The lung cancer cells were incubated with mouse anti-MICA, anti-MICB, anti-ULBP1-3, and anti-HLA-ABC monoclonal antibodies (mAbs) (R&D systems, Minneapolis, MN, USA) to determine the expression of NKG2D ligands and MHC class I molecules in lung cancer cells. Specific mAbs against NKG2D ligands and MHC class I molecules or the corresponding isotype controls were used at 1 μg in 100 μL cell suspensions. These were followed by the addition of goat anti-mouse PE-conjugated antibody (BD Phamingen Inc., San Diego, CA, USA). The analysis was performed using BD FACSCANTO II (Becton Dickinson and Company, Franklin Lakes, NJ, USA) and FlowJo software (BD Biosciences, Franklin Lakes, NJ, USA). Surface expression levels were measured as mean fluorescence intensity (MFI).
Anti ulbp1 3
Manufactured by R&D Systems
Sourced in United States
Anti-ULBP1-3 is a monoclonal antibody that binds to ULBP1, ULBP2, and ULBP3 proteins. ULBPs are ligands for the NKG2D receptor, which is expressed on natural killer cells, CD8+ T cells, and other immune cells. This antibody can be used to detect the expression of ULBP1, ULBP2, and ULBP3 in various research applications.
Automatically generated - may contain errors
5 protocols using anti ulbp1 3
1
Characterizing NKG2D Ligands in Lung Cancer
2
Quantifying NKG2D Ligand Expression
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To determine the surface expression of the NKG2D ligands on cancer cells, the cells were incubated with mouse anti-MICA, anti-MICB, anti-ULBP1–3 (R&D systems, Minneapolis, MN, USA), which were NKG2D ligand–specific monoclonal antibodies (mAbs), and the corresponding isotype controls at 10 μg/m, followed by incubation with goat anti-mouse-PE conjugated (BD Phamingen Inc., San Diego, CA., USA). The analysis was performed on a FACS Caliber® (Becton Dickinson, Mountain View, CA., USA) using CellQuest software (Becton Dickinson), and the cell surface expression was quantified by the value of the mean fluorescence intensities (MFIs) obtained with the specific mAbs.
3
Quantifying NKG2D Ligands on Cancer Cells
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To determine the surface expression of the NKG2D ligands on cancer cells, the cells were incubated with mouse anti-MICA, anti-MICB, and anti-ULBP1-3 (R&D Systems), which were NKG2D ligand-specific monoclonal antibodies (mAbs), and the corresponding isotype controls at 10 µg/ml, followed by incubation with the goat anti-mouse-PE conjugated (BD Pharmingen Inc.). The analysis was performed on the FACSCalibur® system using the CellQuest software (both from Becton-Dickinson), and the cell surface expression was quantified from the value of the mean fluorescence intensities (MFI) obtained with the specific mAbs.
4
Quantifying NKG2D Ligand Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To determine the surface expressions of NKG2D ligands on cancer cells, the cells were incubated with mouse anti-MICA, anti-MICB, anti-ULBP1-3 (R&D systems, Minneapolis, MN, USA), anti-HLA-ABC (Clone W6/32, Serotec, Oxford, UK) or the corresponding isotype controls at 10 μg/ml and then incubated with goat anti-mouse-PE conjugated (BD Pharmingen Inc., San Diego, CA., USA). The analysis was performed on a FACS Sort® (Becton Dickinson, Mountain View, CA., USA) using Cell Quest software (Becton Dickinson), and cell surface expressions were quantified using mean fluorescence intensities (MFIs). Relative expression ratios were calculated by dividing treated sample MFI by untreated sample MFI without subtracting the MFI of the appropriate isotype control.
5
Quantifying NKG2D Ligands on Cancer Cells
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To determine the surface NKG2DLs on cancer cells, the cells were incubated with mouse anti-MICA, anti-MICB, anti-ULBP1–3 (R&D systems, Minneapolis, MN, USA), which were NKG2D ligand-specific monoclonal antibodies (mAbs) or the corresponding isotype controls at 1 μg/100 μl followed by incubation with PE goat anti-mouse Ig (BD Phamingen Inc., San Diego, CA, USA). The analysis was performed on the BD FACSCANTO II (Becton Dickinson and Company, Franklin Lakes, NJ, USA) using FlowJo software (BD Biosciences, Franklin Lakes, NJ, USA) and the cell surface expression was quantified by the value of the mean fluorescence intensities (MFI) obtained with the specific mAbs [48 (link)].
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