Bx471

Manufactured by Merck Group

Sourced in United Kingdom

BX471 is a laboratory equipment product manufactured by Merck Group. It is a device designed for general laboratory applications. The core function of BX471 is to perform basic laboratory tasks, but a detailed description cannot be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

Sb225002, by Merck Group (1 mentions) Amg487, by Merck Group (1 mentions) Cell counting kit 8 cck 8, by Absin (1 mentions) Bx471, by Bio-Techne (1 mentions) Maraviroc, by Apexbio (1 mentions) Il 10 neutralizing antibody, by R&D Systems (1 mentions) Chemotx, by Neuro Probe (1 mentions) Fluorescent beads, by BD (1 mentions) Maraviroc, by Merck Group (1 mentions) Classical monocyte isolation kit, by Miltenyi Biotec (1 mentions) Control goat igg, by Merck Group (1 mentions) Matrigel, by BD (1 mentions) Accuri c6, by BD (1 mentions) Mycoalert plus mycoplasma detection kit, by Lonza (1 mentions) Caerulein, by Merck Group (1 mentions) Diphtheria toxin, by Enzo Life Sciences (1 mentions)

6 protocols using bx471

Chemokine Receptor Antagonist Effects on RA-FLS Viability

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RA-FLS were seeded into 96-well plates at a density of 5x10³ cells/well for 24 h and subsequently treated with different concentrations of several chemokine receptor antagonists: BX471 (50, 100 and 150 nM; CCR1 antagonist, Sigma-Aldrich; Merck KGaA), AMG487 (0.5, 1 and 2 µM; CXCR3 antagonist, Sigma-Aldrich; Merck KGaA) and SB225002 (0.1, 0.2 and 0.4 µM; CXCR2 antagonist, Sigma-Aldrich; Merck KGaA). Following incubation for 24 h at 37˚C, 10 µl Cell Counting Kit-8 (CCK-8; Absin Bioscience, Inc.) reagent was added to each well and the optical density was measured at a wavelength of 450 nm.

Wang W., Deng Z., Liu G., Yang J., Zhou W., Zhang C., Shen W, & Zhang Y. (2021). Platelet-derived extracellular vesicles promote the migration and invasion of rheumatoid arthritis fibroblast-like synoviocytes via CXCR2 signaling. Experimental and Therapeutic Medicine, 22(4), 1120.

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Subcutaneous CCR1 Antagonist Treatment Post-Photic Injury

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Immediately after photic injury was induced, the mice received subcutaneous injections of either the CCR1-specific antagonist BX471 (50 mg/kg body weight; Tocris, Bristol, UK) or vehicle (40% cyclodextrin in saline) every 12 hr for 5 days. For injection, BX471 was dissolved at a final concentration of 10 mg/ml in saline containing 40% (wt/vol) cyclodextrin (Sigma-Aldrich); the solution was mixed thoroughly and dissolved overnight at 4°C, after which the pH was adjusted to 4.5 with NaOH, and the solution was filtered through a 0.45 µm filter.

Elbaz-Hayoun S., Rinsky B., Hagbi-Levi S., Grunin M, & Chowers I. (2023). CCR1 mediates Müller cell activation and photoreceptor cell death in macular and retinal degeneration. eLife, 12, e81208.

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Chemotaxis Assay of Monocyte Migration

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Conditioned or control media were diluted 1:2 in chemotaxis medium (PBS with 1% albumin, low endotoxin; Sigma). Monocytes were freshly isolated from HDs after obtaining written informed consent using negative MACS depletion as described previously^{20 (link)} and resuspended in chemotaxis medium. The diluted media were added in the lower chambers of a 5 μm chemotaxis assay plate (96 well ChemoTX®, NeuroProbe, Gaithersburg, MA, USA) and 100,000 monocytes were transferred to the upper chamber. After 2 h, chemotaxis was quantified by measuring the DAPI (4,6 diamidino-2-phenylindole) signal of migrated monocytes as described previously.^{20 (link)}When measuring inhibitor effects, both media and monocytes were incubated for 30 minutes (min) on ice with the indicated inhibitors directly prior to the migration assay. The following chemokine receptor inhibitors were used: 1 μg/mL CCR1 inhibitor BX471 (Sigma), 1 μg/mL CCR2 inhibitor INCB3284 (Tocris Bioscience, Bristol, UK), 1μM CCR3 inhibitor SB328437 (Tocris), 1 μM CCR5 inhibitor Maraviroc (Apexbio, Houston, TX, USA), 1μM CXCR4/7 inhibitor Plerixafor (Apexbio), and 0.1 μg/mL IL-10 neutralizing antibody (R&D Systems, Minneapolis, MN, USA).

van Attekum M.H., van Bruggen J.A., Slinger E., Lebre M.C., Reinen E., Kersting S., Eldering E, & Kater A.P. (2017). CD40 signaling instructs chronic lymphocytic leukemia cells to attract monocytes via the CCR2 axis. Haematologica, 102(12), 2069-2076.

