Anti mouse human cd44 clone im7

Manufactured by BioLegend

Anti-mouse/human CD44 (clone IM7) is a monoclonal antibody that specifically binds to the CD44 antigen expressed on various cell types. CD44 is a cell surface glycoprotein involved in cell-cell and cell-matrix interactions.

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4 protocols using anti mouse human cd44 clone im7

Multiparametric Flow Cytometry of CD8+ T Cells

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FC receptors were blocked with anti-mouse CD16/32 (clone 93, Biolegend) before staining. Dead cells were identified using the Zombie Aqua Fixable Viability Kit (Biolegend). Cells were stained with the following antibody clones: anti-mouse CD8a (clone 53-6.7, Biolegend), anti-mouse/human CD44 (clone IM7, Biolegend), anti-mouse CD62L (clone MEL-14, eBioscience), anti-mouse/human Granzyme B (clone GB11, Biolegend), anti-mouse CTLA4 (clone UC1-4B9, Biolegend), anti-mouse CD25 (clone PC61.5, eBioscience), and anti-mouse IFN-γ (clone XMG1.2, Biolegend). Intracellular staining of Granzyme B, CTLA-4, and IFN-γ was performed using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience). To count cells, 123count eBeads (eBioscience) were added to flow cytometry tubes immediately before flow cytometer acquisition. Data was acquired on a BD LSRFortessa and analyzed in FlowJo. Cells were gated for size, single cells, living cells, and CD8⁺ cells before examination of proliferation curves (Supplementary Fig. 8c). Cells were further gated on proliferation dye⁺ cells to exclude any CD8⁺ T cells in the APC fraction before quantification of division numbers and examination of surface and intracellular proteins. Statistical analyses of results from separate mice were performed using GraphPad Prism software.

Richard A.C., Lun A.T., Lau W.W., Göttgens B., Marioni J.C, & Griffiths G.M. (2018). T cell cytolytic capacity is independent of initial stimulation strength. Nature immunology, 19(8), 849-858.

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Multiparametric Flow Cytometry for Immune Profiling

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Vaginal mucosa cells and splenocytes were analyzed by flow cytometry using the following antibodies: mouse anti-guinea pig CD8 (clone MCA752F, Bio-Rad Laboratories, Hercules, CA), mouse anti-guinea pig CD4 (clone MCA749PE, Bio-Rad Laboratories), anti-mouse CRTAM (clone 11–5, Biolegend, San Diego, CA), hamster anti-mouse PD-1 clone J43, BD Biosciences, San Jose, CA), anti-mouse/human CD44 (clone IM7, Biolegend), anti-mouse CD69 (clone H1.2F3, BD Biosciences, San Jose, CA), anti-mouse CXCR3 (clone CXCR3-173, Biolegend) and anti-mouse CD103 (clone 2E7, Biolegend). For surface staining, mAbs against various cell markers were added to a total of 1X10⁶ cells in phosphate-buffered saline containing 1% FBS and 0.1% sodium azide (fluorescence-activated cell sorter [FACS] buffer) and left for 45 minutes at 4°C. At the end of the incubation period, the cells were washed twice with FACS buffer. A total of 100,000 events were acquired by the LSRII (Becton Dickinson, Mountain View, CA), followed by analysis using FlowJo software (TreeStar, Ashland, OR).

Quadiri A., Prakash S., Dhanushkodi N.R., Singer M., Zayou L., Shaik A.M., Sun M., Suzer B., Lau L., Chilukurri A., Vahed H., Schaefer H, & BenMohamed L. (2023). Therapeutic Prime/Pull Vaccination of HSV-2 Infected Guinea Pigs with the Ribonucleotide Reductase 2 (RR2) Protein and CXCL11 Chemokine Boosts Antiviral Local Tissue-Resident and Effector Memory CD4+ and CD8+ T Cells and Protects Against Recurrent Genital Herpes. bioRxiv.

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Comprehensive Guinea Pig Immune Cell Analysis

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Vaginal mucosa cells and splenocytes were analyzed by flow cytometry using the following antibodies: mouse anti-guinea pig CD8 (clone MCA752F, Bio-Rad Laboratories, Hercules, CA), mouse anti-guinea pig CD4 (clone MCA749PE, Bio-Rad Laboratories), anti-mouse CRTAM (clone 11-5, Biolegend, San Diego, CA), hamster antimouse PD-1 clone J43, BD Biosciences, San Jose, CA), anti-mouse/human CD44 (clone IM7, Biolegend), antimouse CD69 (clone H1.2F3, BD Biosciences, San Jose, CA), anti-mouse CXCR3 (clone CXCR3-173, Biolegend) and anti-mouse CD103 (clone 2E7, Biolegend). For surface staining, mAbs against various cell markers were added to a total of 1X10 6 cells in phosphate-buffered saline containing 1% FBS and 0.1% sodium azide (fluorescence-activated cell sorter [FACS] buffer) and left for 45 minutes at 4°C. At the end of the incubation period, the cells were washed twice with FACS buffer. A total of 100,000 events were acquired by the LSRII (Becton Dickinson, Mountain View, CA), followed by analysis using FlowJo software (TreeStar, Ashland, OR).

Quadiri A., Prakash S., Dhanushkodi N.R., Singer M., Zayou L., Shaik A.M., Sun M., Suzer B., Lau L.S.L., Chilukurri A., Vahed H., Schaefer H, & BenMohamed L. (2024). Therapeutic prime/pull vaccination of HSV-2-infected guinea pigs with the ribonucleotide reductase 2 (RR2) protein and CXCL11 chemokine boosts antiviral local tissue-resident and effector memory CD4⁺ and CD8⁺ T cells and protects against recurrent genital herpes. Journal of virology, 98(5).

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Multiparametric Flow Cytometry of CD8+ T Cells

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Richard A.C., Lun A.T., Lau W.W., Göttgens B., Marioni J.C, & Griffiths G.M. (2018). T cell cytolytic capacity is independent of initial stimulation strength. Nature immunology, 19(8), 849-858.

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