The following antibodies were prepared for flow cytometry: PerCP-Cy5.5–conjugated anti-mouse CD4, PE-conjugated anti-mouse IL-17A, FITC-conjugated anti-mouse IFN-γ, PE-conjugated anti-mouse CD25 (BD Bioscience), and Alexa-647–conjugated anti-mouse Foxp3 (Biolegend). Spinal cord cells were stained by anti-CD4 and F4/80 antibodies (Biolegend). CD4+ T cells were cultured with 100 nM PMA (Sigma), 1 µM inonomycin (Sigma), and GolgiPlug (BD Bioscience) for 2 h at 37°C. Intracellular staining (IFN-γ, IL-17A, and Foxp3) was performed after permeation and fixation of cells by using either the Cytofix/Cytoperm intracellular staining kit (BD Bioscience; for IFN-γ and IL-17A) or the Foxp3/Transcription factor staining buffer set (Affymetrix; for Foxp3).
Pe conjugated anti mouse cd25
Manufactured by BD
Sourced in United States
PE-conjugated anti-mouse CD25 is a laboratory reagent used to detect the expression of CD25, also known as the interleukin-2 receptor alpha chain, on mouse cells. This product is conjugated with the fluorescent dye phycoerythrin (PE) to enable flow cytometric analysis of CD25-positive cells.
Automatically generated - may contain errors
Lab products found in correlation
Golgiplug, by BD (2 mentions)
Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (2 mentions)
Percp cy5.5 conjugated anti mouse cd4, by BD (1 mentions)
Pe conjugated anti mouse il 17a, by BD (1 mentions)
Cytofix cytoperm intracellular staining kit, by BD (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Percp conjugated anti mouse cd4, by BD (1 mentions)
Fitc conjugated anti mouse cd44, by BD (1 mentions)
Apc conjugated anti mouse cd4, by BD (1 mentions)
Facslyric, by BD (1 mentions)
4 protocols using pe conjugated anti mouse cd25
1
Multiparametric Flow Cytometry Analysis of Immune Cells
2
Isolation of Naïve CD4+ T Cells for Tregs Induction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The same mice were used for isolation of naïve CD4+ T cells, which were purified from isolated autologous MNCs from EAE model mice using a CD4+ T cell magnetic bead separation kit (Catalog Number: 130-104-453, Miltenyi Biotec). The purity of the naïve CD4+ T cells was determined using peridinin chlorophyll protein (PerCP)-conjugated anti-mouse CD4 (Catalog Number: 550954), PE-conjugated anti-mouse CD25 (Catalog Number: 101904), and APC-conjugated anti-mouse CD127 MoAb (Catalog Number: 564175, BD Biosciences, San Jose, CA, USA) antibodies and a flow sorter, and only cells with purity > 90% were used for further experimentation; then, the naïve CD4+ T cells were further cocultured with the sCD40L-activated B cells to induce CD4+CD25+ Tregs.
3
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.
4
Flow Cytometric Analysis of T Cell Subsets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
T cells were treated with GolgiPlug (2 μg/mL, BD Biosciences, Franklin Lakes, NJ, USA), 4 h prior to the end of the 48 h culture, to block the secretion of cytokines. Then, the T cells were collected and resuspended in fluorescence activating cell sorting (FACS) staining buffer (0.09% sodium azide, 1% FBS, 1 × PBS base) at 5 × 105 cells/mL. A total of 5 × 105 cells (per sample) were stained with APC-conjugated anti-mouse CD4 (BD Biosciences) and PE-conjugated anti-mouse CD25 (BD Biosciences) Abs for 30 min at 4 °C. For intracellular staining, T cells were fixed and permeabilized, using Foxp3/Transcription Factor Staining Buffer Set (Invitrogen, Carlsbad, CA, USA), and then stained with Alexa Fluor 488-conjugated anti-mouse Foxp3 (BD Biosciences) and PerCP-CyTM 5.5 anti-mouse IL-17A (BD Biosciences) Abs, for 30 min, at 4°C. After staining, the cells were washed and resuspended in a FACS-staining buffer with 4% formaldehyde, and then analyzed, using FACSLyric (BD Biosciences) and FlowJo software version 10 (BD Biosciences). Gating strategy of CD4+IL-17+ T cells and CD4+CD25+Foxp3+ T cells are provided in Supplementary Figure S1 .
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!