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Monocyte Invasion Assay with DLBCL

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Monocytes were purified by negative selection with a Classical Monocyte Isolation Kit (Miltenyi Biotec), according to the manufacturer’s instructions. Purity exceeded 80%. DLBCL cells were incubated in bottom compartments of 8-µM 24-well Transwell plates (Nunc) at 0.5 × 10⁶ cells per milliliter. Twenty-four hours later, 0.2 × 10⁶ purified monocytes were added to upper compartments. After 2 hours of incubation, cells from the bottom compartment were stained for CD14, and CD14⁺ monocytes were enumerated by flow cytometry with an Accuri C6 (BD Biosciences) set on a volume-based event acquisition. Flow variations were controlled by the addition of fluorescent beads (BD Biosciences). The invasion assay was performed similarly but the filter was covered with 50 µL of Matrigel (BD Biosciences), and the incubation time was extended to 18 hours. Polyclonal goat anti-CCL5 (R&D Systems) and control goat IgG (Sigma) were used at 10 µg/mL. metCCL5 was used at 100 ng/mL. Maraviroc and BX471 (Sigma) were used at 100 µM and 100 nM, respectively.

Manfroi B., De Grandis M., Moreaux J., Tabruyn S., Mayol J.F., Quintero M., Righini C., Sturm N., Aurrand-Lions M, & Huard B. (2021). The microenvironment of DLBCL is characterized by noncanonical macrophages recruited by tumor-derived CCL5. Blood Advances, 5(21), 4338-4351.

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Dual-Inhibitor Therapy for bcCML

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MVC (Selleck Chemicals) was prepared in 100% ethanol and diluted to a 5 mg/mL working solution in sterile PBS immediately prior to injection. Mice were treated with a 20 mg/kg dose of MVC by intraperitoneal injection twice daily with injections given 6 hours apart for 9 days, 1 day after bcCML initiation. BX471 (Sigma) was prepared in 40% w/v cyclodextrin and diluted to a 2.5 mg/mL working solution in sterile saline immediately prior to injection. Mice were treated with a 10 mg/kg dose of BX471 by intraperitoneal injection twice daily with injections given 6 hours apart for 9 days, 1 day after bcCML initiation. Mice were sacrificed on day 11 for flow cytometry analysis.

Ackun-Farmmer M.A., Soto C.A., Lesch M.L., Byun D., Yang L., Calvi L.M., Benoit D.S, & Frisch B.J. (2021). Reduction of leukemic burden via bone-targeted nanoparticle delivery of an inhibitor of C-chemokine (C-C motif) ligand 3 (CCL3) signaling. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 35(4), e21402.

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Induction and Depletion of Regulatory T Cells in Pancreatic Cancer

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For Treg depletion, Foxp3^DTR, KC;Foxp3^DTR mice were treated with diphtheria toxin (DT) (50 ng/g) (Enzo Life Science) by intraperitoneal injection (i.p.). DT injections were repeated according to the specific experimental design shown in Figures. Control mice lacking Foxp3^DTR alleles received the same DT treatments. In KC, KC;Foxp3^DTR and KC;CD4^−/− mice, mild acute pancreatitis was induced in 4–5-week-old mice by a series of 8 hourly intraperitoneal injections of caerulein (Sigma-Aldrich, 75 μg/kg) over 1-day period. For CCR1 inhibition, mice were subcutaneously dosed with CCR1 antagonist BX471 (Sigma-Aldrich, 50 mg/kg) for 7 days at 12-hour intervals. DMSO was used as vehicle to dissolve BX471 at a concentration of 50 mg/ml.
To establish the orthotopic pancreatic cancer model, 1×10⁵ of 7940B cells (C57BL/6J strain)(46 (link)) derived from KPC tumor (Ptf1a-Cre; LSL-Kras^G12D; p53^flox/+) were injected into Foxp3^DTR mice of compatible genetic background. Cells were tested for mycoplasma free by MycoAlertTM PLUS Mycoplasma Detection Kit (Lonza) and passage 15–20 were used for all experiments. For CD8⁺ T cell depletion, anti-CD8 mAb (BioXcell clone 2.43; 200 μg/mouse) was injected i.p. twice per week. For CD4⁺ T cell depletion, anti-CD4 mAb (BioXcell clone GK1.5; 200 μg/mouse) was injected i.p. at least every three days.

Zhang Y., Lazarus J., Steele N.G., Yan W., Lee H.J., Nwosu Z.C., Halbrook C.J., Menjivar R.E., Kemp S.B., Sirihorachai V.R., Velez-Delgado A., Donahue K., Carpenter E.S., Brown K.L., Irizarry-Negron V., Nevison A.C., Vinta A., Anderson M.A., Crawford H.C., Lyssiotis C.A., Frankel T.L., Bednar F, & Pasca di Magliano M. (2020). Regulatory T cell depletion alters the tumor microenvironment and accelerates pancreatic carcinogenesis.. Cancer discovery, 10(3), 422-439.

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