Apc anti mouse cd44

Manufactured by BioLegend

Sourced in United States

APC anti-mouse CD44 is a flow cytometry antibody that binds to the CD44 antigen expressed on the surface of mouse cells. CD44 is a cell adhesion molecule involved in cell-cell and cell-matrix interactions.

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5 protocols using apc anti mouse cd44

CD44-mediated Cellular Uptake of pCas13a

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The CD44 expression of HepG2 cells were verified with APC anti-mouse CD44 (Biolegend, USA) antibody staining and analyzed by a flow cytometer (ACEA NovoCyte, USA).
For cellular uptake analysis, HepG2 cells (1 × 10⁵ cells per well) were seeded into 12-well plates and incubated overnight. Next, pCas13a was stained with the nucleic acids probe YOYO-1. CHAIN loaded with 1 μg of pCas13a were incubated with the cells for 2 h. For the competitive assay, HepG2 cells were preincubated with free HA (10 mg/mL) and/or with galactopyranoside (1 mM) for 2 h to block the CD44 and/or ASGPR receptors. Then, cells were either analyzed by flow cytometry or washed, fixed and observed by a fluorescence microscope (Olympus, Japan).

Liu X., Yang S., Wang L., Wu X., Wang X., Ou C., Yang J., Song L., Zhou S., Wu Q, & Gong C. (2023). Hierarchically tumor-activated nanoCRISPR-Cas13a facilitates efficient microRNA disruption for multi-pathway-mediated tumor suppression. Theranostics, 13(9), 2774-2786.

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Mesenchymal Lineage Identification

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Cells were incubated with FITC anti-mouse CD90.2 (Thy1.2), APC anti-mouse CD44, Cy7 anti-mouse CD31, and PE anti-mouse CD45 (Biolegend, CA) to label mesenchymal lineage via flow cytometry analysis.

Zhuang Y., Zhao Z., Cheng M., Li M., Si J., Lin K, & Yu H. (2022). HIF-1α Regulates Osteogenesis of Periosteum-Derived Stem Cells Under Hypoxia Conditions via Modulating POSTN Expression. Frontiers in Cell and Developmental Biology, 10, 836285.

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Comprehensive Lung Cell Immunophenotyping

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Single lung-cell suspensions were incubated using a Zombie Aqua™ Fixable Viability Kit (BioLegend, San Diego, CA, USA) at RT for 30 min to discriminate dead cells for analysis. Cells were washed with DPBS containing 1% FBS, blocked with anti-CD16/CD32 (clone 2.4G2, ATCC HB-197), and then stained with FITC anti-mouse CD19 (BD Biosciences), PE anti-mouse CD69 (BioLegend), PerCP-Cy5.5 anti-mouse CD3e (BD Biosciences), PE-Cy7 anti-mouse CD103 (BioLegend), APC anti-mouse CD44 (BioLegend), APC-Cy7 anti-mouse CD8a (BioLegend), Alexa Fluor 700 anti-mouse CD4 (BD Biosciences), or BV421 anti-mouse CD45.2 (BD Biosciences) antibodies for 30 min at 4 °C. For B-cell analysis, cells were stained with FITC anti-mouse IgA (Southern Biotech), PE anti-mouse CD19 (BioLegend), PerCP-Cy5.5 anti-mouse CD38 (BioLegend), PE-Cy7 anti-mouse CD3e (BD Biosciences), biotin anti-mouse CD45.2 (eBioscience), Alexa Fluor 700 anti-mouse IgD (BioLegend), Brilliant Violet™ 421 anti-mouse CD80 (BioLegend), or streptavidin APC-Cy7 (BD Biosciences) antibodies for 30 min at 4 °C. After staining, cells were washed with DPBS containing 1% FBS and analyzed on a FACS Fortessa (BD Biosciences) using FlowJo Software Version 10 (Tree Star, Ashland, OR, USA). The gating strategy is presented in Supplementary Fig. 1.

Jung H.E., Ku K.B., Kang B.H., Park J.H., Kim H.C., Kim K.D, & Lee H.K. (2023). Intranasal delivery of an adenovirus-vector vaccine co-expressing a modified spike protein and a genetic adjuvant confers lasting mucosal immunity against SARS-CoV-2. Antiviral Research, 216, 105656.

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Effector Memory T Cell Analysis

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Spleens were harvested from mice treated by a single i.t. injection of Saline or Mito‐FFa (60 µg per mouse) on Day 40. Use the flat end of the plunger to mince the spleen and then passed strainer. Red blood cells were removed by RBC Lysis and a single cell suspension were prepared. PE antimouse CD62L, FITC antimouse CD8a, APC antimouse CD44 antibodies (BioLegend, USA) were introduced to identify effector memory T cells (Tem, CD8⁺ CD62L⁻ CD44⁺) by flow cytometry (BD Calibur).

Wang Y., Wang W., Gu R., Chen J., Chen Q., Lin T., Wu J., Hu Y, & Yuan A. (2023). In Situ Vaccination with Mitochondria‐Targeting Immunogenic Death Inducer Elicits CD8+ T Cell‐Dependent Antitumor Immunity to Boost Tumor Immunotherapy. Advanced Science, 10(20), 2300286.

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Identification of T Cell Subsets

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PE anti-mouse CD3 (0.5 µg per million cells in 100 µL volume), FITC anti-mouse CD8a (1.0 µg per million cells in 100 µL volume), APC anti-mouse CD44 (0.25 µg per million cells in 100 µL volume) and PerCP/Cy5.5 anti-mouse CD62L (0.25 µg per million cells in 100 µL volume) antibodies (BioLegend, USA) were introduced to identify effector memory T cells (T_EM, CD3⁺ CD8⁺ CD62L⁻ CD44⁺) and central memory T cells (T_CM, CD3⁺ CD8⁺ CD62L⁺ CD44⁺) by flow cytometry (BD Calibur).

Huang Z., Wang Y., Yao D., Wu J., Hu Y, & Yuan A. (2021). Nanoscale coordination polymers induce immunogenic cell death by amplifying radiation therapy mediated oxidative stress. Nature Communications, 12, 145.

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