eBioscience™ Intracellular Fixation & Permeabilization Buffer Set

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.

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.

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.