1 × 106 LS174T cells were detached with trypsin, centrifuged and incubated with an EGFR-specific antibody (1:100; monoclonal mouse IgG1 - Dako, Glostrup, Denmark) or with an IgG-anti-mouse antibody (BD Bioscience, Franklin Lakes, USA) as negative control in FACS buffer (PBS with 10% FBS) for 1 h on ice. Cells were washed with FACS buffer and incubated with an AlexaFluor 488 labeled goat anti-mouse secondary antibody (1:400 - Invitrogen, Langenselbold, Germany) for 1 h on ice. After washing, cells were resuspended in FACS buffer and flow cytometry analysis was performed on a BD Accuri C6 flow cytometer (BD Bioscience, Franklin Lakes, USA). Cells were gated by forward/sideward scatter and pulse width for exclusion of doublets. PI (propidium iodide, Sigma-Aldrich) was used for discrimination between viable and dead cells.
Monoclonal mouse igg1
Manufactured by Agilent Technologies
Sourced in Denmark
Monoclonal mouse IgG1 is a laboratory reagent used in various immunological and biochemical applications. It is a purified antibody produced by a single clone of mouse B cells. The core function of this product is to serve as a specific and consistent binding reagent for detecting or quantifying target analytes in samples.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488 labeled goat anti mouse secondary antibody, by Thermo Fisher Scientific (2 mentions)
Igg anti mouse antibody, by BD (2 mentions)
Accuri c6 flow cytometer, by BD (2 mentions)
Phosphate buffered saline (pbs), by Merck Group (2 mentions)
Pi propidium iodide, by Merck Group (1 mentions)
Monoclonal mouse igg1, by R&D Systems (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Trypsin edta, by Merck Group (1 mentions)
2 protocols using monoclonal mouse igg1
1
EGFR Expression Analysis in LS174T Cells
2
EGFR and cMET/HGFR Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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In total, 0.8-1 × 10 6 cells were detached with trypsin/EDTA (Sigma-Aldrich), washed with phosphate buffered saline (PBS; Sigma-Aldrich), supplemented with 10% FBS (Sigma-Aldrich) and incubated with an EGFR-specific antibody (1:200; monoclonal mouse IgG1, Dako, Glostrup, Denmark) or with an antibody that detects human cMET/HGFR (1:200; monoclonal mouse IgG1, R&D Systems, Minneapolis, Minnesota, USA) or with an IgG-antimouse antibody (1:200; BD Biosciences, Franklin Lakes, New Jersey, USA) as negative control for 1 h on ice. Then, cells were washed with PBS with 10% FBS and incubated with an AlexaFluor 488 labeled goat anti-mouse secondary antibody (1:400; Invitrogen, Langenselbold, Germany) for 1 h on ice. Cells were washed and resuspended in PBS with 10% FBS for analysis, which was performed on a BD Accuri C6 flow cytometer (BD Biosciences). Cells were gated by forward/sideward scatter and pulse width for exclusion of doublets. PI (propidium iodide; Sigma-Aldrich) was used for discrimination between viable and dead cells.
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