Shop All Fixation & Permeabilization Reagents

FIX & PERM™ Cell Permeabilization Kit (Invitrogen™)

The FIX & PERM® Cell Permeabilization Kit is used in flow cytometry for fixing and permeabilizing cells in suspension, allowing access of antibody to intracellular targets without sacrificing scatter characteristics.

The FIX & PERM® Cell Permeabilization Kit is:

• Flexible—compatible with multiple sample types and cellular antigens
• Robust—scatter pattern of cells unaffected
• Efficient—simultaneous detection of intracellular and surface markers

View a selection guide for all available FIX & PERM® products and products related to sample preparation (including cell lysis and preservation solutions, blocking reagents, and information about the use of Dynabeads® cell separation products for cell isolation).

The FIX & PERM® formula was specifically designed to reduce background staining and allow simultaneous addition of fluorochrome-labeled antibodies during the permeabilization step.The FIX & PERM® Cell Permeabilization Kit is suitable for various biological samples and has been validated for use with cell lines, whole blood, and bone marrow. The FIX & PERM® kit has been shown to accurately identify previously undetectable intracellular markers. Unlike other fixation and permeabilization reagents available on the market, the FIX & PERM® staining protocol leaves the morphological scatter characteristics of the cells intact.

How It Works
The FIX & PERM® Cell Permeabilization Kit contains 2 reagents: Fixation Medium (Medium A) and Permeabilization Medium (Medium B). Cells are incubated for 15 minutes at room temperature with Fixation Medium, washed, and then incubated with the Permeabilization Medium and fluorochrome-labeled antibody. Fixation Medium and Permeabilization Medium are also available separately (Cat. Nos. GAS001S100 & GAS002S100).

eBioscience™ Foxp3 / Transcription Factor Staining Buffer Set (Invitrogen™)

The Foxp3 Transcription Factor Staining Buffer Set has been formulated and optimized for staining with antibodies to transcription factors and nuclear proteins, such as Foxp3 and Ki-67, as well as cytokines and chemokines.

For your convenience, we have provided these protocols for preparing your samples for intracellular antigen detection on a flow cytometer.

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis

Fixation Medium (Medium A) (Invitrogen™)

Fixation Medium (Medium A) is a component of the FIX & PERM® Cell Permeabilization Kit. The kit is used in flow cytometry for fixing and permeabilizing cells in suspension, allowing access of antibody to intracellular targets without sacrificing scatter characteristics.

The FIX & PERM® Cell Permeabilization Kit is:

• Flexible—compatible with multiple sample types and cellular antigens
• Robust—scatter pattern of cells unaffected
• Efficient—simultaneous detection of intracellular and surface markers

View a selection guide for all available FIX & PERM® products and products related to sample preparation (including cell lysis and preservation solutions, blocking reagents, and information about the use of Dynabeads® cell separation products for cell isolation).

The Fixation Medium formula was specifically designed to reduce background staining and allow simultaneous addition of fluorochrome-labeled antibodies during the permeabilization step.

How It Works
Cells are incubated for 15 minutes at room temperature with the Fixation Medium and washed. Cells can then be incubated with Permeabilization Medium (Medium B) (Cat. No. GAS002S100) and fluorochrome-labeled antibody. Both Fixation Medium and Permeabilization Medium are included in the FIX & PERM® Cell Permeabilization Kit.

eBioscience™ IC Fixation Buffer (Invitrogen™)

The Intracellular (IC) Fixation Buffer can be used to fix cells prior to permeabilization with the Permeabilization Buffer (cat. 00-8333), when performing intracellular staining of antigens located in the cytoplasm and secretory pathway, such as cytokines, chemokines and growth factors. This buffer, when used as instructed, is compatible with surface staining fluorochrome-conjugated antibodies and can be used as a short-term storage buffer for fluorochrome (including tandem dyes)-stained cells for up to 3 days at 2-8°C (stored in the dark).

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis

Image-iT™ Fixative Solution (4% formaldehyde, methanol-free) (Invitrogen™)

Image-iT™ Fixative Solution is a ready-to-use neutral PBS solution containing 4% formaldehyde (w/v) that is free of methanol. Its unique formulation makes it ideal for use with fluorescent proteins, GFP, RFP, and other small dye fluorophores. No more trying to mix toxic paraformaldehyde powder or dilute a concentrated solution with non-optimal pH. Just add to cells/tissue and incubate for 10-15 minutes at room temperature. Ideal for use with any antibody-based detection method, including immunocytochemistry (ICC), immunofluorescence (IF), and immunohistochemistry (IHC). Also available as part of the complete Image-iT™ Fixation/Permeabilization Kit.

