Isolation and Activation of Splenic B Cells

Splenic B cells were isolated from wt naïve mice in either pure B6 or non-B6 genetic background that is the same as G1XP lymphomas. B cells were purified by negative selection kit (Stem Cell Technologies, Canada), activated with anti-CD40 and IL-4 as described previously [27 (link)], and collected 4 days after culture for genomic DNA isolation which were subject to NGS. In vivo immunization and GC B cell isolation were performed as described previously [27 (link)]. The GC B cells were isolated from non-B6 genetic background that is the same as G1XP lymphomas.

Free full text: Click here

Chen Z., Gowan K., Leach S.M., Viboolsittiseri S.S., Mishra A.K., Kadoishi T., Diener K., Gao B., Jones K, & Wang J.H. (2016). Unexpected effects of different genetic backgrounds on identification of genomic rearrangements via whole-genome next generation sequencing. BMC Genomics, 17, 823.

Publication 2016

negative selection kit

B cells Cell isolation Cells Genetic background Genomic Immunization Isolation Lymphomas Mice Splenic Stem cell

Corresponding Organization :

Other organizations : University of Colorado Anschutz Medical Campus, National Jewish Health, University of Colorado Denver

Top 5 similar protocols

Variable analysis

independent variables

Genetic background (pure B6 or non-B6 genetic background that is the same as G1XP lymphomas)

dependent variables

Genomic DNA isolated from B cells

control variables

Naïve mice
B cell purification by negative selection kit
B cell activation with anti-CD40 and IL-4
B cell collection 4 days after culture
In vivo immunization and GC B cell isolation

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!