Reh tumor cells were collected at 1000 rpm for 5 min. Cells were washed with 0.5% BSA-PBS once, and cell density was set to 5 × 105 cells/ml. Antibodies were diluted to the fixed concentrations (ML5 0.5 μg/ml and SN3 1.0 μg/ml) with 0.5% BSA-PBS. IMM47 antibody was diluted to 30 μg/ml followed by a 3-fold gradient dilution. 50 μl of cells and 50 μl of antibody dilution were added into each well of the 96-well U-shaped plates and incubated at 4 °C for 45 min. After incubation, wells were rinsed with 150 μl of the 0.5% BSA-PBS. A total of 100 μl of the diluted IMM47 were added and incubated at 4 °C for 45 min. After incubation, wells were rinsed with 150 μl of the 0.5% BSA-PBS solution. A total of 100 μl of the diluted anti-human IgG (Fc)-FITC (Sigma, Cat# F9512) was added and incubated at 4 °C for 45 min. After incubation, wells were rinsed with 150 μl of the 0.5% BSA-PBS was added. The cells were measured with flow cytometry. GraphPad Prism software was used to analyse the data, and the combination curves were drawn with four parameters.
Anti human igg fc fitc
Manufactured by Merck Group
The Anti-human IgG (Fc)-FITC is a fluorescently-labeled antibody that specifically binds to the Fc region of human immunoglobulin G (IgG) molecules. It is a laboratory reagent commonly used in various immunoassays and cell-based experiments to detect and quantify the presence of human IgG.
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Lab products found in correlation
6 protocols using anti human igg fc fitc
1
Flow Cytometric Analysis of Reh Cells
2
CD24 Glycosylation Modification Assay
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MCF-7, HCC1954 and Hela tumor cells were digested with trypsin and collected after centrifugation at 1000 rpm for 5 min. For different CD24 glycosylation modification assays, Reh cell suspensions with 2 μl of PNGase F (New England Bio Labs, Cat# P0710S), or Sialidase A (Prozyme, Cat# GK80040) were incubated at 37 °C for 24 h before the assay. Cells were collected at 1000 rpm for 5 min. Cells were washed with 0.5% BSA-PBS once, and cell density was set at 5 × 105 cells/ml. Antibodies were diluted to the corresponding concentration with three-fold gradient dilutions. About 50 μl of cells and 50 μl of antibody dilution were added into each well of the 96-well U-shaped plates and incubated at 4 °C for 45 min. After incubation, wells were rinsed with 150 μl of the 0.5% BSA-PBS solution. About 100 μl of the 1:500 diluted anti-human IgG (Fc)–FITC (Sigma, Cat# F9512) was added and incubated at 4 °C for 45 min. After incubation, wells were rinsed with 150 μl of the 0.5% BSA-PBS; and120 μl of the 0.5% BSA-PBS was added. The cells were measured with flow cytometry. GraphPad Prism software was used to analyse the data, and the combination curves were drawn with four parameters. The same method was used to verify the binding capacity of CD24-negative 293T cells.
3
Flow Cytometric Analysis of Reh Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Reh tumor cells were collected at 1000 rpm for 5 min. Cells were washed with 0.5% BSA-PBS once, and cell density was set to 5 × 105 cells/ml. Antibodies were diluted to the fixed concentrations (ML5 0.5 μg/ml and SN3 1.0 μg/ml) with 0.5% BSA-PBS. IMM47 antibody was diluted to 30 μg/ml followed by a 3-fold gradient dilution. 50 μl of cells and 50 μl of antibody dilution were added into each well of the 96-well U-shaped plates and incubated at 4 °C for 45 min. After incubation, wells were rinsed with 150 μl of the 0.5% BSA-PBS. A total of 100 μl of the diluted IMM47 were added and incubated at 4 °C for 45 min. After incubation, wells were rinsed with 150 μl of the 0.5% BSA-PBS solution. A total of 100 μl of the diluted anti-human IgG (Fc)-FITC (Sigma, Cat# F9512) was added and incubated at 4 °C for 45 min. After incubation, wells were rinsed with 150 μl of the 0.5% BSA-PBS was added. The cells were measured with flow cytometry. GraphPad Prism software was used to analyse the data, and the combination curves were drawn with four parameters.
