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eBioscience™ Intracellular Fixation & Permeabilization Buffer Set - FAQs

查看更多产品信息 eBioscience™ Intracellular Fixation & Permeabilization Buffer Set - FAQs (88-8824-00)

4 个常见问题解答

我应该使用哪种固定剂来固定自己的细胞或组织，从而进行抗体标记？

醛类固定剂（例如甲醛、戊二醛）可与各种细胞成分交联，这有助于保持蛋白质和细胞的形态。但一些抗原可能会因交联而被掩藏，需要通过抗原修复的方式暴露掩藏的抗原结合位点。此外，醛类固定剂需要通过通透处理使抗体进入细胞。甲醇和丙酮等有机溶剂可凝聚蛋白质并通透细胞，但细胞形态会受到破坏。表面抗原无需固定即可标记。

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.