8–10 week old C57BL/6 mice were injected i.v. with LLC exosomes (10μg, in 100μl PBS) every 3 days for 2 weeks. Mice were fasted for 12 hours prior to intraperitoneal injection with 18FDG. One hour after injection, the mice were euthanized, lungs were harvested and measured on a Biodex Atomlab™ 500 for radioactivity. Lung tissue was enzymatically digested (collagenase (5g/L), Hyaluronidase (0.4g/L), DNAse I (0.15g/L)) for 20 minutes with rotation at 37°C. Following digestion, RBC’s were lysed using ACK. Cells were stained with primary Biotin anti-mouse CD19 (Biolegend,115504), secondary Streptavidin MicroBeads (Miltenyl Biotec,130-048-101), CD8a (Ly-2) MicroBeads (Miltenyl Biotec,130-049-401), CD4 (L3T4) (Miltenyl Biotec,130-049-201) for 15 minutes. Cells were then magnetically separated on an autoMACS Pro separator (Miltenyl Biotec) using the positive selection protocol. The negative fraction was then collected and read on a Biodex Atomlab™ 500 for radioactivity. Cells were then resuspended in Trizol and saved in the −80°C for RT-PCR.
Biotin anti mouse cd19
Manufactured by BioLegend
Biotin anti-mouse CD19 is a lab equipment product that specifically binds to the CD19 antigen on mouse cells. It is used in flow cytometry and other immunological applications to identify and quantify CD19-positive cells.
Automatically generated - may contain errors
Lab products found in correlation
Atomlab 500, by Mirion Technologies (1 mentions)
Cd4 l3t4, by Miltenyi Biotec (1 mentions)
Automacs pro separator, by Miltenyi Biotec (1 mentions)
Streptavidin microbeads, by Miltenyi Biotec (1 mentions)
Cd8a ly 2 microbeads, by Miltenyi Biotec (1 mentions)
Magnabind streptavidin, by Thermo Fisher Scientific (1 mentions)
Biotin anti mouse cd11c, by BioLegend (1 mentions)
Biotin anti mouse human cd11b, by BioLegend (1 mentions)
Ls column, by Miltenyi Biotec (1 mentions)
Mojosort streptavidin nanobeads, by BioLegend (1 mentions)
2 protocols using biotin anti mouse cd19
1
Quantification of Tumor-Derived Exosome Uptake
2
Isolation and Activation of Regulatory T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice at age between 7 to 10 weeks were sacrificed and the spleens were isolated. After red blood cell lysis, cells were incubated with biotin anti-mouse CD8α (100,704, BioLegend), biotin anti-mouse I-A/I-E (107,604, BioLegend), biotin anti-mouse NK1.1 (108,704, BioLegend), biotin anti-mouse/human B220 (103,204, BioLegend), biotin anti-mouse CD49b (103,522, BioLegend), biotin anti-mouse CD19 (115,504, BioLegend), biotin anti-mouse/human CD11b (101,204, BioLegend), and biotin anti-mouse CD11c (117,304, BioLegend). Antibody-bound cells were then negatively selected by using MagnaBind™ Streptavidin (21,344, Thermo Fisher Scientific). Subsequently, Biotin anti-mouse CD25 (102,004, BioLegend) and MojoSort™ streptavidin nanobeads (480,016, BioLegend) were used for positive selection. The cells were purified by magnetic separation using LS Columns (130–042-401, Miltenyi Biotec) according to the manufacturer’s instructions.
The isolated cells were activated with plate-bound anti-CD3ε (145-2C11; 5 μg/ml) and soluble anti-CD28 (37.51; 2 μg/ml). For tTreg culture, mouse recombinant IL-2 (50 ng/ml), human recombinant TGF-β1 (5 ng/ml), XMG1.2 (10 μg/ml), and 11B11 (10 μg/ml) were added to the cell culture medium. All cytokines for differentiation were purchased from eBioscience.
The isolated cells were activated with plate-bound anti-CD3ε (145-2C11; 5 μg/ml) and soluble anti-CD28 (37.51; 2 μg/ml). For tTreg culture, mouse recombinant IL-2 (50 ng/ml), human recombinant TGF-β1 (5 ng/ml), XMG1.2 (10 μg/ml), and 11B11 (10 μg/ml) were added to the cell culture medium. All cytokines for differentiation were purchased from eBioscience.
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!