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Anti mouse cd11b fitc

Manufactured by Miltenyi Biotec
Sourced in Italy, United Kingdom

Anti-mouse CD11b FITC is a fluorescently-labeled monoclonal antibody that specifically binds to the CD11b cell surface antigen expressed on various murine immune cells, including monocytes, macrophages, and granulocytes. It can be used for the identification and analysis of these cell populations in flow cytometry applications.

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2 protocols using anti mouse cd11b fitc

1

Isolation of Tumor-Infiltrating Leukocytes from Mouse Models

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Six to eight mm mean diameter tumors from neuT and neuT-pfpKO mice were minced with scalpels and then incubated with 1 mg/mL collagenase IV (Sigma-Aldrich) in RPMI-1640 (Life Technologies, Monza, Italy) at 37 °C for 1 h in an orbital shaker. The cell suspension was washed in PBS supplemented with 2% FBS (Invitrogen), incubated in a buffer for erythrocyte lysis (155 mM NH4Cl, 15.8 mM Na2CO3, 1 mM EDTA, pH 7.3) for 10 min at RT, and then washed in RPMI-1640 supplemented with 10% FBS (Invitrogen). The cell suspension was passed through a 70-µm pore cell strainer (BD Biosciences, Milano, Italy), centrifuged (1400 rpm for 10 min), and re-suspended in a buffer for erythrocyte lysis. After 10 min of incubation at RT, tumor-infiltrating leukocytes were washed with RPMI-1640 supplemented with 10% FBS (Invitrogen), centrifuged (1400 rpm for 10 min), re-suspended in PBS, treated with Fc receptor blocker (anti CD16/CD32; BD Biosciences, Milano, Italy), and stained with the following antibodies: anti-mouse CD45 VioGreen, anti-mouse CD3 FITC, anti-mouse CD49b PE, anti-mouse CD4 APC Vio770, anti-mouse CD8 VioBlue, anti-mouse γδ TCR PE/Cy7, anti-mouse CD11b FITC, anti-mouse F4/80 APC, and anti-mouse GR-1 PE (Miltenyi Biotech, Milano, Italy) [27 (link)]. Samples were acquired and analyzed on a CyAn ADP (DakoCytomation, Milano, Italy) using Summit 4.3 software (DakoCytomation, Milano, Italy).
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2

BV2 Cell Activation by 225THC

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BV2 cells were treated with 1, 5, or 10 μM of 225THC for 2 hours prior to the addition of 2 μg/mL LPS for a further 48 hours. Cells were harvested by washing in Dulbecco's phosphate buffered saline (PBS, without Ca2+ and Mg2+) for 20 minutes at 37°C and then resuspended in cold PBS containing 0.5% BSA/0.05% NaN3 to metabolically fix the cells. Cells were stained with anti-mouse CD11b-FITC, CD40-FITC, CD54-FITC, and CD68-FITC, or appropriate FITC-conjugated isotype control antibodies (Miltenyi Biotec, UK) as per the manufacturer's instructions and cell surface staining was assessed by flow cytometry (FACSCalibur running CellQuest Pro; Becton Dickinson, UK) and analysed using Flowing software v2.5.1. Data are presented as average fold change in mean fluorescence intensity (MFI) versus untreated cells ± SEM from 4 independent experiments.
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