
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