Dasatinib was purchased from LC Laboratories. Roswell Park Memorial Institute (RPMI)‐1640 medium, Dulbecco's modified Eagle's medium (DMEM), foetal bovine serum (FBS), penicillin‐streptomycin and phosphate‐buffered saline (PBS) were bought from WelGENE. 3‐(4,5‐dimethylthiazol‐2‐y)‐5‐(3‐carboxymethoxyphenyl)‐2‐(4‐sulfophenyl)‐2H‐tetrazolium (MTS) reagent was from Promega. Bradford reagent was from Bio‐Rad. Protease inhibitor cocktail (PIC, 100×) and z‐VAD‐fmk (627610) were purchased from Calbiochem. Control siRNA (sc‐37007) and Src (sc‐29228) were obtained from Santa Cruz Biotechnology. Enzyme‐linked chemiluminescence (ECL) Western detection reagents were bought from Thermo Scientific. PCR primers were purchased from Bioneer. LY294002 and PD98059 were from Biomol Research Lab. Cell culture plastic wares were purchased from SPL Life Sciences. A detailed list of antibodies used in this study is included in Supplementary Table S1 .
Control sirna sc 37007
Manufactured by Santa Cruz Biotechnology
Sourced in United States, China, Germany
Control siRNA (sc-37007) is a non-targeting small interfering RNA (siRNA) sequence designed as a negative control for RNA interference (RNAi) experiments. Its core function is to serve as a reference to evaluate the effects of a specific siRNA without confounding off-target effects.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine rnaimax, by Thermo Fisher Scientific (17 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (15 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (7 mentions)
Lipofectamine rnaimax reagent, by Thermo Fisher Scientific (6 mentions)
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Hiperfect transfection reagent, by Qiagen (3 mentions)
Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (3 mentions)
Opti mem medium, by Thermo Fisher Scientific (3 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Dual glo luciferase assay system, by Promega (2 mentions)
Bay11 7082 tlrl b82, by InvivoGen (2 mentions)
Med25 sirna h sc 97106, by Santa Cruz Biotechnology (2 mentions)
Anti rabbit igg na934v, by GE Healthcare (2 mentions)
Peroxidase conjugated anti mouse igg nxa931, by GE Healthcare (2 mentions)
Matrigel 354234, by BD (2 mentions)
Cell lysis buffer 9803, by Cell Signaling Technology (2 mentions)
C myc sirna sc 29226, by Santa Cruz Biotechnology (2 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (2 mentions)
114 protocols using control sirna sc 37007
1
Dasatinib's Anticancer Effects Elucidated
2
Colorectal Cancer Cell Culture
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CCD-18Co (normal colon cells), HT-29 (CRC cells), CaCo2 (CRC cells) and SW480 (CRC cells) cell lines were obtained from the American Type Culture Collection (ATCC; Rockville, MD, USA). The cells were cultured in RPMI-1640 medium (Gibco-BRL, Paisley, UK) supplemented with 10% fetal bovine serum (Hyclone, Logan, UT, USA) at 37°C in 5% CO2. The fibronectin siRNA (sc-29315) and control siRNA (sc-37007) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Transfection was performed using Lipofectamine® 2000 (Invitrogen Life Technologies, Shanghai, China) following the manufacturer's instructions. Negative small interfering RNAs (siRNAs) and scrambled siRNA were synthesized by Shanghai Sangon.
3
Plasmid Transfection and Luciferase Assay
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DNA plasmid constructs were transfected into cells in six-well plates (Corning,), using Lipofectamine 2000 for human cells. Cells were exposed to serum-free medium containing 1 μg DNA of different hiNOS promoter constructs with 10 μg of liposomes for 6 h, washed, and replaced with medium supplemented with 5% calf serum. Cells were lysed with Dual-Glo luciferase Assay System (Promega). Luciferase activity was examined by AutoLumat LB 953 luminometer (Berthold).
Control siRNA (sc-37007) and siRNA against p300 (sc-29431) were obtained from Santa Cruz Biotechnology, and cells were transfected according to the manufacturer's instruction. About 2 × 106 cells were seeded on 6 well cell culture plates one prior to the day of transfection and grew to 60–80% confluent. For each transfection, 50 pmols control siRNA or p300 siRNA were added in siRNA transfection medium. Cells were treated with siRNA medium for 5 hours. The medium were aspirated and replaced with normal growth medium. The sequence of p300 siRNA (sc-29431) is:CCCCUCCUCUUCAGCACCA . The p300 siRNA (sc-29431) was validated in A549 cells by Western Blot (S1 Fig ).
