miR-9-5p mimic control (miR-Con), miR-9-5p mimic (miR-9), miR-9-5p inhibitor control (anti-miR-Con), and miR-9-5p inhibitor (anti-miR-9) were synthesized by Ribobio (Guangzhou, China). Primary mouse microglia and BV-2 cells were seeded at 1 × 105 cells per well in a 24-well plate in 1 ml of antibiotic-free standard growth medium and grown to 40–80% confluency. Then, cells were transfected with 50 nM miR-Con or miR-9 or with 100 nM anti-miR-Con or anti-miR-9. 1 μl RiboFECT™CP reagent (RiboBio, Guangzhou, China) was added to each well according to the manufacturer’s protocols. The medium containing RiboFECT™CP reagent was replaced after 24 h of transfection. Transfection efficiency was determined via western blotting.
Anti mir con
Manufactured by RiboBio
Sourced in China
Anti-miR-con is a laboratory reagent designed to serve as a control for experiments involving anti-miRNA (microRNA) inhibition. It is used to help evaluate the specificity and efficacy of anti-miRNA treatments in various experimental settings.
Automatically generated - may contain errors
Lab products found in correlation
Mir con, by RiboBio (6 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (2 mentions)
Si con, by RiboBio (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Mir 9 5p mimic, by RiboBio (1 mentions)
Anti mir 9, by RiboBio (1 mentions)
Ribofect cp reagent, by RiboBio (1 mentions)
Mir 495 mimic, by RiboBio (1 mentions)
Pcdna3.1 circrna mini vector, by Addgene (1 mentions)
Preceiver m02 erbb3, by GeneCopoeia (1 mentions)
Pcdna3.1 empty vector, by Thermo Fisher Scientific (1 mentions)
Si neat1, by RiboBio (1 mentions)
Mir 214 3p, by RiboBio (1 mentions)
Dual lucy assay kit, by Vigorous Biotechnology (1 mentions)
Pcdna empty vector pcdna, by RiboBio (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
8 protocols using anti mir con
1
Transfection of miR-9-5p in Mouse Microglia
2
Modulation of circSERPINE2 in CHON-001 Cells
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CircSERPINE2 overexpression vector was synthesized via cloning circSERPINE2 (hsa_circ_0008365) sequence into pcDNA3.1 circRNA mini vector, with the pcDNA3.1 circRNA mini vector (Addgene, Cambridge, MA, U.S.A.) as negative control (pcDNA). siRNA for circSERPINE2 (si-circSERPINE2, 5′-UCCGGUGUGGUCGUCCUUGGU-3′), negative control of siRNA (si-con, 5′-AAGACAUUGUGUGUCCGCCTT-3′), miR-495 mimic (5′-AAACAAACAUGGUGCACUUCUU-3′), negative control of mimic (miR-con, 5′-ACGUGACACGUUCGGAGAATT-3′), miR-495 inhibitor (anti-miR-495, 5′-AAGAAGUGCACCAUGUUUGUUU-3′), and negative control of inhibitors (anti-miR-con, 5′-CAGUACUUUUGUGUAGUACAA-3′) were synthesized via Ribobio (Guangzhou, China). For cell transfection, CHON-001 cells with 60% confluence were transfected with the vectors or oligonucleotides (30 nM) using Lipofectamine 2000 (Thermo Fisher) for 24 h.
3
Regulation of Smad3 by miR-129-3p in H9c2 cells
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Pre-miR-Con, pre-miR-129-3p, anti-miR-Con and anti-miR-129-3p were synthesized by RiboBio (Guangzhou, China). H9c2 cells were seeded in 6-well plates and transfected with pre-miR-Con, pre-miR-129-3p, anti-miR-Con and anti-miR-129-3p using Lipofectamine 2000 (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA) for 48 h at 37 °C according to the manufacturer’s protocol.
A mammalian expression plasmid (pReceiver-M02-ERBB3; GeneCopoeia, Germantown, MD, USA) was designed to specially express the full-length open reading frame (ORF) of rat Smad3 without miR-129-3p responsive 3′-UTR. An empty plasmid served as a negative control. Overexpressed Smad3 plasmids (vector-AGTRAP) and control plasmids (vector-Con) were transfected into H9c2 cells using Lipofectamine 2000 (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA) for 48 h at 37 °C according to the manufacturer’s protocol.
A mammalian expression plasmid (pReceiver-M02-ERBB3; GeneCopoeia, Germantown, MD, USA) was designed to specially express the full-length open reading frame (ORF) of rat Smad3 without miR-129-3p responsive 3′-UTR. An empty plasmid served as a negative control. Overexpressed Smad3 plasmids (vector-AGTRAP) and control plasmids (vector-Con) were transfected into H9c2 cells using Lipofectamine 2000 (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA) for 48 h at 37 °C according to the manufacturer’s protocol.