• Ready-to-use, with room temperature storage
• High-purity 4% formaldehyde in PBS at neutral pH
• Methanol-free
• Optimized for secondary antibody labeling, fixed-cell dye staining, and GFP applications

See other ready-to-use, reagents for cell staining
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Fluoromount-G™ (Invitrogen™)

Fluoromount-G™ is a clear liquid medium designed for use in mounting slides following immunofluorescent staining. This water-soluble Fluoromount-G™ is used to mount slides in which the final step of staining is aqueous. It forms a semi-permanent seal for prolonged storage of slides at 2-8°C. Fluoromount-G™ does not fluoresce and may reduce the amount of fluorochrome quenching during fluorescence microscopy. This medium is supplied as a 1X stock solution containing sodium azide.

Reported Application
Immunohistochemical Staining, Immunocytochemistry

eBioscience™ 1-step Fix/Lyse Solution (10X) (Invitrogen™)

The eBioscience 1-step Fix/Lyse Solution enables lysis of red blood cells after staining peripheral blood cells with fluorochrome-conjugated antibodies. This solution has been specially formulated to lyse non-nucleated erythrocytes while maintaining a fixed and labeled leukocyte population. Therefore, whole blood samples can be stained for the appropriate markers, RBC-lysed and fixed, washed, and then analyzed by flow cytometry. Isolation of mononuclear cells by gradient centrifugation (e.g. Ficoll-Paque density gradient separation) is not required when using this solution. The eBioscience 1-step Fix/Lyse Solution is compatible with antibodies conjugated to organic dyes and tandem dyes.

Reported Application
Flow Cytometric Analysis

eBioscience™ Intracellular Fixation & Permeabilization Buffer Set (Invitrogen™)

The Intracellular Fixation & Permeabilization Buffer Set is designed for use in intracellular staining and flow cytometric analysis and has been specially formulated to reduce non-specific staining of fluorochrome-labelled antibodies and increase fluorescence signal to noise ratios. In the first step live cells can be “fixed” with the Fixation Buffer, which cross- links proteins. The second step, using the Permeabilization Buffer, creates holes in the membrane thereby allowing the intracellular staining antibodies to enter the cell effectively. Subsequent washing steps, antibody additions, and incubations after cell permeabilization should be performed using the Permeabilization Buffer. The final washing step and cell resuspension prior to running the sample is done using Flow Cytometry Staining Buffer (cat. 00-4222). This kit is commonly used for staining both cytoplasmic proteins as well as proteins in the secretory pathway such as chemokines and cytokines. It is not recommended for intranuclear staining. Instead, please refer to the Foxp3 Transcription Factor Buffer Set (cat. 00-5523) for nuclear staining.

Before the fixation step, it is recommended to label the live cells using one of the Fixable Viability Dyes (eFluor™ 450, 506, 660, 780, 520, or 455UV) and then proceed with fixation.

Reported Application
Flow Cytometric Analysis, Intracellular Staining Followed by Flow Cytometric Analysis

ViewRNA™ Cell Plus Fixation/Permeabilization Buffer Set (Invitrogen™)

The ViewRNA™ Cell Plus Fixation/Permeabilization Buffer Set is a component of the ViewRNA™ Cell Plus Assay kit (cat. 88-19000-99). These buffers can be used to evaluate antibody performance following the fixation and permeabilization steps of the assay. Note that additional changes to antibody performance may be observed following the hybridization steps of the ViewRNA™ Cell Plus Assay.

Please refer to the ViewRNA™ Cell Plus Assay protocol on the product data sheet for instructions for use.

eBioscience™ Fixation/Permeabilization Concentrate (Invitrogen™)

The Fixation/Permeabilization Concentrate is intended to be used with antibodies to transcription factors and nuclear proteins, such as Foxp3 and Ki-67, as well as cytokines and chemokines. This must be used in combination with the Fixation/Permeabilization Diluent (cat. 00-5223) and the Permeabilization Buffer (10X) (cat. 00-8333). Alternatively, all buffers required for staining with these antibodies can be purchased as a set in the Foxp3 Transcription Factor Staining Buffer Set (cat. 00-5523).

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis

eBioscience™ Permeabilization Buffer (10X) (Invitrogen™)

The Permeabilization Buffer (10X), in conjunction with the Intracellular (IC) Fixation Buffer (cat. 00-8222) or Foxp3 Transcription Factor Fixation/Permeabilization Concentrate and Diluent (cat. 00-5521), can be used to fix and permeabilize cells prior to performing intracellular staining of cytokines and other cytoplasmic and/or nuclear antigens. These solutions have been specially formulated for reducing non-specific staining of fluorochrome-labeled antibodies and increasing fluorescence signal to noise ratios.

