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In addition to the broad range of flow cytometry validated antibodies we offer for immunology, cancer and veterinary science, we also have supporting reagents that will optimize staining and sample preparation. To help you use these reagents, we have created a free poster, available to download or order, which takes you through the important stages you should follow to design a successful flow cytometry experiment.
Leucoperm™– cell permeabilization reagent for intracellular staining
Leucoperm reagent allows intracellular antigen analysis with the same ease as surface antigens. Structures such as cytoplasmic or nuclear enzymes, oncoproteins, cytokines and cytoplasmic localizations of established membrane molecules can be stained with antibodies. Leucoperm allows you to fix and permeabilize cells in suspension whilst retaining their morphology.
Fc receptors are found on monocytes, macrophages, dendritic cells and B cells. As the name suggests they bind antibodies via their constant Fc domain rather than the antigen specific Fab domain. This can lead to false positives. In order to prevent this type of binding, blocking agents have been developed to ensure that only antigen specific binding is observed.
Mouse Seroblock – for blocking Fc receptors on mouse cells
Human Seroblock – for blocking Fc receptors on human cells
Fig.1. Staining of THP-1 cells with FITC labeled mouse anti-human CD11a in blue (MCA1848) or mouse IgG2a negative control:FITC red (MCA929F) in the absence (a) and presence (b) of human SeroBlock (BUF070A).
Red cell lysing buffer – Erythrolyse is available as a 10x solution stable for 1 year at room temperature. It is suitable for the lysing of red blood cells in human, rat and mouse blood.
Viability dyes – we have easy-to-use cell viability assays in a range of excitation and emission wavelengths, suitable for most multi-color flow cytometry. These include DNA binding dyes such as PI, 7-AAD and DAPI and our protein binding VivaFixTM dyes which are fixable to allow analysis at a later date.
Proliferation dyes – our cell proliferation assays include dyes, TUNEL assay kits and BrdU antibodies to give you flexibility in your experiments. They are also available in a range of excitation and emission wavelengths for your convenience.
Isotype controls – determine the level of background surface staining due to non-specific antibody or fluorophore binding. We have isotype controls for mouse, rat, hamster and goat antibodies.
When performing multicolor flow cytometry, there are several important steps you must consider. Careful sample preparation, Fc blocking, inclusion of a live/dead dye, doublet exclusion and using the appropriate controls are all important. For more information watch our flow cytometry webinar Optimize your Flow Cytometry or visit our application resources page.
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