Control siRNA (sc-37007) and siRNA against p300 (sc-29431) were obtained from Santa Cruz Biotechnology, and cells were transfected according to the manufacturer's instruction. About 2 × 106 cells were seeded on 6 well cell culture plates one prior to the day of transfection and grew to 60–80% confluent. For each transfection, 50 pmols control siRNA or p300 siRNA were added in siRNA transfection medium. Cells were treated with siRNA medium for 5 hours. The medium were aspirated and replaced with normal growth medium. The sequence of p300 siRNA (sc-29431) is:
4
Mouse Immune Pathway Silencing Protocol
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The small interfering RNA (siRNA) targeting mouse MDA-5 (sc-61011), RIG-I (sc-51481), IPS-1 (sc-75756), and control siRNA (sc-37007) targeting a scrambled sequence were purchased from Santa Cruz Biotechnology. BAY11-7082 (tlrl-b82) and BX795 (tlrl-bx7) were purchased from InvivoGen (San Diego, CA, USA). In Situ Cell Death Detection Kit (POD 11684817910) was purchased from Roche Diagnostics GmbH (Mannheim, Germany). The antibodies used in this study are listed in Table 1 .
5
Tpr Targeting Protein Detection
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Sources of antibody employed are as follows: anti-His.M8 from Thermo Fisher Scientific was used for western blots; anti-His6 from Roche was used for immunofluorescence microscopy; anti-Tpr from Abcam. Control siRNA (sc-37007) and siRNA targeting Tpr (sc-45343) was purchased from Santa Cruz Biotech.
6
KISS1R Gene Knockdown Protocol
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siKISS1R (sc-60748), and control siRNA (sc-37007) were purchased from Santa Cruz Biotechnology Inc. siKISS1R knockdown was performed using Lipofectamine® RNAiMax transfection reagent (Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions. The efficiency of transfection was determined by mRNA expression.
7
Oxidative Stress Responses in SHSY5Y Cells
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SHSY5Y cells were obtained from ATCC (ATCCCRL-2266) and grown in DMEM/F12 medium (hyclone) supplemented with 10% FBS (EVERY GREEN, Zhejiang Tianhang Biotechnology Co., Ltd, China), 100 μg/mL streptomycin, and 100 U/ml penicillin (Beijing solarbio science & technology co., Ltd) in high humidity condition with 5% CO2 at 37°C. After culturing the cells in 100 mm dishes to reach a ~70% confluence, they were subjected to hydralazine, H2O2 or MPP+ treatment. The dose and duration of application of hydralazine, H2O2 or MPP+ are provided in the figures and text. SiRNA interference were performed by treating the cells with Nrf2 SiRNA (sc-37030) or control SiRNA (sc-37007) (Santa Cruz Biotechnology, Santa Cruz, CA) in 6-well plates for 24 h using the Lipofectamine 3000 reagent (Thermo Fisher Scientific Co., Carlsbad, CA, USA) as indicated in the instructions provided by the manufacturer. After transfection for approximately 24 h, SHSY5Y cells were exposed to hydralazine with or without MPP+. After these treatments, cells were used for biochemical analysis.
8
Silencing GIT1 in HCC Cells
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The specific small interfering RNA (siRNA) sequences against GIT1 (sc-35477) and control siRNA (sc-37007) were both purchased from Santa Cruz Biotechnology. HCC cells were seeded at a concentration of 2×106 per well in six-well plates and cultured overnight. Next, HCC cells were transfected with 100 nM siRNA using Lipofectamine RNA interference MAX Reagent (Invitrogen Life Technologies) according to the manufacturer’s instructions. Then, HCC cells were used for further experiments at 48 hours after transfection.
9
Transient p27 Knockdown via siRNA Transfection
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p27 was downregulated by transiently transfecting siRNA; p27 siRNA (sc-29429) and control siRNA (sc-37007) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, U.S.A.). Cells were plated in 35mm culture plates at 60% confluence in normal growth medium. For each transfection, 100 pmol of siRNA oligomer in 250 μl of serum-free cell medium was mixed with 5 μl Lipofectamine 2000 (diluted 1:50) and incubated for 20 min at room temperature. The complex was then added to the cells and incubated cells at 37°C for 48 h before further experiments.
10
Modulating Hepatic Stellate Cell Activation
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Rat HSC-T6 and human HSC-LX2 cell lines were obtained from the Cell Bank of Chinese Academy of Sciences (Shanghai, China). Rat primary HSCs were isolated as we previously described 15 (link). Rat primary LSECs were purchased from CHI Scientific, Inc. (Wuxi, China). Cells were cultured in Dulbecco's modified eagle medium with 10% fetal bovine serum, 1% antibiotics, and grown in a 5% CO2 humidified atmosphere at 37°C. SMRT siRNA (sc-36514) and NCoR siRNA (sc-36001) for human cells were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Transfection with siRNA against SMRT or NCoR, or control siRNA (sc-37007, Santa Cruz Biotechnology) was performed according to the protocols provided by the manufacturer. Briefly, cells at 60% confluence were transfected with siRNA at a final concentration of 100 nM using the Lipofectamine 2000 Transfection Reagent (Life Technologies, Grand Island, NY, USA) in medium without serum and antibiotics for 24 h. The knockdown efficiency was confirmed by real-time PCR. Recombinant expression plasmids GV230-PPARγ-wild-type and GV230-PPARγ-mutantion were constructed by Shanghai Genechem Co.,Ltd.(Shanghai, China). Their transfection was made by using the Lipofectamine 2000 Transfection Reagent in medium without serum and antibiotics for 24 h.
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