BHT101 cells, a kind gift of Prof. M. Santoro, Federico II University of Naples, were grown in Dulbecco's modified Eagle's medium (DMEM) (Sigma, St. Louis, MO USA) supplemented with 10% fetal bovine serum (Sigma). THP-1 cells were grown in RPMI 1640 medium (Sigma) supplemented with 10% fetal bovine serum (Sigma). To over-express D6/ACKR2 receptor, BHT101 cells were infected with pLenti-CMV-GFP-2A-Puro lentiviral vector (ABM, Richmond, BC, Canada) alone or containing the cDNA coding for human D6/ACKR2 fused in frame with FLAG tag. Anti-actin (sc-8432), anti-tubulin (sc-33749) and anti-IκBα (sc-371) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA), anti-FLAG (F7425 and F3165) were from Sigma, anti-D6/ACKR2 (human, PA5-11867) was from Pierce-Thermo Fisher Scientific (Waltham, MA USA), anti-F4/80 (MCA497R) was from AbD Serotec (Oxford, UK). Anti-PMCA (Plasma Membrane Calcium ATPase, clone 5F10) was from ABR Affinity BioReagents, Golden Co., USA. Anti-miR-146a was from Ambion-Thermo Fisher Scientific.
Anti f4 80 mca497r
Manufactured by Bio-Rad
Sourced in United States
The Anti-F4/80 (MCA497R) is a monoclonal antibody that recognizes the F4/80 antigen, a glycoprotein expressed on the surface of mature mouse macrophages. This antibody can be used for the identification and characterization of mouse macrophages in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti actin sc 8432, by Santa Cruz Biotechnology (1 mentions)
Anti iκbα sc 371, by Santa Cruz Biotechnology (1 mentions)
Plenti cmv gfp 2a puro lentiviral vector, by Applied Biological Materials (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
Np0335, by Thermo Fisher Scientific (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
3 protocols using anti f4 80 mca497r
1
Overexpression of D6/ACKR2 Receptor in BHT101 Cells
2
Immunoblotting of Insulin Receptor
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Proteins were extracted using CHAPS buffer (1.25% CHAPS; 28300; Thermo Fisher Scientific) with protease inhibitor cocktail (04693132001; Roche), separated on 4–20% SDS-PAGE (NP0335; Life Technologies), and transferred to nitrocellulose membranes (170-4158; Bio-Rad). Antibodies used were anti-F4/80 (MCA497R; AbD Serotec), anti–insulin receptor (IR; sc-711; Santa Cruz Biotechnology), actin (4970; Cell Signaling Technology), and secondary antibodies (7074; Cell Signaling Technology).
3
Histological Analysis of Hepatic Macrophages
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Tissues were fixed in 10% zinc formalin, then processed for paraffin sectioning (5-μm sections), and stained with hematoxylin and eosin. Anti-F4/80 (MCA497R; AbD Serotec) was used to assess macrophage infiltration in the liver.
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