Uterine horns of two adult littermate pairs were collected on 4.5 dpc after mating with a control male. The uterine horns were dissected and placed in ice cold RPMI media containing 1% HEPES, 1% Penicilin-Streptomycin, 1% L-glutamine and 0.5% Bovine Serum Albumin (BSA). Ovaries, oviduct and the cervix were removed and the remaining uterine horn was mechanically dissociated using blades. The minced tissue was placed in dissociation mix of the abovementioned media with collagenase type II, Dispase and DNAse I (Sigma) and incubated for 40 minutes at 37°C. After incubation, 0.5 mM EDTA was added and the tissue was further dissociated using a glass pipette. The remainder of the tissue was incubated for an additional 5 minutes at 37°C. The suspension was passed through a 70 μm filter and the flow through was centrifuged at 1,200 rpm for 5 minutes in an Eppendorf centrifuge (model 5702f). The cell pellet was resuspended and stained with Alexa Fluor 647 anti-mouse CD31 Antibody (# 102516, Biolegend), Alexa Fluor® 488 anti-mouse CD45 antibody (#103121, Biolegend) and DAPI for 20 minutes on ice. Endothelial cells (Cd31+, Cd45−, DAPI−) were sorted using a FACSJazz (BD Biosciences, San Jose, CA) cell sorter and RNA was immediately extracted using an Arcturus PicoPure RNA Isolation Kit (Applied Biosystems) according to the manufacturer’s instructions.
Alexa fluor 647 anti mouse cd31 antibody
Manufactured by BioLegend
Sourced in United States
Alexa Fluor 647 anti-mouse CD31 Antibody is a fluorescent-labeled antibody that binds to the CD31 cell surface antigen expressed on mouse endothelial cells. It can be used for the identification and characterization of mouse endothelial cells in flow cytometry and other immunoassays.
Automatically generated - may contain errors
Lab products found in correlation
Dnase 1, by Merck Group (1 mentions)
Facsjazz, by BD (1 mentions)
Dispase, by Merck Group (1 mentions)
Alexa fluor 488 anti mouse cd45 antibody, by BioLegend (1 mentions)
Collagenase type 2, by Merck Group (1 mentions)
Vettest 8008, by IDEXX (1 mentions)
C57bl 6 mice, by Jackson ImmunoResearch (1 mentions)
Caspase glo 3 7 assay kit, by Promega (1 mentions)
Coumarin 6, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Polyvinyl alcohol (pva), by Merck Group (1 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (1 mentions)
3 protocols using alexa fluor 647 anti mouse cd31 antibody
1
Endothelial Cell Isolation from Uterine Horn
2
LPS-Induced Acute Kidney Injury
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CD11c-YFP (Itgax-Venus) mice and C57BL/6 mice were purchased form The Jackson Laboratory (USA) and Junbiotech (Gyeonggi, Korea). CD11c-YFP mice were used for imaging experiments, while C57BL/6 mice were used for histological analysis and immunofluorescence staining for F4/80. To induce acute kidney injury, CD11c-YFP mice were intraperitoneally injected with LPS (5 µg/g), whereas the control group (n=7) was injected with the same volume of 0.9% saline. The LPS-treated mice were pretreated with either MCC950 (10µg/g, intraperitoneally, n=9) or 0.9% saline (n=7) 1 hour before LPS administration. Mice were euthanized, and kidneys were harvested 24 hours after LPS treatment. Mice received 10 μg of Alexa Fluor ® 647 anti-mouse CD31 antibody (BioLegend®, 102516, 0.5 μg/μl) via the tail vein 10 min before euthanasia. Blood urea nitrogen and serum creatinine levels were determined using the VetTest 8008 (IDEXX Laboratories, Westbrook, ME, USA). All animal experiments were performed in compliance with the guidelines of the Animal Research Ethics Committee of Kyung Hee University Hospital at Gangdong (KHNMC AP 2019-014).
3
Targeted Nano-formulation for Cancer Therapy
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Monomethoxy-poly(ethylene glycol)-poly lactic-co-glycolic acid (PEG-PLGA) was obtained from Jinan Dai Gang Biomaterial Co., Ltd. (Jinan, China). PTX was purchased from Dalian Melone Biomart Co., Ltd, and iRGD (CRGDRGPDC) was provided by Genscript Co., Ltd. (Guilin, China). Coumarin-6 and polyvinylalcohol (PVA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cell counting kit-8 (CCK-8), Annexin V apoptosis detection kits, and PI/RNase cell cycle assay kits were obtained from Dojindo (Kumamoto, Japan). Caspase-Glo® 3/7 assay kits were purchased from Promega (Madison, WI, USA), and 4,6-diamidino-2-phenylindole (DAPI) was obtained from Molecular Probes (Eugene, OR, USA). Alexa Fluor 647 anti-mouse CD31 antibody was purchased from Biolegend (San Diego, CA, USA), and 1,1ʹ-dioctadecyl-3,3,3ʹ,3ʹ- tetramethylindodicarbocyanine (DID) was provided by Key GEN Bio TECH Co., Ltd. Anti-integrin and neuropilin-1 antibodies were purchased from Abcam (Cambridge, MA, USA).
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