Cytofix and Cytoperm Method

Intracellular cytokine staining with flexibility of experimental design and the confidence of established reagents

The BD Cytofix/Cytoperm™ method and related reagents address the wide-ranging experimental needs of scientists conducting basic research. With reagents for a broad selection of cytokines and multiple species, this method offers maximum flexibility in intracellular cytokine staining.

Features

  • Broad compatibility for multiple species and sample types
  • Large selection of both cytokine antibodies and fluorochromes
  • Maximum flexibility in protocol and panel design

Large selection of reagents for flexible experimental design

BD Pharmingen™ reagents for the BD Cytofix/Cytoperm™ method are optimized for intracellular cytokine staining. Antibody reagents are available for multiple species, so the protocol is suitable for analysis of human cells, and also for non-human primate, mouse, and rat samples.

Allowing individual choice and combination of cytokine and surface marker antibodies, this method can be adapted to address a particular experimental question. The wide selection of direct fluorochrome conjugates offers additional flexibility in designing multicolor experiments.

Simplified protocol

BD Cytofix/Cytoperm fixation and permeabilization reagents enable one-step fixation and permeabilization to simplify the preparation of cells for intracellular cytokine staining. Besides convenience, they offer standardization using quality control tested fixation reagents to ensure consistent results from lot to lot. BD Cytofix/Cytoperm™ Plus kits have a protein transport inhibitor for inclusion in cell culture during cell activation. Support reagents are also available, including positive control cells and isotype controls specifically tested for this method.

To make getting started even easier, we offer two starter kits based on the BD Cytofix/Cytoperm method: one for mouse and one for human cells. The starter kits provide the buffers, antibodies, controls, and protocols required to successfully activate and stain mouse or human lymphocytes for IL-2, IFN-γ, and TNF.

Kits for CD4 + T-cell phenotyping

BD Pharmingen CD4 + T-cell phenotyping kits provide a simple solution for measurement of multiple cytokines or transcription factors from a single sample using multicolor flow cytometry. Based on the BD Cytofix/Cytoperm method, these kits stain surface CD4 along with intracellular factors such as IFN-γ, IL-4, IL-17A, and FoxP3. Depending on the kit, researchers can simultaneously look at Th1, Th2, and Th17 cells; Th1 and Th17 cells; or Th17 and regulatory T cells in mouse or human model systems.

Th17 culture analyzed using Human Th1/Th2/Th17 Phenotyping Kit

Intracellular cytokine analysis in polarized Th cell subsets

Intracellular IL-17A and IL-17F staining

Intracellular cytokines in Th17 cells (time course)


Alexa Fluor® is a registered trademark of Life Technologies Corporation.

This site uses cookies. If you click accept cookies then all cookies will be written. Please review our cookies policy and configure your cookies for your experience.