eBioscience™ Intracellular Fixation & Permeabilization Buffer Set
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Invitrogen™

eBioscience™ Intracellular Fixation & Permeabilization Buffer Set

The Intracellular Fixation & Permeabilization Buffer Set is designed for use in intracellular staining and flow cytometric analysis and hasRead more
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Catalog number 88-8824-00
Price (USD)
226.00
Each
Add to cart
Price (USD)
226.00
Each
Add to cart
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

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Product LineeBioscience™
Product TypeFixation/Permeabilization Buffer Set
Quantity1 kit
Solution TypeFixation/Permeabilization Buffer
Unit SizeEach

Frequently asked questions (FAQs)

Do you offer an alternative for the discontinued 3-Germ Layer ICC Kit (Cat. No. A25538)?

We do not have a direct alternative for the discontinued 3-Germ Layer ICC Kit (Cat. No. A25538). However, we do have alternative primary and secondary antibodies, as well as reagents, that can be used for trilineage differentiation. Please note that we have not internally validated the use of all these reagents together.

We recommend the following primary antibodies:

Alpha-Smooth Muscle Actin Monoclonal Antibody (1A4 (asm-1)) (Cat. No. MA5-11547)

alpha-Fetoprotein Monoclonal Antibody (AFP3) (Cat. No. 14-6583-80)

beta-3 Tubulin Monoclonal Antibody (2G10) (Cat. No. MA1-118)

We recommend the following secondary antibodies:

Goat anti-Mouse IgG2a Cross-Adsorbed, Alexa Fluor 555 (Cat. No. A21137)

Goat anti-Mouse IgG2a Cross-Adsorbed, Alexa Fluor 594 (Cat. No. A21135)

https://www.thermofisher.com/antibody/product/Goat-anti-Mouse-IgG1-Cross-Adsorbed-Secondary-Antibody-Polyclonal/A-21121

We recommend using the following reagents:

https://www.thermofisher.com/order/catalog/product/88-8824-00

NucBlue Fixed Cell ReadyProbes Reagent (DAPI) (Cat. No. R37606)

Blocker BSA (Cat. No. 37520)

DPBS (10X), no calcium, no magnesium (Cat. No. 14200075)

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

What fixative should I use to fix my cells or tissues for antibody labeling?

Aldehyde-based fixatives (e.g., formaldehyde, glutaraldehyde) crosslink various cellular components, which helps retain proteins and cell morphology, but some antigens can be masked by the crosslinking, requiring antigen retrieval methods to unmask the antigen binding sites. Also, aldehyde-based fixation requires permeabilization to allow entry of antibodies. Organic solvents such as methanol and acetone condense proteins and permeabilize the cells, but cell morphology can be compromised. Surface antigens can be labeled without fixation.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can eVolve-conjugated antibodies be used in combination with intracellular staining?

Yes! eVolve conjugates were designed to be compatible with eBioscience's intracellular staining buffers including the Foxp3/Transcription Factor Staining Buffer Set (Cat. No. 00-5523-00) and the Intracellular Fixation and Permeabilization Buffer Set (Cat. No. 88-8824-00).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.