A549, H460, H661, and H1299 lung cancer cell lines were obtained from the American Type Culture Collection (ATCC) (Manassas, VA, USA) and cultured with RPMI-1640 medium (Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Hyclone, South Logan, UT, USA). Cisplatin, 4′,6-diamidino-2-phenylindole (DAPI), and a PKH-26 Red Fluorescent Cell Linker Kit were purchased from Sigma-Aldrich Chemical Company (St. Louis, MO, USA). FITC-conjugated CD44 antibody, APC-conjugated CD90 antibody, and corresponding isotype controls were purchased from BD Biosciences (San Jose, CA, USA). Anti-human CD44, CD90, and p53 primary antibodies were purchased from Abcam (Cambridge, MA, USA). CCR2 antibody was obtained from R&D (Minneapolis, MN, USA); CD68 antibody and Lipofectamine 2000 reagent were purchased from Invitrogen (Carlsbad, CA, USA). Secondary antibodies FITC-conjugated goat anti-rabbit IgG and Cy3-conjugated goat anti-mouse IgG were purchased from Jackson ImmunoResearch (West Grove, PA, USA). pCMV-Neo-Bam p53 R249S was a gift from Bert Vogelstein (Addgene plasmid # 16438), and pCMV-Neo-Bam p53 wt (wt p53) was a gift from Bert Vogelstein (Addgene plasmid # 16434).
Fitc conjugated cd44 antibody
Manufactured by BD
Sourced in United States
The FITC-conjugated CD44 antibody is a laboratory reagent used for the detection and analysis of CD44 protein expression in biological samples. CD44 is a cell surface glycoprotein involved in cell-cell interactions, cell adhesion, and migration. The FITC (Fluorescein Isothiocyanate) conjugation allows for the visualization and quantification of CD44-positive cells using flow cytometry or fluorescence microscopy techniques.
Automatically generated - may contain errors
Lab products found in correlation
Magnetic cell sorting system, by Miltenyi Biotec (2 mentions)
Accutase, by Innovative Cell Technologies (2 mentions)
Facscalibur, by BD (2 mentions)
Pkh26 red fluorescent cell linker kit, by Merck Group (1 mentions)
Apc conjugated cd90 antibody, by BD (1 mentions)
Fitc conjugated goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Cy3 conjugated goat anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Pcmv neo bam p53 r249s, by Addgene (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Cisplatin, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
H1299, by American Type Culture Collection (1 mentions)
Ab51037, by Abcam (1 mentions)
Ab32072, by Abcam (1 mentions)
Bd555742, by BD (1 mentions)
Fitc conjugated cd44, by BD (1 mentions)
Pe conjugated cd133 antibody, by BioLegend (1 mentions)
Aldefluor stem cell identification kit, by STEMCELL (1 mentions)
Cd133 microbead kit, by Miltenyi Biotec (1 mentions)
5 protocols using fitc conjugated cd44 antibody
1
Lung Cancer Cell Line Characterization
2
Evaluating Anticancer Treatments via Flow Cytometry
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Antibodies against CD44 (Abcam, Cat #ab51037), c-Myc (Abcam, Cat #ab32072), CBFb (Abcam, Cat #ab133600), and GAPDH (ZSGB-BIO, Cat #TA-08) were obtained commercially. FITC-conjugated CD44 antibody (BD, Cat #560977) was purchased from BD Biosciences (Franklin Lakes, NJ, USA). Statins were purchased from Abcam (atorvastatin [ab145604], lovastatin [ab120614], and simvastatin [ab120505]) or Sigma-Aldrich (St Louis, MO, USA) and dissolved in DMSO as a 20 ~ 30 mM stock solution and stored at -20°C before use. Chemotherapeutic drugs (cyclophosphamide [CPA], cisplatin [DDP], doxorubicin [DOX], and paclitaxel [TXL] were obtained from the in-hospital pharmacy or Selleck (Shanghai, China). The photosensitizer polyhematoporphyrin (C34H38N4NaO5, Photosan-II [PS-II]) was purchased from Seehof Laboratorium F&E GmbH (Wesselburenerkoog, Germany).
3
Cell Sorting and Quantification via CD44
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For cell sorting, cells were dissociated using Accutase (Innovative Cell Technologies). For FACS, cells were resuspended in PBS containing 0.5% BSA, stained with FITC-conjugated CD44 (BD555478) or isotype control antibody (BD555742) from BD Biosciences and analysed on a BD FACSCalibur (BD Biosciences) using Cell Quest software. CD44-positive cells were sorted using a magnetic cell sorting system (Miltenyi Biotech). Cells were stained with CD44 Micro-Beads, passed through a LS magnetic column, then eluted from the column after removal from the magnet. CD44-positive cells were quantified by immunofluorescence using FITC-conjugated CD44 antibody (555478; BD Biosciences).
4
Prostate Cancer Stem Cells Characterization
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For PCSCs population analysis, prostate cancer cells were incubated with FITC-conjugated CD44 antibody (11-0441-82, BD Biosciences, San Diego, CA, USA) and PE-conjugated CD133 antibody (372804, Biolegend, San Diego, CA, USA) for 30 min at 37 °C. The CD44+/CD133+ subpopulation was quantified by flow cytometry and defined as PCSCs. Hyperactive aldehyde dehydrogenase (ALDH) activity is closely related to the physiological properties of CSCs50 (link). PCSCs population analysis was also conducted by flow cytometry using the ALDEFLUOR Stem Cell Identification Kit (01700, STEMCELL Technologies, Vancouver, Canada) according to the manufacturer’s instructions. The ALDH+ subpopulation was quantified and defined as PCSCs. As a specific inhibitor of ALDH activity, diethylaminobenzaldehyde (DEAB), was used to control for background fluorescence in the ALDH staining assay. For PCSCs sorting, the CD133+ subpopulation was isolated by the magnetic activated cell sorting (MACS) method using the CD133 MicroBead Kit (130-100-857, Miltenyi, North Rhine-Westphalia, Germany) in accordance with the manufacturer’s instructions.
5
Isolation and Characterization of CD44+ Cells
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