In intracellular staining protocols, fixation is performed first, followed by permeabilization. Subsequent washing steps, antibody dilutions, and incubations after permeabilization should use the Permeabilization Buffer. The final washing step and cell resuspension prior to running the sample is done using Flow Cytometry Staining Buffer.

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis

Pierce™ 16% Formaldehyde (w/v), Methanol-free (Thermo Scientific™)

Thermo Scientific Pierce Formaldehyde Ampules are vials of high-quality, methanol-free formaldehyde for use as a general IHC fixative and reversible amine-crosslinking agent for protein and nucleic acid tissue samples.

The availability of a high-quality, ampule-sealed solution of formaldehyde saves valuable time and eliminates the need and hazards associated with preparing the solution directly from paraformaldehyde. Pierce Formaldehyde is carefully prepared directly from granular paraformaldehyde and sealed under an inert atmosphere in 1 mL and 10 mL ampules. Ampule packaging stabilizes the preparation by protecting the solution from both air oxidation and light, allowing access to "fresh" formaldehyde every time.

Features of Methanol-free 16% Formaldehyde Ampules:

Fresh—formaldehyde solution remains stable and free of contamination in an amber pre-scored ampule.
Sized Right—package sizes geared for applications ranging from culture-targeted cell fixing to in-cell protein-based applications.
Convenient Starting Concentration—16% w/v formaldehyde solution simplifies dilution to the required working concentration.
Minimizes Waste—1 mL and 10 mL package sizes allow selection of product that will result in less waste and minimize disposal concerns.

Application-targeted ampule packaging
The package sizes of the 16% Formaldehyde Solution (formalin) are targeted to both low and high volume users. The appropriate package size is selected based on the most common applications requiring a fresh, methanol-free formaldehyde solution. The 16% (w/v) solution simplifies the dilution for each respective application and the application-directed packaging minimizes waste.

• 1-mL ampules are intended for protein-based in cell applications such as Chromatin Immunoprecipitation (ChIP) or protein interaction discovery.
• 10-mL ampules are intended for bioimaging (high content screening/analysis) such as fixing cells on a culture plate before image analysis.

"Prepared fresh" Sealed in Ampules
Our formaldehyde solution is prepared as a 16% (w/v) solution of paraformaldehyde (pure granular solid) in distilled/deionized water. Solution stabilization is important because formaldehyde solutions oxidize to formic acid and eventually repolymerize to paraformaldehyde. Most protocols requiring methanol-free formaldehyde suggest that the formaldehyde be "prepared fresh". This formaldehyde solution is sealed in ampules immediately after formulation under an inert atmosphere. Ampule packaging stabilizes the preparation allowing access to "fresh" formaldehyde each time.

For added safety when opening glass ampules, consider using our Ampule Breakers, which are disposable safety devices that protect the fingers when breaking open a glass ampule.

Fluoromount-G™, with DAPI (Invitrogen™)

Fluoromount-G™, with DAPI is a clear liquid medium containing a fluorescent nuclear stain for use in mounting slides following immunofluorescent staining as well as staining double stranded DNA. This water-soluble Fluoromount-G™ is used to mount slides in which the final step of staining is aqueous. It forms a semi-permanent seal for prolonged storage of slides at 2-8°C. Fluoromount-G™, with DAPI has a maximal fluorescent emission at 455 nm and may reduce the amount of fluorochrome quenching during fluorescence microscopy. This medium is supplied as a 1X stock solution containing sodium azide.

Reported Application
Immunohistochemical Staining, Immunocytochemistry, Microscopy

PrimeFlow™ Fixation/Permeabilization Buffer Set (Invitrogen™)

The PrimeFlow™ Fixation/Permeabilization Buffer Set is a component of the PrimeFlow™ RNA Assay kit (cat. 88-18001-204 and 88-18009-204). These buffers can be used to evaluate intracellular staining antibody performance following the fixation and permeabilization steps of the assay. Note that additional changes to antibody performance may be observed after the hybridization steps of the PrimeFlow™ RNA assay.

Please refer to the PrimeFlow™ RNA Assay protocol on the product data sheet for instructions for use.

eBioscience™ Foxp3 / Transcription Factor Fixation/Permeabilization Concentrate and Diluent (Invitrogen™)

The Foxp3 Transcription Factor Fixation/Permeabilization Concentrate and Diluent solutions for staining with antibodies to transcription factors and nuclear proteins, such as Foxp3 and Ki-67, as well as cytokines and chemokines. It should be used in combination with the Permeabilization Buffer (10X) (cat. 00-8333). All three components can be purchased together in the Foxp3 / Transcription Factor Staining Buffer Set (cat. 00-5523).

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis