Invitrogen™ eBioscience™ Intracellular Fixation & Permeabilization Buffer Set

Catalog No.	Quantity
50-112-9037	1 kit

Catalog No. 50-112-9037 Supplier Invitrogen™ Supplier No. 88882400

Description

Includes

• IC Fixation Buffer
• Permeabilization Buffer (10X)

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 and allows 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
• 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 to stain cytoplasmic proteins and proteins in the secretory pathway such as chemokines and cytokines
• Not recommended for intranuclear staining. Instead, please refer to the Foxp3/Transcription Factor Buffer Set for nuclear staining
• Before the fixation step, it is recommended to label the live cells using one of the Fixable Viability Dyes and then proceed with fixation

Specifications

• Product Line: eBioscience
• Product Type: Fixn/Perm Buffer Set
• Quantity: 1 kit
• Solution Type: Fixation/Permeabilization Buffer

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)

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.

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).

For Research Use Only. Not for use in diagnostic procedures.