T-cytotoxic (CD45+CD3+CD8+), T-helper (CD45+CD3+CD4+), and B (CD45+CD3-CD20+) lymphocytes were separated using FACS. PBMCs (1 × 106 cells/mL) were stained with FITC anti-human CD3 Antibody (300312, BioLegend, San Diego, CA, USA), APC anti-human CD4 Antibody (357404, BioLegend, San Diego, CA, USA), PE anti-human CD8 Antibody (2323530, Sony, San Jose, CA, USA), APC/Cy7 anti-human CD14 Antibody (2109100, Sony, San Jose, CA, USA), and Brilliant Violet 421™ Anti-human CD20 Antibody (2111650, Sony, San Jose, CA, USA) following the manufacturer’s recommendations. In 96-well plates, 1.5 × 105 cells per well were seeded. CIMVs-MSCs (10 μg per well) were added 24 h after the sorting.
Apc anti human cd4 antibody
Manufactured by BioLegend
Sourced in United States
The APC anti-human CD4 Antibody is a fluorochrome-conjugated monoclonal antibody that binds to the CD4 surface antigen expressed on T helper cells. It is designed for use in flow cytometry analysis to identify and quantify CD4-positive cells within a sample.
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Lab products found in correlation
3 protocols using apc anti human cd4 antibody
1
Lymphocyte Subtyping and CIMV-MSC Interaction
2
Transduction and Flow Cytometry Analysis
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HT29 cells were transduced with virus for each of the guide+dCas12a-TAD-containing vectors separately; 5 days after transduction, cells were visualized by flow cytometry on a CytoFLEX S Sampler. To prepare samples for visualization, cells were stained with APC anti-human CD4 Antibody (Biolegend, 357408), diluted 1:100 for 20–30 min on ice.
Cells were washed with PBS two times to remove residual antibody and were resuspended in flow buffer (PBS, 2% FBS, 5 μM EDTA). CD4 signal was measured in the APC-A channel and VexGFP signal was measured in the K0525-A channel. Flow cytometry data were analyzed using FlowJo (v10.8.1). Cells were gated for VexGFP expression and APC gates were drawn such that ∼1% of cells score as APC-positive in the control condition (stained parental cells).
Cells were washed with PBS two times to remove residual antibody and were resuspended in flow buffer (PBS, 2% FBS, 5 μM EDTA). CD4 signal was measured in the APC-A channel and VexGFP signal was measured in the K0525-A channel. Flow cytometry data were analyzed using FlowJo (v10.8.1). Cells were gated for VexGFP expression and APC gates were drawn such that ∼1% of cells score as APC-positive in the control condition (stained parental cells).
3
Genetic Manipulation and Flow Cytometry Analysis
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HT29, MelJuSo, HCC2429 and/or A375 cells were transduced with virus for each of the dCas12a-containing vectors separately; 2 days after transduction, cells were selected with blasticidin for 14 days. Blasticidin was removed for one passage and cells were subsequently transduced with virus for guide+nanobody-TAD-containing vectors. 2 days after transduction, cells were selected with puromycin for 5 days. Following selection, cells were visualized by flow cytometry on a CytoFLEX S Sampler at varying time points. To prepare samples for visualization, cells were stained with a fluorophore-conjugated antibody targeting the respective cell surface marker gene, diluted 1:100 for 20–30 min on ice.
CD4: APC anti-human CD4 antibody (Biolegend, 357408)
CD26 (DPP4): FITC anti-human CD26 antibody (Biolegend, 302704)
CD274: APC anti-human CD274 antibody (Biolegend, 329708)
CD97 (ADGRE5): FITC anti-human CD97 antibody (Biolegend, 336306)
Cells were washed with PBS two times to remove residual antibody and were resuspended in flow buffer (PBS, 2% FBS, 5 μM EDTA). Fluorophore signal was measured in the respective channel (APC-A or FITC-A). Flow cytometry data were analyzed using FlowJo (v10.8.1). Gates were set such that ∼1% of cells score as APC-positive or FITC-positive in the control condition (stained parental cells).
CD4: APC anti-human CD4 antibody (Biolegend, 357408)
CD26 (DPP4): FITC anti-human CD26 antibody (Biolegend, 302704)
CD274: APC anti-human CD274 antibody (Biolegend, 329708)
CD97 (ADGRE5): FITC anti-human CD97 antibody (Biolegend, 336306)
Cells were washed with PBS two times to remove residual antibody and were resuspended in flow buffer (PBS, 2% FBS, 5 μM EDTA). Fluorophore signal was measured in the respective channel (APC-A or FITC-A). Flow cytometry data were analyzed using FlowJo (v10.8.1). Gates were set such that ∼1% of cells score as APC-positive or FITC-positive in the control condition (stained parental cells).
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