The oligodeoxynucelotides AS1411, (5’-d(GGTGGTGGTGGTTGTGGTGGTGGTGG)-3’, and inactive control oligonucleotide CRO, 5’-d(CCTCCTCCTCCTTCTCCTCCTCCTCC)-3′ were synthesized by the Tufts Core DNA Synthesis facility (Tufts University, Boston, MA). AlexaFluro594 labeled AS1411, 5’-FL-AS1411, and AlexaFluro594 labeled CRO, 5’-FL-CRO, were purchased from Invitrogen and used for localization studies. All oligonucleotides were dissolved in nuclease free water. Neutralizing goat antibody against mouse VEGF-A (AF-493) was purchased from R&D Systems (Minneapolis, MN) and 2pmol were administered via intravitreal injection for lasered mice.
Af 493
Manufactured by R&D Systems
Sourced in United States
AF-493 is a recombinant human protein manufactured by R&D Systems. It is a member of the Activin A receptor type IB (ACVR1B) family.
Automatically generated - may contain errors
Lab products found in correlation
Ab34074, by Abcam (2 mentions)
Ab 108 c, by R&D Systems (2 mentions)
Af 233 na, by R&D Systems (1 mentions)
Matrigel matrix, by Corning (1 mentions)
0.4 μm transwell insert, by Corning (1 mentions)
Mab50201, by R&D Systems (1 mentions)
Mab6325, by R&D Systems (1 mentions)
Af791, by R&D Systems (1 mentions)
Sb431542, by Merck Group (1 mentions)
3 protocols using af 493
1
Oligonucleotides for cancer treatment
2
Tube Formation Assay of Endothelial Cells
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ECs were induced to form tubes according to our previously published methods [22 (link)]. Matrigel matrix (Corning) was added to 24-well culture plates (289 μL per well) on ice and then incubated at 37 °C for 1 h. ECs (12 × 104 cells in 300 μL per well) were seeded on polymerized Matrigel with 5 μM 152RM. After incubation on the angiogenesis assay plate for 3 h or 6 h at 37 °C, tube formation was observed with a microscope (Olympus, Tokyo, Japan), and cumulative tube length was measured. In some assays, neutralizing antibodies against VEGF (AF-493; R&D, Minneapolis, Minnesota, USA), PDGF-BB (ab34074; Abcam), TGF-β1 (9016; R&D), TGF-β3 (20,724; R&D), FGF2 (AF-233-NA; R&D), FGF22 (MAB3867; R&D) and IgG (AB-108-C; R&D) were added to the medium.
3
Osteoblast-Megakaryocyte Coculture Protocol
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For coculture experiments, OBs were plated onto 6-well plates (20×103 cells/well). Twenty-four hours later, MKs (20×103 cells/well) were plated into wells for direct coculture. For indirect coculture, OBs were seeded in the upper chamber of a 0.4 μm transwell insert (Corning, New York, USA) and MKs were seeded in the lower wells. Specified cultures were pretreated with 100 nM of SB431542 (Sigma-Aldrich, St. Louis, Missouri, USA) for OBs for approximately 1 h. MKs conditioned medium (MKs-CM) was obtained by removing MKs via centrifugation (5000 rpm, 10 min). In some assays, neutralizing antibodies against TGF-β1 (9016; R&D, Minneapolis, Minnesota, USA ), VEGF (AF-493; R&D), BMP6 (MAB6325; R&D), IGF-1 (AF791; R&D), PDGF-BB (ab34074; Abcam), CXCl12 (79014; R&D), BMP2 (MAB111; R&D), TGF-β2 (AB-112-NA; R&D), TGF-β3 (20724; R&D), BMP4 (MAB50201; R&D) and IgG (AB-108-C; R&D) were added to the MKs-CM.
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