Flow Cytometry Assays and Reagents

‹ Flow Cytometry Assays and Reagents

Best practices for intracellular flow cytometry

The ability to stain and detect intracellular molecules opens the door to identify distinct cell subsets as well as further characterize cell populations. Our resources, tools and products help you save time and effort and increase your efficiency when designing your flow cytometry experiments.

Target determination

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Buffer selection

When performing intracellular staining for flow cytometry the selection of buffers used for fixation and permeabilization has a significant impact on the quality and accuracy of data. We provide optimized solutions for staining cytosolic, nuclear and secreted proteins for flow cytometry.

  • Buffers for fixation and permeabilization have significant impact on the quality and accuracy of data
  • See the difference for yourself using our optimized staining buffers
Cytoplasmic staining
eBioscience Intracellular Fixation & Permeabilization Buffer Set FIX & PERM Cell Fixation & Cell Permeabilization Kit
Use
  • Stain both cytoplasmic proteins as well as proteins in the secretory pathway such as chemokines and cytokines.
  • Not recommended for intranuclear staining.
  • Use when staining intracellular markers, such as cytoplasmic or nuclear enzymes, oncoproteins, cytokines, and immunoglobulins.
  • Mild fixation and permeabilization of cells that leaves their morphological scatter characteristics intact.
    		•
    Contents
    • IC Fixation Buffer
    • Permeabilization Buffer (10X)
    • Fixation Medium (Medium A)
    • Permeabilization Medium (Medium B)
    Size 1 kit 50 tests
    Cat. No. 88-8824-00 GAS003
    Regulatory status RUO–For Research Use Only. Not for Use in Diagnostic Procedures. GPR–General Purpose Reagent.
    When to use For use in research only (i.e. research on any sample type). For use in general laboratory application, that is used to collect, prepare, and examine specimens from the human body for diagnostic purposes.
    Nuclear staining
    eBioscience Foxp3 / Transcription Factor Staining Buffer Set
    Use
    • Formulated and optimized for staining with antibodies to transcription factors and nuclear proteins, such as Foxp3 and Ki-67.
    • Also applicable to simultaneous nuclear and cytoplasmic (cytokines and transcription factors) staining.
    Contents
    • Fixation/Permeabilization Concentrate
    • Fixation/Permeabilization Diluent
    • Permeabilization Buffer (10X)
    Size 1 kit
    Cat. No. 00-5523-00
    Panel Design
    Web Tool: Flow Cytometry Panel Builder
    • Ideal for anyone designing a flow cytometry panel
    • Simplified, customizable panel design experience
    • Efficient antibody selection built on spectra visualization
    Article: Flow Cytometry Panel Design–The Basics
    • Ideal read for anyone new to designing flow cytometry panels
    • Learn the key points to consider when designing a flow cytometry panel
    • Key literature references about panel design cited
    Web reference: Optimized Flow Cytometry Multiplex Panels
    • Useful for all experience levels
    • Learn about optimized multicolor immunofluorescence panels (OMIPs)
    • Search existing published flow cytometry panels
    • Leverage a published panel to save time
    Webinar: Basics of Multicolor Flow Cytometry Panel Design
    • Ideal for beginners, 1-hour run time
    • Dr. Holden T. Maecker of Stanford University discusses the caveats of good panel design including:
    • Examples and practical applications
    • Controls and standardization
    • Relevance of panel design to mass cytometry platforms
    Biology controls

    Save time and avoid inaccurate analysis of your cell population by removing dead cells and unwanted artifacts efficiently with the Invitrogen LIVE/DEAD Fixable Dead Cell Stains.

    • Fixation and permeabilization of cells results in cell death and often leads to unwanted artifacts
    • Save time and avoid inaccurate analysis of your cell population
    • Remove these dead cells and any unwanted artifacts efficiently
    Find LIVE/DEAD kits for: UV & 405 nm lasers | 488 & 561 nm lasers | 633/635 laser
    UV & 405 nm lasers
    LIVE/DEAD Fixable Blue stain LIVE/DEAD Fixable Violet stain LIVE/DEAD Fixable Aqua stain LIVE/DEAD Fixable Yellow stain
    Basis of assay LIVE/DEAD Fixable Dead Cell Stain kits are fixable viability dyes that distinguish live cells from dead cells based on cell membrane integrity and access to available amines. Cells can then be fixed for intracellular antigen detection without loss of original cell staining pattern. Allows easy exclusion of dead cells from flow cytometry readout.
    Readout Dead cells are highly fluorescent and easy to distinguish from live cells, which are stained with minimal fluorescence
    Laser (nm) UV 405 405 405
    Ex/Em (nm) 350/450 416/451 367/526 400/575
    Multiplexable Yes Yes Yes Yes
    Dye compatibility Cannot be used in combination with Invitrogen product line such as Indo-1 (blue), DAPI, or Hoechst 33342 Cannot be used in combination with Invitrogen product line such as Pacific Blue dye, CellTrace Violet stain, FxCycle Violet stain, BV421 or eFluor 450 Cannot be used in combination with Invitrogen product line such as Pacific Green dye, F2N12S apoptosis assay, AmCyan, or BV510 Cannot be used in combination with Invitrogen product line such as Pacific Orange dye, Qdot 605, F2N12S apoptosis assay, or BV605
    Fixable Yes Yes Yes Yes
    Sample type Live cells
    Size 80 tests
    Cat. No. L34961 L34963 L34965 L34967
    Experimental controls

    Staining intracellular targets for flow cytometry requires the use of multiple antibodies that often have overlapping fluorescence emission spectra. The emission spillover must be compensated for correctly in order to ensure that the fluorescent signal being measure is due to the fluorochrome of interest and thus the cell population of interest.

    • Save time setting your experimental antibody compensation controls correctly and easily
    • Supplied in a single vial—one drop is all you need to set an antibody compensation control
    UltraComp eBeads
    Use Designed for compensation of dyes in immunophenotyping experiments using fluorescently labeled antibodies, the beads can be stained with individual fluorochrome-conjugated antibodies for use as single-color compensation controls.
    Reactivity Hamster, mouse, and rat antibodies and recognition of the kappa & lambda chains
    Format One vial, dispense as a single drop
    Laser compatibility Compatible with most standard lasers, UV to 633 nm
    Size 25 tests
    Cat. No. 01-2222-41
    Plan your experiments with the Flow Cytometry Panel Builder

    This online tool guides you through flow cytometry panel design, providing a simplified, customizable experience to fit your flow cytometry panel design needs.

    Get expert help and pricing to put together your panels

    Contact our technical sales specialists for help choosing antibodies, fluorophores, and reagents for success on the flow cytometer you’ll be using.