Image-iT™ 4% Formaldehyde Fixative Solution in PBS (methanol-free), 20 mL - FAQs

查看更多产品信息 Image-iT™ Fixative Solutions - FAQs (R37814, I28700, I28800, I28900, FB002)

5 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。

The Image-iT Fixation/Permeabilization Kit includes Triton X-100 as the permeabilizing reagent. May I substitute other reagents for permeabilizing?

Yes. Aldehyde-based fixation is compatible with other permeabilizing reagents, such as saponin.

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

Is the Image-iT Fixative Solution alcohol-free?

Yes, it is alcohol-free.

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

Can I add the Image-iT Fixative Solution in with the media?

We do not recommend doing this. The Image-iT Fixative Solution is 4% formaldehdye in PBS, so mixing it in directly with media would dilute the final concentration of fixative and this may result in less than optimal fixation. If you need to add fixative directly to the medium, consider using a fixative at a higher concentration such as formaldehyde at 16% (Cat. No. 28906) or at 37% (Cat. No. 9311).

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.