Multiparameter Immunophenotyping of Influenza-Infected Ferret Lymph Nodes

Lymph node cell suspensions from influenza infected ferret were stained with the following panel: live/dead Blue (Thermo Fisher), CD79a PerCP-Cy5.5 (HM47; BioLegend)^{24 (link)}, CD8 AF700 (OKT8; Thermo)^{24 (link)}, CD4 FITC (from CSIRO)^{25 (link)}, BCL6 AF647 (K112-91; BD), BCL6 AF647 (IG191E/A8; BioLegend), CXCR5 BV421 (L138D7; BioLegend), CXCR5 BB515 (RF8B2; BD), CXCR5 PE (2G8, BD); CXCR5 Biotin (in-house), Streptavidin BV421 (BD), PD-1 BV786 (29F.1A12; BioLegend), PD-1 BV421 (EH12.2H7; BioLegend), PD-1 PE (in-house). For BCL6 staining, cells were fixed, permeabilized, and stained using the BD Transcription Factor Buffer kit (BD) according to the manufacturer’s instructions. Macaque LN suspensions were stained with Live/dead Aqua (Thermo Fisher), CD4 BV605 (L200; BD), CXCR5 PECy7 (MU5UBEE, Thermo Fisher), and CD3 BUV737 (SP34-2, BD). All samples were acquired on a BD LSR Fortessa using BD FACS Diva and data was analyzed in FlowJo v10.

Free full text: Click here

Jiang W., Wong J., Tan H.X., Kelly H.G., Whitney P.G., Barr I., Layton D.S., Kent S.J., Wheatley A.K, & Juno J.A. (2021). Screening and development of monoclonal antibodies for identification of ferret T follicular helper cells. Scientific Reports, 11, 1864.

Publication 2021

live/dead Blue BCL6 AF647 CXCR5 BV421 CXCR5 PE CXCR5 Biotin Streptavidin BV421 PD-1 BV786 PD-1 BV421 Live/dead Aqua CD4 BV605 CD3 BUV737 BD LSR Fortessa BD FACS Diva

Af647 Bcl6 Biotin Buffer Cd79a Cells Cxcr5 Cy5 5 Ferret Fitc Influenza Lymph node Macaque Streptavidin Transcription factor

Corresponding Organization :

Other organizations : Peter Doherty Institute, University of Melbourne, CSIRO Health and Biosecurity, Australian Centre for Disease Preparedness

Top 5 similar protocols

Variable analysis

independent variables

Influenza infection in ferrets

dependent variables

Immune cell populations in lymph node cell suspensions, as measured by flow cytometry using the following markers: live/dead, CD79a, CD8, CD4, BCL6, CXCR5, PD-1

control variables

Macaque lymph node cell suspensions stained with live/dead, CD4, CXCR5, and CD3 markers

controls

Positive control: Macaque lymph node cell suspensions
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!