Proliferation of Rag1-/- pro-B cells

Rag1^−/− pro-B cells were purified using anti-CD19 magnetic beads (Miltenyi). Cells were cultured at 1×10⁶ cells/ml in 96-well plates in the presence of 10 ng/ml IL-7 (Peprotech). After 24, 48, and 72 hours, proliferation was measured by flow cytometry using AccuCount 5.27 μm Blank Particles (Spherotech). Q-VD-OPH (Sigma Aldrich) was used at 100 μM. For transduction, Rag1^−/− pro-B cells were cultured for 24 hours prior to transduction with MLP or MLP-shp53 retroviral vectors, as previously described (19 (link), 25 (link)).

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Fahl S.P., Harris B., Coffey F, & Wiest D.L. (2015). Rpl22 loss impairs the development of B lymphocytes by activating a p53-dependent checkpoint. Journal of immunology (Baltimore, Md. : 1950), 194(1), 200-209.

Publication 2014

anti-CD19 magnetic beads IL-7 AccuCount 5.27 μm Blank Particles Q-VD-OPH

Cells Flow cytometry Il 10 Pro b cells Q vd oph Rag1 Retroviral Vectors

Corresponding Organization :

Other organizations : Fox Chase Cancer Center

Top 5 similar protocols

Variable analysis

independent variables

Treatment with IL-7 (10 ng/ml)
Transduction with MLP or MLP-shp53 retroviral vectors

dependent variables

Proliferation of Rag1-/- pro-B cells, measured by flow cytometry using AccuCount 5.27 μm Blank Particles (Spherotech)

control variables

Culturing Rag1-/- pro-B cells at 1×10^6 cells/ml in 96-well plates
Incubation time points (24, 48, and 72 hours)

controls

Positive control: Not explicitly mentioned
Negative control: Q-VD-OPH (Sigma Aldrich) at 100 μM

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!