Flow cytometry and tetramer were performed as previously described55 (link). Briefly, single cell suspended splenocytes were incubated with gp33-tetramer and np396-tetramer for 15 minutes and gp-66 tetramer for 30 minutes at 37 °C. After the incubation, surface antibodies were added, including anti-CD8 or anti-CD4 along with anti-IL7R, anti-PD-1, anti-CD62L, anti-CD44, anti-KLRG-1, anti-TIM3, anti-Lag-3, and anti-2B4 antibodies (eBioscience) for 30 minutes at 4 °C. For intracellular cytokine stain single suspended splenocytes were incubated with the LCMV specific peptides gp33 and np396 or the MHC-II epitope gp61. After 1 h, Brefeldin A (eBioscience) was added, followed by an additional 5 h incubation at 37 °C. After surface stain with anti-CD8 or anti-CD4 antibodies (eBioscience) cells were fixed with 2% formalin and permeabilized with 0.1% Saponin and stained with anti-IFNγ and anti-TNF antibodies (eBioscience) for 30 min at 4 °C.
Anti il7r
Manufactured by Thermo Fisher Scientific
Anti-IL7R is a laboratory research tool used to detect and quantify the presence of the Interleukin-7 Receptor (IL7R) protein in biological samples. It functions as a specific binding agent for the IL7R, allowing for its identification and measurement.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd8, by Thermo Fisher Scientific (1 mentions)
Anti cd62l, by Thermo Fisher Scientific (1 mentions)
Anti cd44, by Thermo Fisher Scientific (1 mentions)
Brefeldin a, by Thermo Fisher Scientific (1 mentions)
Anti klrg 1, by Thermo Fisher Scientific (1 mentions)
Anti tim3, by Thermo Fisher Scientific (1 mentions)
Anti lag 3, by Thermo Fisher Scientific (1 mentions)
Anti 2b4, by Thermo Fisher Scientific (1 mentions)
Anti pd 1, by Thermo Fisher Scientific (1 mentions)
Anti cd4, by Thermo Fisher Scientific (1 mentions)
Dnase 1, by Merck Group (1 mentions)
Anti cd4, by BD (1 mentions)
2 protocols using anti il7r
1
Characterization of Antigen-Specific T Cell Responses
2
Immunostaining Protocol for Human Antibodies
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Example 1
Human Antibodies:
PE-conjugated anti-CD127, APC-conjugated anti-CD25, PerCP-conjugated anti-CD4 used for staining and in sorting was provided by Becton-Dickenson (BD Pharmingen, San Diego, Calif.). Alexa488-conjugated anti-FoxP3 was purchased from BioLegend (San Diego, Calif.) and intracellular staining was performed according to manufacturer's instructions as modified as follows: 5×105 cells were stained with cell surface markers for 30 min at 4° C. and fixed for 30 min using 1× Fix/Perm buffer. After 3 washes cells were permeabilized in Perm buffer with DNase I (Sigma-Aldrich, St. Louis, Mo.) for 30 min followed by 3 washes. Then cells were blocked with human IgG and stained with anti-human FoxP3-Alexa488 conjugated (BioLegend, San Diego, Calif. clone 206D). The following anti-mouse antibodies were purchased from the indicated sources: anti-CD4, anti-CD25 and mouse IgGi (isotype control) (BD Pharmingen, San Diego, Calif.); and anti-IL-7R (eBioscience, San Diego, Calif.).
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