Abstract
In Procedure A 4, page 3 of our Bio-protocol paper https://bio-protocol.org/e2531, the dilute concentration “100 μg/μl” is incorrect, should be “10 μg/μl”. Consequently, the volume added into DCs is “40 μl”, the final volume should be “1.04 ml”. Figure 1 has the same mistake. The correct text should be: 4. On Day 6, the cells are pulsed with OVA (400 μg/ml) for 24 h. In detail, OVA is diluted by PBS at 10 μg/μl first and then added into DCs at 40 μl per well. The final volume is 1.04 ml per well. At least 2 wells of cells are untreated with OVA as negative control for the reaction. The revised Figure 1 is: Figure 1. OVA pulsed DCs preparation. Bone marrow cells are isolated from B6 mice and cultured in completed medium with GM-CSF and IL-4 for 7 days. Half/half medium is changed on Day 2. BM-DCs are stimulated with IFN-γ on Day 5 for 48 h and are pulsed with OVA on Day 6 for 24 h. Cells are harvested on Day 7 and washed thoroughly before using.
References
If you have any questions/comments about this protocol, you are highly recommended to post here. We will invite the authors of this protocol as well as some of its users to address your questions/comments. To make it easier for them to help you, you are encouraged to post your data including images for the troubleshooting.