Single-cell suspensions were prepared from lymphoid tissues of mice using a 70-μm cell strainer (BD Biosciences). Conjunctival tissues were first digested in RPMI (Invitrogen) with 2mg/ml DNase and 2mg/ml Collagenase (Roche) at 37°C. The following Abs were used for flow cytometry analysis: FITC-anti-CD4 (clone RM4–5), PerCP-Cy5.5-anti-CD44 (clone IM7), PE-anti-IL-7Rα (clone A7R34), APC-anti-IL-15Rα (clone 6B4C88), PE-anti-CD62L (clone MEL-14), Brilliant Violet 421-Annexin V (all above from BioLegend), PE-Cy7-anti-IL-17, and PE-anti-IL-17 (clone eBio17B7, eBioscience). For intracellular IL-17 staining, cells were stimulated with phorbol 12-myristate 13-acetate and ionomycin (Sigma-Aldrich) for 6 hours in the presence of GolgiStop™ (BD Biosciences). Stained cell were examined with an LSR II flow cytometer (BD Biosciences), and the results were analyzed using FlowJo V10 software (Tree Star).
Percp cy5.5 anti cd44 clone im7
Manufactured by BioLegend
PerCP-Cy5.5-anti-CD44 (clone IM7) is a fluorochrome-conjugated antibody that binds to the CD44 cell surface antigen. CD44 is a glycoprotein involved in cell-cell and cell-matrix interactions. The PerCP-Cy5.5 fluorochrome is used for flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Brilliant violet 421 annexin 5, by BioLegend (2 mentions)
Fitc anti cd4 clone rm4 5, by BioLegend (2 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (2 mentions)
Ionomycin, by Merck Group (2 mentions)
Phorbol 12 myristate 13 acetate, by Merck Group (2 mentions)
Collagenase, by Roche (2 mentions)
Dnase, by Roche (2 mentions)
Golgistop, by BD (2 mentions)
Lsrii flow cytometer, by BD (2 mentions)
70 μm cell strainer, by BD (2 mentions)
2 protocols using percp cy5.5 anti cd44 clone im7
1
Isolation and Analysis of Murine Lymphoid Cells
2
Isolation and Analysis of Murine Lymphoid Cells
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Single-cell suspensions were prepared from lymphoid tissues of mice using a 70-μm cell strainer (BD Biosciences). Conjunctival tissues were first digested in RPMI (Invitrogen) with 2mg/ml DNase and 2mg/ml Collagenase (Roche) at 37°C. The following Abs were used for flow cytometry analysis: FITC-anti-CD4 (clone RM4–5), PerCP-Cy5.5-anti-CD44 (clone IM7), PE-anti-IL-7Rα (clone A7R34), APC-anti-IL-15Rα (clone 6B4C88), PE-anti-CD62L (clone MEL-14), Brilliant Violet 421-Annexin V (all above from BioLegend), PE-Cy7-anti-IL-17, and PE-anti-IL-17 (clone eBio17B7, eBioscience). For intracellular IL-17 staining, cells were stimulated with phorbol 12-myristate 13-acetate and ionomycin (Sigma-Aldrich) for 6 hours in the presence of GolgiStop™ (BD Biosciences). Stained cell were examined with an LSR II flow cytometer (BD Biosciences), and the results were analyzed using FlowJo V10 software (Tree Star).
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