4
CD24 Glycosylation Modification Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MCF-7, HCC1954 and Hela tumor cells were digested with trypsin and collected after centrifugation at 1000 rpm for 5 min. For different CD24 glycosylation modification assays, Reh cell suspensions with 2 μl of PNGase F (New England Bio Labs, Cat# P0710S), or Sialidase A (Prozyme, Cat# GK80040) were incubated at 37 °C for 24 h before the assay. Cells were collected at 1000 rpm for 5 min. Cells were washed with 0.5% BSA-PBS once, and cell density was set at 5 × 105 cells/ml. Antibodies were diluted to the corresponding concentration with three-fold gradient dilutions. About 50 μl of cells and 50 μl of antibody dilution were added into each well of the 96-well U-shaped plates and incubated at 4 °C for 45 min. After incubation, wells were rinsed with 150 μl of the 0.5% BSA-PBS solution. About 100 μl of the 1:500 diluted anti-human IgG (Fc)–FITC (Sigma, Cat# F9512) was added and incubated at 4 °C for 45 min. After incubation, wells were rinsed with 150 μl of the 0.5% BSA-PBS; and120 μl of the 0.5% BSA-PBS was added. The cells were measured with flow cytometry. GraphPad Prism software was used to analyse the data, and the combination curves were drawn with four parameters. The same method was used to verify the binding capacity of CD24-negative 293T cells.
5
CD24 Glycosylation Modification Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MCF-7, HCC1954 and Hela tumor cells were digested with trypsin and collected after centrifugation at 1000 rpm for 5 min. For different CD24 glycosylation modification assays, Reh cell suspensions with 2 μl of PNGase F (New England Bio Labs, Cat# P0710S), or Sialidase A (Prozyme, Cat# GK80040) were incubated at 37 °C for 24 h before the assay. Cells were collected at 1000 rpm for 5 min. Cells were washed with 0.5% BSA-PBS once, and cell density was set at 5 × 105 cells/ml. Antibodies were diluted to the corresponding concentration with three-fold gradient dilutions. About 50 μl of cells and 50 μl of antibody dilution were added into each well of the 96-well U-shaped plates and incubated at 4 °C for 45 min. After incubation, wells were rinsed with 150 μl of the 0.5% BSA-PBS solution. About 100 μl of the 1:500 diluted anti-human IgG (Fc)–FITC (Sigma, Cat# F9512) was added and incubated at 4 °C for 45 min. After incubation, wells were rinsed with 150 μl of the 0.5% BSA-PBS; and120 μl of the 0.5% BSA-PBS was added. The cells were measured with flow cytometry. GraphPad Prism software was used to analyse the data, and the combination curves were drawn with four parameters. The same method was used to verify the binding capacity of CD24-negative 293T cells.
6
Flow Cytometric Analysis of Reh Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Reh tumor cells were collected at 1000 rpm for 5 min. Cells were washed with 0.5% BSA-PBS once, and cell density was set to 5 × 105 cells/ml. Antibodies were diluted to the fixed concentrations (ML5 0.5 μg/ml and SN3 1.0 μg/ml) with 0.5% BSA-PBS. IMM47 antibody was diluted to 30 μg/ml followed by a 3-fold gradient dilution. 50 μl of cells and 50 μl of antibody dilution were added into each well of the 96-well U-shaped plates and incubated at 4 °C for 45 min. After incubation, wells were rinsed with 150 μl of the 0.5% BSA-PBS. A total of 100 μl of the diluted IMM47 were added and incubated at 4 °C for 45 min. After incubation, wells were rinsed with 150 μl of the 0.5% BSA-PBS solution. A total of 100 μl of the diluted anti-human IgG (Fc)-FITC (Sigma, Cat# F9512) was added and incubated at 4 °C for 45 min. After incubation, wells were rinsed with 150 μl of the 0.5% BSA-PBS was added. The cells were measured with flow cytometry. GraphPad Prism software was used to analyse the data, and the combination curves were drawn with four parameters.
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