General notesThis product is a mixture of the same clone separately bearing both FITC and PE labels.
This product contains sodium azide, which under acid conditions yields hydrazoic acid, a toxic compound.
Azide compounds should be diluted with running water before being discarded to avoid deposits in lead or copper plumbing where explosive conditions may develop.
This antibody control duo is a direct immunofluorescence reagent used as an isotype (negative) control for flow cytometric immunophenotyping of erythrocyte-lysed whole blood with monoclonal antibodies.
Storage instructionsShipped at 4°C. Store at +4°C.
Storage bufferPBS with 0.5% BSA and 0.1% sodium azide
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Isotype control notesThis antibody control duo is a direct immunofluorescence reagent used as an isotype (negative) control for flow cytometric immunophenotyping of erythrocyte-lysed whole blood with monoclonal antibodies.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
This antibody control duo is used to determine the unstained lymphocyte population and to determine any non-antigen-specific antibody binding on mononuclear cells separated by density gradient or on lysed whole blood. IgG1 (FITC + PE) immunofluorescence analysis can be performed on a flow cytometerequipped with an excitation source of 488nm and fitted with logarithmic amplifiers. Scatter gates are set on the lymphocyte fraction. Proper electronic compensation and filter selections are necessary for three-color analysis. 10µl ofIgG1 (FITC + PE) is sufficient for labelling of 1x106 cells.
Mouse IgG1monoclonal [HybIgG1] (FITC + Phycoerythrin) - isotype control images
ICC/IF image of ab2185 stained MCF7 cells. The cells were 100% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2185, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
References for Mouse IgG1monoclonal [HybIgG1] (FITC + Phycoerythrin) - isotype control (ab1285)
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