All the overexpression plasmids targeting VHL (pcDNA3.1-VHL), HIF-1α (pcDNA3.1-HIF-1α), and ZEB1 (pcDNA3.1-ZEB1) were designed and synthesized by GeneChem (China), and the empty plasmid was used as a negative control. HIF-1α siRNA (siHIF-1α), VHL siRNA (siVHL), ELK1 siRNA (siELK1), ZEB1 siRNA (siZEB1), and corresponding negative control were purchased from RiboBio (Guangzhou, China). MiR-21-5p mimics, miR-21-5p inhibitors, and matched control (mimic-NC or inhibitor-NC) were synthesized by Invitrogen (Shanghai, China). All the small interference RNAs were transfected at a final concentration of 50 nM, and the plasmid was transfected at a final concentration of 0.2 μg for 96 well plates and 1.6 μg for 12 well plates. Lipofectamine 3000 (Invitrogen, USA) was used for cell transfections according to the manufacturer’s instructions. Protein and total RNA were extracted after 48 h.
Inhibitor nc
Manufactured by Thermo Fisher Scientific
Sourced in United States
Inhibitor-NC is a laboratory product designed to inhibit or block the activity of certain biological molecules or processes. It is a tool used in research and scientific applications. The core function of Inhibitor-NC is to provide a way to study the effects of inhibiting specific targets, without making claims about its intended use or applications.
Automatically generated - may contain errors
Lab products found in correlation
Nc mimic, by Thermo Fisher Scientific (21 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (15 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (10 mentions)
Inhibitor nc, by GenePharma (10 mentions)
Mimic nc, by GenePharma (8 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (8 mentions)
Mimics nc, by Thermo Fisher Scientific (5 mentions)
Pcdna3.1 vector, by Thermo Fisher Scientific (4 mentions)
Inhibitor nc, by RiboBio (4 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (4 mentions)
Mir nc, by Thermo Fisher Scientific (3 mentions)
Sh nc, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Nc sirna, by Thermo Fisher Scientific (3 mentions)
Mir 21 5p mimic, by Thermo Fisher Scientific (2 mentions)
Polybrene, by Merck Group (2 mentions)
Mir nc, by RiboBio (2 mentions)
Mimic nc, by RiboBio (2 mentions)
Inhibitor nc, by Genechem (2 mentions)
Pcdna3, by GenePharma (2 mentions)
44 protocols using inhibitor nc
1
Overexpression and Knockdown of Key Regulators
2
NEAT1 Silencing and BCL2L11 Overexpression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Synthesized NEAT1 small interference RNA (si-NEAT1) was cloned into the vector (pAdTrack-CMV, GenePharma, Shanghai, China) and scrambled siRNA (si-nc) was cloned into the vector as a negative control. HEK-293 cells were used to pack lentivirus by transfection with vectors containing si-NEAT1 or si-nc. After 72 h of transfection, the cells were infected with virus particles using Polybrene (Sigma, St Louis, MO).
To obtain BCL2L11 overexpression plasmid, BCL2L11 was first amplified from cDNA using PCR and then inverted into pcDNA3.1 vector (Invitrogen, Carlsbad, CA) with the name of pcDNA3.1-BCL2L11.
miR-29a inhibitor, miR-29a mimics, inhibitor nc, and mimics nc were purchased from GenePharma (Shanghai, China).
After vector construction, the treated cells were transfected with si-nc, si-NEAT1, miR-29a inhibitor, inhibitor nc, miR-29a mimics, mimics nc, pcDNA3.1, or pcDNA3.1-BCL2L11 using Lipofectamine 2000 (Invitrogen, Carlsbad, CA).
To obtain BCL2L11 overexpression plasmid, BCL2L11 was first amplified from cDNA using PCR and then inverted into pcDNA3.1 vector (Invitrogen, Carlsbad, CA) with the name of pcDNA3.1-BCL2L11.
miR-29a inhibitor, miR-29a mimics, inhibitor nc, and mimics nc were purchased from GenePharma (Shanghai, China).
After vector construction, the treated cells were transfected with si-nc, si-NEAT1, miR-29a inhibitor, inhibitor nc, miR-29a mimics, mimics nc, pcDNA3.1, or pcDNA3.1-BCL2L11 using Lipofectamine 2000 (Invitrogen, Carlsbad, CA).
