Cells were detached with trypsin-EDTA (ThermoFisher, Waltham, MA), washed, and resuspended in cold PBS buffer containing 5% BSA. One million cells were co-incubated with GPC3 mAbs (1 μg/ml, or as indicated in specific experiments). The antibody binding was detected by Allophycocyanin (APC)-conjugated goat anti-human IgG (Jackson ImmunoResearch Inc, West Grove, PA). The fluorescence intensity (Geo. Mean) was measured by using BD FACS Aria. To confirm the cell-binding activity of immunotoxins, one million G1 cells were incubated with 10 μg/ml immunotoxin on ice for 1 hour. The binding of immunotoxin was detected by APC-conjugated mouse anti-His antibody (BioLegend).
Apc conjugated mouse anti his antibody
Manufactured by BioLegend
The APC-conjugated mouse anti-His antibody is a fluorescently labeled antibody that specifically binds to the histidine (His) tag on recombinant proteins. This antibody can be used in various applications, such as flow cytometry, to detect and quantify His-tagged proteins.
Automatically generated - may contain errors
3 protocols using apc conjugated mouse anti his antibody
1
GPC3 Antibody-Binding Assay
2
SARS-CoV-2 Spike Protein Binding to THP1 Cells
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To evaluate SARS-CoV-2 S binding to various THP1 monocytes expressing different surface receptors, approximately 0.25x106 cells from parental THP1 or those expressing wt CD169, mutant CD169 (R116A), ACE2, or both wt CD169 and ACE2 were incubated for 30 min at 4°C with 2 μg of spike glycoprotein (stabilized) from Wuhan-Hu-1 SARS-CoV-2 containing a C-terminal Histidine Tag, recombinant from HEK293F cells (BEI resources, #NR-52397). This is followed by secondary staining for 30 min at 4°C with APC-conjugated mouse anti-His antibody (BioLegend, #362605, 1:50) or isotype control. Cells were fixed with 4% PFA (Boston Bioproducts) for 30 min, and analyzed with BD LSRII (BD). Data analysis was performed using FlowJo software (FlowJo).
3
Evaluating SARS-CoV-2 Spike Binding
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To evaluate SARS-CoV-2 S binding to various THP1 monocytes expressing different surface receptors, approximately 0.25×106 cells from parental THP1 or those expressing wt CD169, mutant CD169 (R116A), ACE2, or both wt CD169 and ACE2 were incubated for 30 min at 4 °C with 2 μg of spike glycoprotein (stabilized) from Wuhan-Hu-1 SARS-CoV-2 containing a C-terminal Histidine Tag, recombinant from HEK293F cells (BEI resources, #NR-52397). This is followed by secondary staining for 30 min at 4°C with APC-conjugated mouse anti-His antibody (BioLegend, #362605, 1:50) or isotype control. Cells were fixed with 4% PFA (Boston Bioproducts) for 30 min, and analyzed with BD LSRII (BD). Data analysis was performed using FlowJo software (FlowJo).
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