4
Lipofectamine-mediated Transfection of miRNA, siRNA, and Plasmids
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Cells were transfected with siRNAs, miRNAs, or plasmids using Lipofectamine 2000 (Life Technologies, Carlsbad, CA) according to the manufacturer's instructions. miRNA mimics, the miR-215 inhibitor (anti-miR-215), and the respective negative controls (miR-Con and anti-miR-Con) were purchased from RiboBio (Guangzhou, China). Control and Dicer siRNAs were obtained from Life Technologies and the sequences were described previously 16 (link). The human Dicer overexpression plasmid pDESTmycDICER (pDicer) was obtained from Addgene (Cambridge, MA).
5
KCNQ10T1 Regulation by miR-329-3p and CTNND1
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Small interfering RNAs specific to KCNQ10T1 (termed si-KCNQ10T1#1/2) and negative control siRNA (si-con) were synthesized and purchased from RiboBio Company (Guangzhou, China). MiR-329-3p mimic, miR-con, miR-329-3p inhibitor (anti-miR-329-3p), and anti-miR-con were obtained from RiboBio (Guangzhou, China). Moreover, to over-express CTNND1, the sequence of CTNND1 was cloned into a pcDNA3.1 empty vector (Invitrogen), named as pcDNA-CTNND1, and empty vector was considered as a negative control (pcDNA). According to the manufacturer’s instructions, all these oligonucleotides and plasmids were transfected into SW480 and LS1034 cells by using Lipofectamine 3000 reagent (Invitrogen).
6
Modulating MIR22HG in Cardiomyocytes
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MIR22HG-overexpressing plasmid pcDNA-MIR22HG (termed MIR22HG) and its corresponding control (termed Vector), small interfering RNAs targeting MIR22HG (termed si-MIR22HG-1 and si-MIR22HG-2) and their scramble control (termed Scramble), and miR-24 inhibitor (termed anti-miR-24) and inhibitor control (termed anti-miR-con) were designed and synthesized by RiboBio Co., Ltd (Guangzhou, China). Transient transfection of AC16 cardiomyocytes with oligonucleotides or plasmids was carried out using Lipofectamine 3000 reagent (Invitrogen, Carlsbad, CA, USA).
7
siRNA-mediated NEAT1 regulation
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The specific siRNAs of NEAT1 and scrambled control (si-NEAT1, si-con) were synthesized by RiboBio Corporation (Guangzhou, People’s Republic of China). The ectopic vector pcDNA3.1-NEAT1 (pc-NEAT1) and its control (pc-con) were constructed by Thermo Fisher Scientific. The miR-214-3p mimics/inhibitors with corresponding controls (miR-214-3p, anti-miR-214-3p, miR-con and anti-miR-con) were purchased from RiboBio. The Wnt signaling quantitation luciferase reporter plasmids (TOP Flash, FOP Flash) were purchased from BioVector NTCC Inc. (Beijing, People’s Republic of China). Transient transfection was carried out using Lipofectamine™ 3,000 (Thermo Fisher Scientific) following the manufacturer’s instructions. The reporter activities were determined 48 hours post-transfection by the Dual-Lucy Assay Kit from Vigorous Biotechnology (Beijing, People’s Republic of China), with firefly luciferase as base line and renilla luciferase as internal control described as before.16 (link)
8
Ox-LDL-induced HAEC Dysfunction
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Primary human aortic endothelial cells (HAECs) were obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured in RPMI-1640 medium (Gibco, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; GE Healthcare Life Sciences, Logan, UT, USA), 100 U/mL penicillin G, and 100 µg/mL streptomycin at 37℃ in a 5% CO2 atmosphere. miR-590 mimics (miR-590), miRNA negative control (miR-con), miR-590 inhibitors (anti-miR-590), inhibitor negative control (anti-miR-con), siRNA specially against TLR4 (si-TLR4), siRNA negative control (si-con), pcDNA-TLR4 (TLR4), and pcDNA empty vector (pcDNA) were obtained from RiboBio Co., Ltd. (Guangdong, China). When grown to 70–80% confluence, HAECs were transiently transfected with miRNAs (100 nM), siRNAs (100 nM), and pcDNA plasmids (1 µg/mL) using Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA). One day after transfection, HAECs were exposed to 100 µg/mL of ox-LDL (UnionBiol, Beijing, China) for a further 24 h.
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