At various times post infection VACV infected lungs were inflated with 500 μl 50:50 OCT/PBS embedding solution and immediately snap frozen on dry ice. 7 μm thick cryo-sections of OCT embedded lung were cut by a Microtome HM 505E cryostat and prepared on super frost glass slides for immunofluorescence microscopy. Each cryo-section slide was washed with 1 ml PBS, fixed with 4 % paraformaldehyde for 15 minutes at 4°C and subsequently permeabilized with 0.1 % Triton-X100 for 5 minutes at 4°C. The sections were washed with cold PBS and incubated overnight in the dark at 4°C with rat anti-mouse CD8 APC (BD Pharmingen, clone 53-6.7, dilution 1:100) and ActinGreen (Molecular Probes, as directed by provided protocol). Sections were washed three times with PBS, mounted with cytoseal and covered with glass coverslip. The stained sections were observed and analyzed at wavelength 488 nm for FITC (green) and 647 for APC (magenta) labeling. Using an EVOS fl, (Advanced Microscopy Group inverted immunofluorescence microscope) images were captured by 20x and 40x objectives, keeping all the conditions of microscope and settings of software identical for all treatments and controls.
Rat anti mouse cd8 apc
Manufactured by BD
Sourced in United States
Rat Anti-Mouse CD8-APC is a fluorochrome-conjugated monoclonal antibody that binds to the CD8 surface antigen expressed on a subset of T lymphocytes in mice. It can be used for the identification and enumeration of CD8+ T cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Pe rat anti mouse cd4, by BD (3 mentions)
Novocyte flow cytometer, by Agilent Technologies (2 mentions)
Rat anti mouse cd3 pe, by BD (2 mentions)
Actingreen, by Thermo Fisher Scientific (1 mentions)
Mouse anti mouse nk 1.1 apc, by BD (1 mentions)
Penicillin and streptomycin, by Thermo Fisher Scientific (1 mentions)
Celltrace cfse cell proliferation kit, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Corning (1 mentions)
4 protocols using rat anti mouse cd8 apc
1
Profiling Lung CD8+ T Cells in VACV Infection
2
Cytokine Modulation in Colitis Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
KFXL was provided by Haoyisheng Pharmaceutical Company Limited (Sichuan, China). DSS was purchased from Shenzhen Lijing Biological Technology Co., Ltd (Shenzhen, China). SASP was obtained from Shanghai Xinyitianping Pharmaceutical Company, Ltd (Shanghai, China). Interleukin-1β (IL-1β), interleukin-10 (IL-10), interleukin-17 (IL-17), and epidermal growth factor (EGF) kits were from Nanjing Jiancheng Bioengineering Institute. Rat anti-mouse CD3 FITC, Rat anti-mouse CD4 PE, Rat anti-mouse CD8 APC, and Rat anti-mouse CD25 Percp-cy5.5 were acquired from BD Biosciences Co., Ltd.
3
Multicolor Flow Cytometry Immunophenotyping
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Splenocytes were stained with Mouse Anti-Mouse NK 1.1- APC (BD Biosciences, USA), Rat Anti-Mouse CD3-PE (BD Biosciences, USA), Rat Anti-Mouse CD4-PE (BD Biosciences, USA), and Rat Anti-Mouse CD8-APC (BD Biosciences) according to the manufacturer’s instructions. After staining, cells were analyzed by flow cytometry (Novocyte Flow Cytometer, ACEA Biosciences, USA). The positivity of CD8, CD4, and CD3 was determined by comparison with the defined cutoff values obtained with unstained control cells as previously described [18 (link)].
4
Evaluating T Cell Proliferation in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
T cell proliferation was assessed using a CellTrace CFSE Cell Proliferation Kit (ThermoFisher, USA). Briefly, the lymphocyte cells were isolated from spleen of C57BL/6 mouse, and then the cells were stained with CFSE (5 μM) staining solution diluted in DPBS for 20 min in a 37°C water bath. The CFSE-stained cells were added to RPMI1640 (Corning, USA) media with 10% heat-inactivated fetal bovine serum (FBS; Korea) and 100 U/ml penicillin and streptomycin (Gibco, USA) and then incubated for 5 min. Next, the cells were centrifuged at 300 ×g for 5 min, the supernatants were aspirated, and CFSE-stained cells were plated with a density of 1 × 107 cell per each well in a 6-well bottom plate and incubated for 5 days in 5% CO2 at 37°C. After 5 days, the cells were stained with Rat Anti-Mouse CD3-PE (BD Biosciences, NJ, USA), Rat Anti-Mouse CD4-PE (BD Biosciences, USA), and Rat Anti-Mouse CD8-APC (BD Biosciences). The stained cells were analyzed by flow cytometry (Novocyte Flow Cytometer, ACEA Biosciences, USA).
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!