Isotype controls are primarily used as negative controls in flow cytometry but they can also be used for immunohistochemistry. They are used to approximate the non-specific target primary antibody binding due to protein-protein interactions, binding to Fc receptors on target cells, non-specific protein-protein interactions, and cell autofluorescence. In order to most accurately decrease the non-specific background signal, the isotype control used should be selected to match as closely as possible to the primary antibody with respect to species, isotype (heavy and light chains), and conjugation type. Isotype controls are available for almost all immunoglobulin subclasses. The Rabbit IgG isotype control antibody has been raised against normal rabbit IgG isolated using fractionation and DEAE chromatography. Rabbit IgG isotype control antibody was used by Pallet et al in their comprehensive characterization of extracellular membrane vesicles, where they examined their components using the autophagic human endothelial cell as their model system1.
Flow Cytometry: Rabbit IgG Isotype Control
They used the Rabbit IgG isotype control antibody in flow cytometry studies that were then integrated into biochemical, ultrastructural, morphological, and proteome studies where they determined that these extracellular bodies do not contain nucleus components and differ significantly from classic apoptotic bodies. Some recent and interesting ChIP studies out of Johns Hopkins used the Rabbit IgG isotype control antibody to examine tumor microenvironments, and found that intratumoral hypoxia triggers RhoA-ROCK1 signaling pathways in breast cancer2. Mazzoni’s group at the Columbia University Medical Center performed Rabbit IgG isotype control antibody immunoprecipitations in their transcriptional analyses of the Ngn2, ISl1, and Lhx3-mediated spinal motor neuron identity programming in differentiating mouse embryonic stem (ES) cells3.
Novus Biologicals offers Rabbit IgG reagents for your research needs including:
