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Apc anti cd44 antibody

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The APC-anti-CD44 antibody is a fluorescent-labeled antibody that binds to the CD44 cell surface protein. CD44 is a receptor for hyaluronic acid and is involved in cell-cell interactions, cell adhesion, and migration. The APC-anti-CD44 antibody can be used to identify and quantify CD44-expressing cells in flow cytometry applications.

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Lab products found in correlation

Facs aria high speed cell sorter, by BD (3 mentions) Facscanto 2, by BD (2 mentions) Facsaria dual laser flow cytometer, by BD (1 mentions) Pe anti human cd184 cxcr4 antibody, by BioLegend (1 mentions) Pe annexin 5 apoptosis detection kit 1, by BD (1 mentions) Facsariaii high speed cell sorter, by BD (1 mentions) Facsaria iiu flow cytometer, by BD (1 mentions) Fitc anti human cd326 epcam antibody, by BioLegend (1 mentions) Aria sorter, by BD (1 mentions) Aldefluor kit, by STEMCELL (1 mentions) Cytometric bead array, by BD (1 mentions) Anti cd62l fitc antibody, by BD (1 mentions) Mouse th1 th2 th17 cytokine kit, by BD (1 mentions) Anti cd8 pe, by BD (1 mentions) Anti cd3, by BD (1 mentions) Anti cd28, by BD (1 mentions) Lsrfortessa cell analyzer, by BD (1 mentions) Aldefluortm kit, by STEMCELL (1 mentions) Transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions) Diva software, by BD (1 mentions)

9 protocols using apc anti cd44 antibody

1

Quantifying CD133+/CD44+ Prostate Cancer Cells

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Control PCa cells (PC3Control or DU145Control) or GAS6 overexpressed PCa cells (PC3GAS6OE or DU145GAS6OE) (1 × 105) were seeded onto 12-well culture plates, and then cultured for 48 hours. PCa cells were incubated with PE-anti-CD133 antibody (cat. 130-080-901, Miltenyi Biotec, San Diego, CA) and APC-anti-CD44 antibody (cat. 559942, BD Biosciences, San Jose, CA) for 20 min at 4°C. The CD133+/CD44+ fraction was analyzed with a FACS Aria High-Speed Cell Sorter (BD Biosciences).
Jung Y., Decker A.M., Wang J., Lee E., Kana L.A., Yumoto K., Cackowski F.C., Rhee J., Carmeliet P., Buttitta L., Morgan T.M, & Taichman R.S. (2016). Endogenous GAS6 and Mer receptor signaling regulate prostate cancer stem cells in bone marrow. Oncotarget, 7(18), 25698-25711.
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2

Analyzing Cancer Stem Cell Phenotype in Prostate Cancer

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For analysis of a cancer stem cell phenotype (CD133+/CD44+), overexpression of CXCL12γ in PCa or control cells (PC3, DU145) (1 × 105) were seeded onto 12-well culture plates and were cultured for 4 days. The cells were incubated with PE-anti-CD133 antibody (cat. 130–080-901, Miltenyi Biotec, San Diego, CA) and APC-anti-CD44 antibody (cat. 559942, BD Biosciences, San Jose, CA) for 20 min at 4°C. For CXCR4 positive cell analysis, the cells were incubated with PE-anti-human CD184 (CXCR4) antibody (cat. 306506, BioLegend, San Diego, CA) or mouse IgG-PE (cat. 130–092-212, Miltenyi Biotec) for 20 min at 4°C. The CD133+/CD44+ or CXCR4 positive fractions were analyzed with a FACS Aria High-Speed Cell Sorter (BD Biosciences). Apoptosis was measured by flow cytometry (FACSAria dual laser flow-cytometer, Becton Dickinson, Mountainview, CA) using PE Annexin V Apoptosis Detection Kit I (cat. 559763, BD Biosciences, San Jose, CA). The PCa cells were pretreated AMD3100 (5μg/ml) or siCXCR4 and treated with of docetaxel (Taxotere; 0.5–1μg/ml, Hospira, Lake Forest, IL). In some cases, the PCa cells were treated with XTANDI® (enzalutamide; 0.5μg/ml)‎ (Selleck Chemicals, Houston, TX).
Jung Y., Cackowski F.C., Yumoto K., Decker A.M., Wang J., Kim J.K., Lee E., Wang Y., Chung J.S., Gursky A.M., Krebsbach P.H., Pienta K.J., Morgan T.M, & Taichman R.S. (2018). CXCL12γ Promotes Development of Metastatic Castration Resistant Prostate Cancer by Induction of Cancer Stem Cell and Neuroendocrine Phenotypes. Cancer research, 78(8), 2026-2039.
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3

Multiparameter Flow Cytometry of Cell Lines

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Cells were stained with FITC- or APC-anti-HLA-ABC antibody (clone W6/32: 311404 (FITC), 311410 (APC), BioLegend, San Diego, CA), PE-cytokeratin antibody (347204, Becton Dickinson, Franklin Lakes, NJ), PE-EpCAM antibody (130-091-253, Miltenyi Biotec), PE-anti-CD133 antibody (130-080-801, Miltenyi Biotec), APC-anti-CD44 antibody (559942, BD Biosciences, San Diego, CA), or isotype-matched IgG control for 20 min at 4°C. Flow cytometric analyses were performed in a FACSAria II High-Speed Cell Sorter (Becton Dickinson).
Shiozawa Y., Berry J.E., Eber M.R., Jung Y., Yumoto K., Cackowski F.C., Yoon H.J., Parsana P., Mehra R., Wang J., McGee S., Lee E., Nagrath S., Pienta K.J, & Taichman R.S. (2016). The marrow niche controls the cancer stem cell phenotype of disseminated prostate cancer. Oncotarget, 7(27), 41217-41232.
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4

Characterization of CD133+/CD44+ Prostate Cancer Cells

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PCa cells (PC3 or DU145) (1 × 105) were seeded onto 12-well culture plates. The cells were treated with vehicle or a demethylating agent, 5-Aza (0-20 μM) for 4 days. PCa cells were incubated with PE-anti-CD133 antibody (cat. 130-080-901; Miltenyi Biotec, San Diego, CA) and APC-anti-CD44 antibody (cat. 559942; BD Biosciences, San Jose, CA) for 20 minutes at 4°C. The CD133+/CD44+ fraction was analyzed with a FACS Aria High-Speed Cell Sorter (BD Biosciences).
Lee E., Wang J., Yumoto K., Jung Y., Cackowski F.C., Decker A.M., Li Y., Franceschi R.T., Pienta K.J, & Taichman R.S. (2016). DNMT1 Regulates Epithelial-Mesenchymal Transition and Cancer Stem Cells, Which Promotes Prostate Cancer Metastasis. Neoplasia (New York, N.Y.), 18(9), 553-566.
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5

Characterization of Epithelial Cell Phenotypes

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For analyses of epithelial cell phenotypes, luminal (EPCAMHi/CD49fLo) or non-luminal (EPCAMLoCD49fHi) and an epithelial stem cell phenotype (EPCAMLo/CD49fHi/CD24Lo/CD44Hi)8 (link), 48 , 49 (link), control or CXCL12γ overexpressing epithelial cells (PNT2, MCF10A) (1 x 105) were seeded onto 12-well culture plates and were cultured for 3 days. The cells were incubated with FITC anti-human CD326 (EPCAM) antibody (cat. 324204, Biolegend, San Diego, CA), PE/Cy7 anti-human CD49f antibody (cat. 313622, Biolegend), APC/Cy7 anti-human CD24 antibody (cat. 311132, Biolegend), APC anti-CD44 antibody (cat. 559942, BD Biosciences, San Jose, CA) for 30 min at 4°C. DAPI was used for determining cell viability. The cell phenotypes were evaluated by FACS analyses (FACSAria IIu flow-cytometer, Becton Dickinson, Mountain View, CA). Assays were performed in triplicate and the results are representative of three independent experiments.
Jung Y., Kim J.K., Lee E., Cackowski F.C., Decker A.M., Krebsbach P.H, & Taichman R.S. (2020). CXCL12γ Induces Human Prostate and Mammary Gland Development. The Prostate, 80(13), 1145-1156.
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6

ALDH Activity and CD44 Expression Analysis

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ALDH activity was detected by ALDEFLUOR kit (Stem Cell Technologies) according to the manufacturer's instructions. CD44 expression was stained by anti‐CD44‐APC antibody (BD Biosciences) as previously described.[15a] Cells were then analyzed using FACS Canto II (BD Biosciences). Results were analyzed using FlowJo software (Tree star). Cell isolation and enrichment were performed using BD Aria sorter (BD Biosciences). The purity of sorted cells was at least 95%.
Wang S., Chen J., Jiang Y., Lei Z., Ruan Y.C., Pan Y., Yam J.W., Wong M.P, & Xiao Z. (2023). Targeting GSTP1 as Therapeutic Strategy against Lung Adenocarcinoma Stemness and Resistance to Tyrosine Kinase Inhibitors. Advanced Science, 10(7), 2205262.
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7

Profiling Lymphocyte Responses in Mice

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Nine days after the protein boost (on day 21 after birth), the animals were euthanized, and the spleens and lungs were collected aseptically. The cell preparations were conducted as previously described [26 (link)]. The cell suspensions were adjusted to 1 × 106 cells/mL and incubated with anti-CD28 (1 µg/mL, clone: CD82.2, BD Pharmingen™), anti-CD3 (1 µg/mL, clone: OKT3, BD Pharmingen™), and rPspA-PdT (5 µg/mL) for 48 h at 37 °C and 5% CO2. The supernatant sample was collected, and the concentrations of cytokines (IFN-γ, IL-17, TNF-α, IL-10, and IL-6) were evaluated via Cytometric Bead Array (BD Pharmingen™) using the Mouse Th1/Th2/Th17 Cytokine Kit.
Additionally, to investigate the phenotype of memory B and T cells in the lungs and spleen, the remaining cells were labeled with specific antibodies: (i) Memory B cells: anti-B220-FITC antibody (clone: RA3-6B2, BD Pharmingen™), anti-CD19-BV421 antibody (clone: 1D3, BD Horizon™), and anti-CD27-PerCP.Cy5.5 antibody (clone: LG.3A10, BD Pharmingen™); (ii) Memory T cells: anti-CD4-APC.Cy7 antibody (clone: GK1.5, BD Pharmingen™), anti-CD8-PE.Cy7 antibody (clone:53-6.7, BD Pharmingen™), anti-CD62L-FITC antibody (clone: MEL-14, BD Pharmingen™), and anti-CD44-APC antibody (clone: IM7, BD Pharmingen™) for 30 min. Also, the cells were rinsed and resuspended in 1% PFA. Data were acquired using a FACSCanto II flow cytometer.
Trentini M.M., Rodriguez D., Kanno A.I., Goulart C., Darrieux M, & de Cerqueira Leite L.C. (2024). Robust Immune Response and Protection against Lethal Pneumococcal Challenge with a Recombinant BCG-PspA-PdT Prime/Boost Scheme Administered to Neonatal Mice. Vaccines, 12(2), 122.
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8

Multiparameter Flow Cytometry Analysis of Immune Markers

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For ALDH/CD44 staining, ALDH activity was detected with an ALDEFLUORTM kit (Stem Cell Technologies) according to the manufacturer's instructions. CD44 expression was detected with an anti‐CD44‐APC antibody (BD Biosciences) as previously described.[18 (link)
] For the analysis of cell surface proteins, the cells were incubated with antibodies in a staining wash buffer for 30 min in the dark. For intracellular proteins (FOXP3 and IFNγ), cells were fixed, permeabilized, and stained using a Transcription Factor Staining Buffer Set (eBioscience). The cells were then washed and resuspended in 0.5 mL of staining buffer. The antibodies used included APC‐anti‐human CD3, PE‐anti‐human CD4, FITC‐anti‐human CD8, PE‐anti‐human IFNγ, FITC‐anti‐human FOXP3, BV785‐anti‐human PD‐1, BV605‐anti‐human TIM3, APC/CY7‐anti‐mouse CD45, Percp5.5‐anti‐mouse CD3, FITC‐anti‐mouse CD4, PE/CY7‐anti‐mouse CD8a, and pacific blue‐anti‐mouse FOXP3, Brilliant Violet 785 anti‐mouse/human CD44, Brilliant Violet 605 anti‐mouse CD279 (PD‐1), PE/Cyanine7 anti‐mouse CD366 (Tim‐3), PE anti‐mouse CD62L, APC/Fire 750 anti‐mouse CD3, Brilliant Violet 510 anti‐mouse CD8a, Alexa Fluor 700 anti‐mouse CD45. All antibodies used were purchased from Biolegend. The cells were analyzed using a BD LSRFortessa Cell Analyzer (BD Biosciences). The results were analyzed using FlowJo software (Tree Star).
Xiao Z., Wang S., Chen J., Li Y., Jiang Y., Tin V.P., Liu J., Hu H., Wong M.P., Pan Y, & Yam J.W. (2024). The Dual Role of the NFATc2/galectin‐9 Axis in Modulating Tumor‐Initiating Cell Phenotypes and Immune Suppression in Lung Adenocarcinoma. Advanced Science, 11(20), 2306059.
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9

Autophagy Flux Measurement Protocol

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After infections and/or treatments, cells were resuspended in PBS with 0.5% bovine serum albumin and 2 mM EDTA buffer. Autophagic flux was directly measured as previously described [31, 32] . The appropriate forward/side scatter profile was used to exclude non-viable cells. All mCherry-positive cells were included in the gate to measure the mCherry/ GFP ratio, with a scaling of 25%. For CD44 quantification, cells were stained with anti-CD44-APC antibody (1:25 each, BD Pharmingen, San Jose, CA, USA). Flow cytometry experiments were performed using BD LRSFortessa and BD FACS CantoII instruments, and DIVA software (both from Becton Dickinson, Le Pont de Claix, France). Results were based on the analysis of 10,000 cells.
Courtois S., Haykal M., Bodineau C., Sifré E., Azzi-Martin L., Ménard A., Mégraud F., Lehours P., Durán R.V., Varon C, & Bessède E. (2021). Autophagy induced by Helicobacter pylori infection is necessary for gastric cancer stem cell emergence. Gastric cancer : official journal of the International Gastric Cancer Association and the Japanese Gastric Cancer Association, 24(1).
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