Below is a fixation protocol for tissue culture cells for a journal article. Bri

Question

Below is a fixation protocol for tissue culture cells for a journal article. Briefly describe the role of the following reagents. PBS, paraformaldehyde. Triton X-100, and BSA. Cells were washed with PBS and fixed with 4% paraformaldehyde for 20 nun After washing with PBS, cells were permeabilized for 1 min with 0 25% Triton X-100. 0 2% BSA in PBS. Cells were incubated with primary antibodies (1:1000 dilution), including anti-alpha-actinin (Abcam ab9465) and anti-titin (M8 and T12; [31.32]) Then, secondary antibodies conjugated to either AlexaFluor 488 or 568 (Invitrogen) (1:2000 dilution) in PBS. 0.2% BSA for 1 hr each, and then three times with PBS. After the final wash, cells were mounted in ProLong Antifade with DAPI (Invitrogen). The images were acquired using a 60X oil objective (Olympus 60X/1.42) on a multi-channel spinning disc confocal microscope (1X81F-3; Olympus) and analyzed with Velocity 6 1 1 (PerkinElmer).

Explanation / Answer

Role of the following reagents in tissue culture:

Related Questions

Navigate

• Browse (All)
• Browse B
• Subjects

---

---

Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.