APC-conjugated anti-mouse CD8a (53–6.7), FITC-conjugated anti-mouse TNF-α, allophycocyanin-conjugated anti-mouse IFN-γ, FITC-conjugated anti-mouse CD49d, FITC-conjugated anti-mouse CD44 and Golgi transport inhibitor (brefeldin A) were purchased from BD Biosciences. Allophycocyanin-conjugated and PE-conjugated H-2Kb/gB498–505 (SSIEFARL) tetramers were provided by the National Institutes of Health Tetramer Core Facility (Emory University, Atlanta, GA). Recombinant mouse Gal-9 was provided by GalPharma, Japan. CD8 T cell isolation kit was obtained from Miltenyi Biotec. Primary antibodies Rat Anti-Mouse CD8a and Rabbit Anti-Glial Fibrillary Acidic Protein (GFAP) for immunohistochmeistry staining were purchased from BD Biosceince and DAKO respectively. The secondary antibodies Donkey Anti-Rat IgG (H+L) and Donkey Anti Rabbit IgG (H+L) were purchased from Jackson Immunoresearch.
Fitc conjugated anti mouse cd44
Manufactured by BD
The FITC-conjugated anti-mouse CD44 is a monoclonal antibody that binds to the CD44 antigen expressed on the surface of mouse cells. The FITC (Fluorescein Isothiocyanate) fluorescent label allows for the detection and analysis of CD44-positive cells using flow cytometry or other fluorescence-based techniques.
Automatically generated - may contain errors
Lab products found in correlation
Donkey anti rabbit igg h l, by Jackson ImmunoResearch (1 mentions)
Cd8 t cell isolation kit, by Miltenyi Biotec (1 mentions)
Allophycocyanin conjugated anti mouse ifn γ, by BD (1 mentions)
Donkey anti rat igg h l, by Jackson ImmunoResearch (1 mentions)
Brefeldin a, by BD (1 mentions)
Pe conjugated anti mouse cd25, by BD (1 mentions)
3 protocols using fitc conjugated anti mouse cd44
1
Multiparameter Immune Cell Analysis
2
Immunofluorescence Staining of Tumor Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tumor tissue samples were prepared from frozen sections by cryostat sectioning, and 5-μm sections were stained as previously reported (30 (link)). FITC-conjugated anti-mouse CD44 and PE-conjugated anti-mouse CD25 or matching isotype controls (all from BD Biosciences, San Jose, CA) were used. Imaging was performed using a ZEISS LSM-710 confocal microscope (Zena, Germany). Images were analyzed by ImageJ software, https://imagej.net >Fiji. The co-localization index was expressed by Mander's coefficient. A value close to 1 indicates reliable co-localization.
3
CD44 Expression and Tumor Targeting Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The CD44 expression levels in HepG2 and MCF-7 cells were detected by flow cytometry. Briefly, cells were cultured in 24-well plates, and then washed with blocking buffer containing of 0.2% (v/v) FBS and 0.02% (v/v) sodium azide, and incubated with fluorescein isothiocyanate (FITC)-conjugated anti-mouse CD44 (20 mL, BD) and FITC-conjugated mouse IgG (20 mL, BD) at 4°C for 30 minutes. Subsequently, cells were treated with 200 μL of 4% paraformaldehyde, and analyzed by flow cytometry.
The CD44-mediated active tumor targeting effect was further verified by CLSM. In brief, HepG2 and MCF-7 cells were treated with C6-labeled MSNs-GOx/PLL/HA nanoparticles for 2 hours. Then, cells were fixed with 4% paraformaldehyde, and nuclei were stained with DAPI. For CD44 blocking experiments, cells were precultured with 10 mg/mL of HA in advance, and then incubated with MSNs-GOx/PLL/HA for 2 hours. Furthermore, the HA-coated nanoparticles were pretreated with HAase (1 mg/mL) for 2 hours, and then cells were incubated with the HAase-treated MSNs-GOx/PLL/HA nanoparticles for 2 hours. Cells were then treated following the same procedure as described above, and monitored using CLSM. Finally, the relative fluorescence intensity was also calculated.
The CD44-mediated active tumor targeting effect was further verified by CLSM. In brief, HepG2 and MCF-7 cells were treated with C6-labeled MSNs-GOx/PLL/HA nanoparticles for 2 hours. Then, cells were fixed with 4% paraformaldehyde, and nuclei were stained with DAPI. For CD44 blocking experiments, cells were precultured with 10 mg/mL of HA in advance, and then incubated with MSNs-GOx/PLL/HA for 2 hours. Furthermore, the HA-coated nanoparticles were pretreated with HAase (1 mg/mL) for 2 hours, and then cells were incubated with the HAase-treated MSNs-GOx/PLL/HA nanoparticles for 2 hours. Cells were then treated following the same procedure as described above, and monitored using CLSM. Finally, the relative fluorescence intensity was also calculated.
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!