3
Molecular Regulation of Cardiac Function
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MiR-33a-5p mimics (mimics-miR-33a-5p), miR-33a-5p inhibitor, and their negative controls (miR-NC and inhibitor-NC, respectively) were designed and synthesized by RiboBio (Guangzhou, China). Small interfering RNAs (siRNAs) targeting USP46 and circ-NNT (si-USP46 and si-circ-NNT, respectively) and a scrambled form used as control (shRNA NC) were obtained from Dharmacon (Lafayette, CO, USA). Mimics-miR-33a-5p, mimics-NC, miR-33a-5p inhibitor, inhibitor-NC, si-USP46, circ-NNT shRNA, and shRNA NC were transfected into cultured cardiomyocytes using Lipofectamine® 2000 (Invitrogen) following manufacturer’s instructions. The circ-NNT exon along with the endogenous flanking sequence (1 kb upstream) was inserted into the pcDNA3.1 vector, and part of the upstream flanking sequence was inserted in an inverted orientation downstream. The mouse USP46 was cloned by PCR using mouse cDNA as the template and inserted into the pcDNA3.1 vector. All vectors (pcDNA-circ-NNT, pcDNA-USP46, and pcDNA3.1 empty vector) as well as si-USP46, circ-NNT shRNA, and shRNA NC were cloned into the Adeno-X Expression System (Clontech, Otsu, Japan) following manufacturer’s instructions. All adenoviral constructs were amplified in HEK293 cells. Adenoviral infection of HEK293 cells or cardiomyocytes was carried out as previously described [39 (link)].
4
Modulation of miR-146a-5p in Caco-2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
miR-146a-5p mimics, inhibitors, mimic negative control (NC) and inhibitor NC were obtained from Sangon Biotech, Co., Ltd. The sequences of miR mimic, inhibitor and NC were as follows: miR-146a-5p mimics, 5'-UGAGAACUGAAUUCCAUGGGUU-3'; miR-146a-5p inhibitors, 5'-AACCCAUGGAAUUCAGUUCUCA-3'; miRNA mimic NC, 5'-UUGUACUACACAAAAGUACUG-3' and miRNA inhibitor NC, 5'-CAGUACUUUUGUGUAGUACAA-3'. A total of 500,000 Caco-2 cells were plated in 48-well plates with 200 µl/well DMEM supplemented with 1,000 mg/l glucose and 20% FBS. One day before transfection. Cells (50-70% confluence) were transfected with 10 pmol miR-146a-5p mimics, inhibitors, mimic NC or inhibitor NC using 0.5 µl Lipofectamine® 2000 (Invitrogen; cat. no. 11668-019; Thermo Fisher Scientific, Inc.) resuspended in 100 µl DMEM (Gibco; cat. no. C11995500BT). A volume of 200 µl transfection suspension was added to each well and incubated at 37˚C for 5 h. Then we replaced the transfection suspension with 20% complete medium and continued culture for 24 h before subsequent experiments.
5
Investigating miR-373 Regulation of GAB2 in Lung Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
miR-373 mimics, mimics negative control (NC), inhibitor and inhibitor NC were synthesized by Genepharma (Shanghai, China). The sequences are as follows: MiR-373 mimics, 5′-GAAGUGCUUCGAUUUUGGGGUGU-3′; mimics NC, 5′-GCUUUAUUAGAGUGCUAUUGCUU-3′; miR-373 inhibitor, 5′-ACACCCCAAAAUCGAAGCACUUC-3′; and inhibitor NC, 5′-AUUCUCAUCACCAAUCACGAAAUA-3′. A549 and H358 cells were transfected using Lipofectamine® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) in accordance with the manufacturer's instructions with a GAB2-overexpression pcDNA3.1 plasmid (1 µg; Invitrogen; Thermo Fisher Scientific, Inc.). As an NC, a pcDNA3.1 vector that was empty was utilized. After 48 h of transfection, the cells were harvested for the following experiments. The efficiency of overexpression or inhibition was verified by RT-qPCR and western blot.
6
Modulating miR-221-3p in DEF Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
miR-221-3p mimic, mimic negative control (mimic-NC), miR-221-3p inhibitor, and inhibitor negative control (inhibitor-NC) were purchased from RiboBio (Guangzhou, China). DEF cells were seeded into 6-well or 12-well plates and cultured in DMEM with 10% FBS at 37°C. When the DEF cells were grown to 70–80% confluence, the miRNA mimic (100 nmol), mimic-NC (100 nmol), inhibitor (200 nmol), and inhibitor-NC (200 nmol) were transfected into the cells using Lipofectamine 3000 (Invitrogen, United States) according to the manufacturer’s protocol. After 24 h of transfection, the DEF cells were infected with the DTMUV CQW strain. Then, the cells were collected for future analysis at 36 or 72 h after infection.
