Antibodies for Immune Cell Therapy
Find highly specific monoclonal and recombinant monoclonal antibodies for activation, expansion, and characterization of T cell and NK cell therapies.
LlaMAbodiesTM: Single Domain Antibodies
- Smaller size
- Longer CDR3 region
- Superior stability
We offer engineered VHH single domain, bivalent and IgG-scaffold antibodies.
Quality Control and Standardization Beads
Set yourself up for successful flow cytometry experiments by using our Calibration, Compensation, Cell Counting, and Equivalent Reference Fluorochromes (ERF) beads.
BSA and Azide Free Antibodies
Over 2000 antibodies available in CyTOF-ready format lyophilized with Trehalose or resuspended in sterile PBS.
Biosimilar and Anti-Idiotype Antibodies
Attack cancer with an arsenal of powerful research tools including biosimilar and anti-idiotype antibodies.
Flow Cytometry Antibodies
Find our full selection of flow cytometry validated primary antibodies, including R&D Systems clones, at Novus Biologicals. Explore our broad selection of fluorescently conjugated antibodies, including Alexa Fluor® 488, Alexa Fluor® 647, PE, PE-Cy7TM, and more!
Tools for Cell Selection
- Our T Cell Enrichment Columns are designed to use high-affinity negative selection to enrich and purify T cells and specific T cell subpopulations.
- MagCellectTM Cell Selection Kits isolate and enrich cell populations through magnetic, bead-based positive or negative selection without the need for specialized columns.
Flow Cytometry Handbook
Our Flow Cytometry Handbook is the must have resource for flow cytometry researchers. Download now to find step-by-step protocols and troubleshooting tips.
Flow Cytometry Webinars
Our two-part flow cytometry webinar series:
- The first, Demystifying multi-parameter flow cytometry, learn flow basics like spectral overlap, compensation, optimal fluorochrome choice, and data analysis.
- The second, Turning Flow Cytometry Upside Down and Inside Out, will apply the basics to more complex flow topics including cell proliferation, dead cell exclusion, and intracellular staining.
This new eBook provides an overview of several biological and manufacturing challenges facing the development and production of T cell therapies. It follows with examples of how Bio-Techne products and services can address these challenges at each stage of the process.
Background Information, FAQs, Resources and Products
Flow cytometry is a high throughput, single-cell analysis technique that uses lasers to rapidly activate fluorescently-labeled conjugated antibodies on cells in a suspended solution. It is a powerful tool as it can measure many parameters on single cells, as well as very quickly collect information from millions of cells. In flow cytometry, single-cell suspensions are passed through single or multiple lasers, which causes the light to scatter. Cell populations can be sorted based on light scatter, where forward scatter provides cell size analysis and side scatter determines cell granularity. Flow cytometry can provide information about cell size and volume, evaluate the purity of isolated cell subpopulations, characterize different cell types in a heterogeneous population, while providing a technique that can examine the expression of both cell surface and intracellular proteins.
Fluorophore compounds, such as fluorescently-labeled antibodies or propidium iodide, are commonly used in flow cytometry to detect proteins of interest. Using fluorophores with nonoverlapping maximum emission spectra enables simultaneous individual detection of fluorescently stained cells. Isotype control antibodies are used as negative controls to validate a flow cytometry experiment. Multiple research areas including cancer biology, immunology, infectious disease monitoring, virology, and molecular biology utilize flow cytometry.
What is FACS?
Fluorescence-activated cell sorting (FACS) is a specialized type of flow cytometry. It provides a method for sorting a heterogeneous mixture of biological cells into separate containers, one cell at a time. Cell sorting is based upon the specific light scattering and fluorescent characteristics of each cell.
What is the minimum number of cells needed for flow cytometry?
The number of cells needed for a particular experiment is dependent on your needs. You would need fewer cells if you are probing for a robustly expressed antigen compared to a sparsely expressed protein. Additionally, you would need more cells for multicolor flow cytometry versus single color staining. However, we recommend starting with 1 × 106 cells per 100 µl of staining buffer per tube.
What controls are used in flow cytometry?
Flow cytometry controls can be split into the following categories.
- Instrument Controls: Calibration particles or fluorescent beads from the instrument’s manufacturer should be run prior to use to ensure the instrument is performing properly.
- Compensation Controls: When performing multicolor flow cytometry, the emission spectra from the different fluorophores used can overlap and can spread into multiple channels. The process of correcting for spillover is known as compensation. For compensation, a single stained sample is needed from each fluorochrome in the experiment. These can be set up using cells, compensation beads, or antibody capture beads.
- Gating Controls: One of the most critical aspects of analyzing flow cytometry data is properly setting gates. The Fluorescence Minus One (FMO) control is necessary for interpreting flow data and for setting gates. Since FMO controls provide a reliable assessment of background in a multicolor experiment, they can accurately discriminate between negative and positive cell populations.
- Cell Viability Controls: Dead cells can produce false positives because they autofluoresce and display an increased affinity for non-specific antibody binding. As a result, these dead cells need to be removed from data analysis. Several fluorescent dyes, such as 7-AAD, calcein AM, and propidium iodide, can be used to distinguish dead cells from live cells.
- Unstained Cells Controls: Several cell components, such as flavin coenzymes and NADPH, are endogenous fluorophores that exhibit autofluorescence. Running a sample of unstained cells through the flow cytometry will allow you to determine the amount of background fluorescence due to autofluorescence so that you can gate appropriately.
- Isotype Controls: Isotype controls are used to assess non-specific binding due to antibody class. An isotype control is an antibody that lacks specificity to the target of interest, but matches the class, type, and fluorescent conjugate of the primary antibody.
Supplement your Flow Cytometry Experiments with Milo™
If you need to analyze poorly expressed intracellular targets, or are using samples with low cell numbers, Single-Cell Westerns using Milo from ProteinSimple, a Bio-Techne brand, can complement your flow cytometry experiments.