We pooled lungs from 10–12 (P1) newborn ZBTB16CreGFP mice (on CD45.1 background). Single cells were stained with 7-AAD Viability staining solution (Biolegend), anti-mouse CD3ε antibody (145–2C11), anti–mouse CD4 antibody (GK1.5), anti-mouse CD8 antibody (53–5.8), anti–mouse CD11b antibody (M1/70), anti-mouse CD11c (N418), anti–mouse CD19 antibody (1D3), anti-mouse B220 (2FI), anti-mouse LY6G antibody (1A8), anti-mouse NKp46 antibody (29A1.4), anti-mouse α4β7 antibody (DATK32), anti-mouse CCR6 antibody (29–2L17), anti–mouse CD127 antibody (A7R34) and anti-mouse KLRG1 antibody (2FI) (all diluted 1:100, Biolegend). The stained cells were sorted with Sony SH800S. ZBTB16+ ILC precursors were sorted as Live CD45+Lineage (CD3ε, CD4, CD8, CD11b, CD11c, CD19, B220 and Ly6G), CD127+α4β7+NKp46−KLRG1−CCR6−CD25−PD1+GFP+ cells. Using this protocol, we isolated an enriched population of 800–1000 ZBTB16+ ILC precursors. We confirmed the purity of the sorted population by intracellular staining with anti-mouse RORγt antibody (Q31–378), anti-mouse T-bet antibody (4B10), anti-mouse PD1 antibody (J43), anti-mouse Id2 antibody (17-9475-82) and anti-mouse GATA3 antibody (16E10A23) (all diluted 1:50) (Supplementary Fig. 5 A , B ). The cells were resuspended in normal saline to final concentration of 1×103 cells/100 μl. We adoptively transferred 1×103 ZBTB16+ ILC precursors via ntratracheal route.
Anti mouse nkp46 antibody
Manufactured by BioLegend
The Anti-mouse NKp46 antibody is a reagent for the identification and characterization of natural killer (NK) cells in mouse samples. NKp46 is a key activating receptor expressed on the surface of NK cells, and this antibody can be used to detect and analyze NK cell populations in flow cytometry and other immunological applications.
Automatically generated - may contain errors
2 protocols using anti mouse nkp46 antibody
1
Isolation and Transfer of ILC Precursors
2
Immunohistochemical Staining of NKp46 in Frozen Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue was fixed in 4% paraformaldehyde overnight and transferred to a 30% sucrose solution for an additional 24 hours. Afterward, the tissue was embedded in OCT compound and frozen at -80°C for 24 hours. Six-micrometer-thick tissue sections were prepared on glass slides and preserved at -80°C. At the time of immunohistochemical staining, each slide was immersed in cold acetone solution for 10 minutes for tissue preservation and blocked with 2.5% bovine serum albumin overnight. Staining was performed with 2 µg/mL anti-mouse NKp46 antibody (Bio-Legend Inc.) at 4°C overnight, and 100 µL of Alexa Fluor Goat Anti-Mouse 596 ReadyProbe reagent (Life Technologies) was placed at room temperature and was used for secondary antibody staining. Slides were rinsed with phosphate-buffered saline 8 times and stained with DAPI Fluoromount (SouthernBiotech). Tissue sections were imaged at ×20 magnification using a fluorescent microscope.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!