Lungs and lung draining lymph nodes (dLN) were harvested and digested for 30 minutes at 37°C in medium containing 1mg/mL Collagenase (Type 3; MP Biomedicals, Solon, OH) and 0.02 mg/mL DNase-I (MP Biomedicals). Single cell suspensions were made and 1×106 cells/well were plated in 96-well round bottom plates. Cells were blocked with 2% rat and hamster serum for 30 minutes at 4°C. After blocking, antigen experienced CD8 T cells were identified as previously described (47 (link)). The following antibodies were used to identify antigen experienced CD8 T cell memory subsets: rat anti-mouse CD8α (53-6.7; BioLegend), rat anti-mouse CD11a (M17/4; BD Biosciences, San Jose, CA), rat anti-mouse CD103 (M290; BD Biosciences), rat anti-mouse CD69 (H1.2F3; eBioscience), rat anti-mouse CD127 (A7R34; BD Biosciences), rat anti-mouse CD62L (MEL-14; BioLegend), hamster anti-mouse KLRG1 (2F1/KLRG1; BioLegend), mouse anti-mouse CX3CR1 (SA011F11, BioLegend), and hamster anti-mouse CD27 (LG.3A10, BioLegend). Cells were fixed with BD FACS™ Lysing Solution per the manufacturer’s instructions and resuspended in PBS. Data were acquired on a LSRII (BD Biosciences) and analyzed using FlowJo software (Tree Star, Ashland, OR).
Rat anti mouse cd69 h1.2f3
Manufactured by Thermo Fisher Scientific
The Rat anti-mouse CD69 (H1.2F3) is a lab equipment product that can be used to detect the expression of the CD69 protein on the surface of mouse cells. CD69 is a cell surface receptor that is rapidly upregulated upon activation of various immune cells, including T cells, B cells, and natural killer cells. This product can be utilized in flow cytometry applications to identify and analyze CD69-expressing cells.
Automatically generated - may contain errors
Lab products found in correlation
Rat anti mouse cd8α 53 6.7, by BioLegend (2 mentions)
Rat anti mouse cd11a m17 4, by BD (2 mentions)
Rat anti mouse cd103 m290, by BD (2 mentions)
Collagenase type 3, by MP Biomedicals (1 mentions)
Sa011f11, by BioLegend (1 mentions)
Dnase 1, by MP Biomedicals (1 mentions)
Fitc conjugated peanut agglutinin pna, by Vector Laboratories (1 mentions)
Rat anti mouse cd4 gk1, by BioLegend (1 mentions)
2 protocols using rat anti mouse cd69 h1.2f3
1
Identification of CD8+ T Cell Memory Subsets
2
Multiparametric Flow Cytometry Analysis of Lung T and B Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Single-cell suspensions (1 × 106 cells) from lungs were blocked with 2% rat serum for 30 min at 4°C. Following blocking, cells were stained with the following antibodies: rat anti-mouse CD4 (GK1.5; BioLegend), rat anti-mouse CD8α (53-6.7; BioLegend), rat anti-mouse CD49d (R1-2, BioLegend), rat anti-mouse CD11a (M17/4; BD Biosciences, San Jose, CA), rat anti-mouse CD103 (M290; BD Biosciences), and rat anti-mouse CD69 (H1.2F3; eBioscience), to identify CD4 and CD8 T cell subsets. Antigen experienced T cells were identified via expression of surrogate markers as previously described (38 (link), 39 (link)). Briefly, CD11ahiCD49dpos expression was utilized to identify antigen-experienced CD4 T cells, while CD11ahiCD8αlo expression was utilized to quantify antigen-experienced CD8 T cells. To identify B cell subsets, cells were stained with rat anti-mouse CD19 (1D3; BD Biosciences), rat anti-mouse B220 (RA3-6B2; BioLegend), rat anti-mouse IgM (B7-6), and FITC-conjugated peanut agglutinin (PNA; Vector Laboratories, Burlingame, CA). Cells were then fixed with BD FACS™ Lysing Solution per manufacturer's instructions and resuspended in PBS. Data were acquired on a LSRII (BD Biosciences) and analyzed using FlowJo software (Tree Star, Ashland, OR).
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!