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Cxcl4

Manufactured by Merck Group
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CXCL4 is a laboratory product manufactured by Merck Group. It is a chemokine protein that plays a key role in the regulation of various cellular processes. The core function of CXCL4 is to facilitate cell migration and modulate immune system responses. Further details on its intended use are not available.

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Lab products found in correlation

Heparin, by Merck Group (2 mentions) Stemspan medium, by STEMCELL (2 mentions) Recombinant murine cxcl4, by Thermo Fisher Scientific (2 mentions) Micro slide, by Ibidi (1 mentions) Axiovert 200 microscope, by Zeiss (1 mentions) Rpmi media, by Merck Group (1 mentions) Rpmi medium, by Merck Group (1 mentions) Chromogenic lal endotoxin assay kit, by GenScript (1 mentions) Orn8l, by Chemgenes (1 mentions) Orn8l, by GenScript (1 mentions) Anti cd14 magnetic bead, by Miltenyi Biotec (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)

4 protocols using cxcl4

1

Monocyte-Derived Macrophage Migration Assay

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Microslides (Ibidi, Martinsried, Germany) were coated with collagen according to the manufacturer’s instructions; collagen was solidified by incubation at 37°C for 30 minutes. The first well of each capillary had peripheral blood monocyte–derived macrophages (PBMCs) in RPMI media (Sigma-Aldrich). The connecting well had 20 ng/mL CXCL4 [Sigma-Aldrich; half maximal effective concentration for CXCL4 as found by Baltus et al. in 2005 (11 (link))] in RPMI media; RPMI medium alone was used as a negative control. Movement of PBMCs was measured after 24 hours by using an Axiovert 200 microscope (Zeiss). Distance measured was converted into percentage movement, wherein complete movement would be 100% and no movement, 0%. The experiment was repeated with PBMCs from five different women.
Maybin J.A., Thiruchelvam U., Madhra M., Saunders P.T, & Critchley H.O. (2017). Steroids Regulate CXCL4 in the Human Endometrium During Menstruation to Enable Efficient Endometrial Repair. The Journal of Clinical Endocrinology and Metabolism, 102(6), 1851-1860.
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2

Murine Hematopoietic Stem Cell Proliferation

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For in vitro analyses of HSCs, lineage-depleted BMNCs were cultured for 7 days in StemSpan medium (StemCell Technologies) supplemented with Kitl (10 ng ml−1), Fgf1 (10 ng ml−1) and Thpo (20 ng ml−1; all R&D Systems). Recombinant murine Cxcl4 (100 ng ml−1; PeproTech) or vehicle with or without heparin (30 μg ml−1; Sigma) was added to assess Cxcl4-related effects on HSCs. Proliferation of HSCs was determined by adding 10 μl of a 1 mM BrdU solution to the cell cultures, staining and analysis was performed as described above.
Bruns I., Lucas D., Pinho S., Ahmed J., Lambert M.P., Kunisaki Y., Scheiermann C., Schiff L., Poncz M., Bergman A, & Frenette P.S. (2014). Megakaryocytes regulate hematopoietic stem cell quiescence via Cxcl4 secretion. Nature medicine, 20(11), 1315-1320.
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3

Murine Hematopoietic Stem Cell Proliferation

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For in vitro analyses of HSCs, lineage-depleted BMNCs were cultured for 7 days in StemSpan medium (StemCell Technologies) supplemented with Kitl (10 ng ml−1), Fgf1 (10 ng ml−1) and Thpo (20 ng ml−1; all R&D Systems). Recombinant murine Cxcl4 (100 ng ml−1; PeproTech) or vehicle with or without heparin (30 μg ml−1; Sigma) was added to assess Cxcl4-related effects on HSCs. Proliferation of HSCs was determined by adding 10 μl of a 1 mM BrdU solution to the cell cultures, staining and analysis was performed as described above.
Bruns I., Lucas D., Pinho S., Ahmed J., Lambert M.P., Kunisaki Y., Scheiermann C., Schiff L., Poncz M., Bergman A, & Frenette P.S. (2014). Megakaryocytes regulate hematopoietic stem cell quiescence via Cxcl4 secretion. Nature medicine, 20(11), 1315-1320.
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4

Monocyte isolation and stimulation

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Deidentified human buffy coats were purchased from the New York Blood Center following a protocol approved by the Hospital for Special Surgery Institutional Review Board. Peripheral blood mononuclear cells were isolated with Lymphoprep (Accurate Chemical) via density gradient centrifugation and monocytes were purified from peripheral blood mononuclear cells with anti-CD14 magnetic beads as recommended by the manufacturer (Miltenyi Biotec).10 (link) Monocytes were cultured overnight at 37 °C, 5% CO2 in RPMI-1640 medium (Invitrogen) supplemented with 10% heat-inactivated defined fetal bovine serum (FBS) (HyClone; Fisher Scientific), penicillin-streptomycin (Invitrogen), L-glutamine (Invitrogen), and 20 ng/mL human macrophage colony-stimulating factor (M-CSF). After at least 12-h culture, the cells were treated as described in the figure legends and were harvested and prepared for total RNA extraction, protein extraction, and flow cytometry. Cells were stimulated with CXCL4 (Sigma) and/or TLR8 ligand ORN8L (ChemGenes) as previously described10 (link); testing of various lots of CXCL4 using the Chromogenic LAL Endotoxin Assay Kit (Genscript) showed that CXCL4 preparations contributed 0.01 to 0.06 EU/mL final concentration of endotoxin in cultures, which does not contribute appreciably to synergy with ORN8L.10 (link)
Curnow A.W., Tumbula D.L., Pelaschier J.T., Min B, & Söll D. (1998). Glutamyl-tRNAGln amidotransferase in Deinococcus radiodurans may be confined to asparagine biosynthesis. Proceedings of the National Academy of Sciences of the United States of America, 95(22), 12838-12843.
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