7
LINC00662 Modulation in Osteosarcoma Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Small hairpin (sh)-LINC00662, the negative control vector (sh-NC), miR-NC, miR-30b-3p mimics, inhibitor NC, miR-30b-3p inhibitor, pcDNA-NC, and pcDNA-ELK1 were purchased from RiboBio Company (Beijing, China). When the cultured cells reached 80–90% confluence, the above factors were transfected into U2OS and MG63 cells using Lipofectamine 3000 Transfection Reagent (Invitrogen) for 48 h. The sequences of transfected fragment were as follows: miR-NC, 5’-CAGUACUUUUGUGUAGUACAA-3’; miR-30b-3p mimics, 5’-UGUAAACAUCCUACACUCAGCU-3’; inhibitor NC, 5’-UCACAACCUCCUAGAAAGAGUAGA-3’; miR-30b-3p inhibitor, 5’-AGCUGAGUGUAGGAUGUUUACA-3’; sh-LINC00662, 5’-GCUGCUGCCACUGUAAUAAUU-3’; sh-NC, 5’-AAUUCUCGGAACGUCUGACGU-3’.
8
Modulating miR-103a-3p and FBXW7 in Cervical Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
miR-103a-3p mimics (5′-AGCAGCATTGTACAGGGCTATGA-3′), mimics negative control (NC) (5′-ATAGTGATCAGATGGGCAGCCTA-3′), miR-103a-3p inhibitor (5′-TCATAGCCCTGTACAATGCTGCT-3′), inhibitor NC (5′-TATCCCACGCTAGCTGTCTTGAA-3′) were obtained from Shanghai GenePharma Co., Ltd. pcDNA3.1-FBXW7, pcDNA3.1 vector, si-FBXW7 (5′-GCTCCCTAAAGAGTTGGCACTCTAT-3′) or si-Scramble (5′-GCTATCGAATGAGGTCACCTCCTAT-3′) were synthesized by Guangzhou RiboBio Co, Ltd. (Guangzhou, China). Then, SiHa and Hela cells (4 × 105 cells per well) were seeded in 6-well plates and cultured overnight until reached about 80% confluence, 100 nM miR-103a-3p mimics, mimics NC, miR-103a-3p inhibitor, inhibitor NC, 2 μg pcDNA3.1-FBXW7 plasmid, and 100 ng si-FBXW7 were transfection using the Lipofectamine 3000 reagent (Invitrogen, Thermo Fisher Scientific, Inc., Waltham, MA, USA) according to the manufacturer’s protocol. After incubation for 6 h, regular culture medium was added into each well to terminate reaction at 37 °C. 48 post-transfection, cells were harvested for subsequent experiments.
9
Regulation of MALAT1, miR-206, ARNT, and PPARα in HepG2 cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The shRNA MALAT1 (sh-MALAT1), sh-negative control (sh-NC), miR-206 inhibitor, inhibitor NC, miR-206 mimics, miR-NC, sh-ARNT and sh-PPARα were obtained from GenePharma (Shanghai, China). HepG2 cells transfected with sh-MALAT1, sh-NC, miR-206 inhibitor, inhibitor NC, miR-206 mimics, miR-NC, sh-ARNT or sh-PPARα by using Lipofectamine 3000 (Invitrogen).
10
Overexpression and Silencing of lncRNA ANRIL
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Full-length human lncRNA ANRIL sequences were amplified and ligated into pcDNA3.1 plasmid (Invitrogen, Carlsbad, CA, USA), and the recombined plasmid was referred to as pc-ANRIL. The stable knockdown lncRNA ANRIL vector (si-ANRIL) was generated by the specific target small interference RNA (siRNA). Empty pcDNA3.1 or pc-ANRIL, as well as si-ANRIL were transfected into HLEC SRA01/04 cells with the help of the lipofectamine 3000 reagent (Invitrogen) according to the manufacturer’s instructions. Stable cell line overexpressing lncRNA ANRIL was obtained after selection in culture medium containing 0.5 mg/mL G418 (Sigma-Aldrich) for approximately 4 weeks. MicroRNA (miR)-21 inhibitor, miR-34a inhibitor and miR-122-5p inhibitor as well as its negative control (inhibitor-NC) were synthesized by Shanghai GenePharma Co., Ltd. (Shanghai, China). The lipofectamine 3000 reagent (Invitrogen) was also used for transfection with miR-21 inhibitor or inhibitor-NC. Cells were harvested at 72 h post-transfection in the subsequent experiments.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!