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Products for Detecting and Defining Cells
Flow Cytometry Antibodies
Flow Cytometry Antibodies
We offer conjugated and unconjugated primary antibodies for flow cytometry. Choose from 8,000+ flow cytometry-validated antibodies and 20 different fluorescent conjugates.
Conjugated Secondary Antibodies
Conjugated Secondary Antibodies
Indirect detection is more sensitive and vital for identification of low abundance antigens and rare epitopes in flow cytometry. For targets where low expression is observed, we have conjugated secondary antibodies for use with your primary antibody for the target of interest using indirect detection.
Isotype Control Antibodies
Isotype Control Antibodies
Isotype control antibodies are used as negative controls in flow cytometry. We offer a collection of isotype controls, both conjugated and unconjugated, for multiple isoforms and species.
CyTOF-Ready Antibodies
CyTOF-Ready Antibodies
We offer a selection of antibodies lyophilized in a format (azide free, carrier free) that makes them ready for conjugation to heavy metals for use in CyTOF/Mass Cytometry
Multicolor Flow Cytometry Kits
Multicolor Flow Cytometry Kits
Our Multicolor Flow Cytometry Kits contain fluorochrome-conjugated antibodies for simultaneous detection of up to 4 different target molecules on a single-cell.
Supplemental Reagents
Supplemental Reagents
We have a variety of flow cytometry buffers for cell permeabilization, fixation and staining.
Flow Cytometry Compatible Dyes from Tocris
Explore the range of flow cytometry compatible dyes from Tocris, a Bio-Techne brand. Dyes are grouped according to nearest laser lines. Conjugation protocols available.
The Novus Panel Builder Tool
The Novus Panel Builder Tool
Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool from Novus Biologicals, a Bio-Techne brand.
Interactive Cell Marker Tool
Interactive Cell Marker Tool
Learn more about cell surface and intracellular markers from R&D Systems, a Bio-Techne brand.
Custom Antibody Services
If you can’t find the antibody that you need on our website, contact us. Custom antibody production is a complicated process. Leverage the experience and skill of our antibody development scientists for your custom antibody project.
Bio-Techne's Flow Cytometry Handbook
Our Flow Cytometry Handbook is the must have resource for researchers doing flow cytometry. Download now to see the step-by-step protocols and troubleshooting tips.
Products for Selecting Cells
T Cell Enrichment Columns
T Cell Enrichment Columns
Our T Cell Enrichment Columns are designed to use high-affinity negative selection to enrich and purify T cells and specific T cell subpopulations.
MagCellect™ Cell Selection Kits
MagCellect™ Cell Selection Kits
We offer kits for enriching cell populations through magnetic bead-based positive or negative selection so specialized columns are not needed.
HLDA Antibodies
R&D Systems™ brand antibodies were used by HLDA to establish CD designations. View our trusted HLDA antibodies.
Products for Differentiating and Expanding Cells
Cell Culture Reagents
Cell Culture Reagents
Bio-Techne offers a comprehensive portfolio of reagents for cell culture including classic media, balanced salt solutions, FBS, supplements, and basement membrane extracts.
Cell Culture Media Supplements
Cell Culture Media Supplements
We offer validated media supplements for the culture of cell lines and primary cells, such as neurons and stem cells.
CellXVivo™ Differentiation and Expansion Kits
CellXVivo™ Differentiation and Expansion Kits
We have bundled together reagents, including optimized cytokine combinations, for differentiating and expanding immune cells, such as T cells, B cells, macrophages, and NK cells, in culture.
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.
Related Flow Cytometry Resources
Demystifying Multi-Parameter Flow Cytometry
Demystifying Multi-Parameter Flow Cytometry
In this first part of a two-part webinar series, learn about flow cytometry basics including spectral overlap and compensation, how to choose a fluorochrome, and basic data analysis.
Turning Flow Cytometry Upside Down and Inside Out
Turning Flow Cytometry Upside Down and Inside Out
In this second part of a two-part webinar series, learn how to apply the basic concepts of flow cytometry to topics such as cell proliferation, dead cell exclusion, and intracellular staining.
Cell Markers Guide for Human Immune Cell Characterization
Cell Markers Guide for Human Immune Cell Characterization
This miniposter outlines the main phenotyping markers cited in the scientific literature that are used to distinguish different immune cell types or specific immune cell subsets.
CD4+ T Cell Subsets Brochure
CD4+ T Cell Subsets Brochure
This brochure describes our workflow solutions for the isolation, differentiation, and identification of the different CD4+ T cell subsets.
B Cells Brochure
B Cells Brochure
This brochure provides a detailed graphic outlining B cell development and highlights our selection of fluorochrome-conjugated antibodies for detecting and characterizing human and mouse B cell subsets.
Custom Antibody Services
Custom Antibody Services
Utilize our antibody development experts for your custom antibody needs. Our antibody services include immunization and purification, recombinant conversion, and engineering services.
Background Information
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.
1) 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.
2) 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.
3) 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.
4) 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.
5) 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.
6) 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.
