Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), trypsin/EDTA, and penicillin/streptomycin were from Gibco (Life Technologies, Grand Island, NY, USA). Antibodies of glyceroaldehyde-3-dehydrogenase (GAPDH) were purchased from Santa Cruz, whereas antibodies for Ac-H3, OPG and RANKL were from GeneTex. Ethidium bromide, agarose and kits for reverse transcription (RT) and polymerase chain reaction (PCR) were purchased from HT Inc., UK. Total RNA isolation kits were from Qiagen Inc. (Santa Clarita, CA, USA). Specific PCR primer sets were synthesized by Genemed Biotechnologies, Inc. (San Francisco, CA, USA). Protein assay kits were obtained from Bio-Rad (Bio-Rad Labs, Hercules, CA, USA). Sodium butyrate was obtained from Sigma (Sigma-Aldrich Company, St. Louis, MO, USA). MG-63 cells were obtained from the American Type Culture Collection (ATCC). OPG, RANKL, pro-collagen I, MMP-2, osteopontin, osteocalcin and osteonectin (SPARC) ELISA kits were from R&D. 8-Isoprostane ELISA kits were from Cayman.
Total rna isolation kit
Manufactured by Qiagen
Sourced in United States, Germany
The Total RNA Isolation Kit is a product designed for the extraction and purification of total RNA from a variety of sample types. It utilizes a silica-based membrane technology to efficiently capture and recover RNA molecules, including both small and large RNA species. The kit provides a standardized protocol and the necessary reagents to enable reliable and consistent RNA isolation.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (3 mentions)
M mlv reverse transcriptase, by Promega (3 mentions)
Oligo dt 15 primer, by Promega (3 mentions)
Cfx connect, by Bio-Rad (2 mentions)
Superscript 3 first strand cdna synthesis kit, by Thermo Fisher Scientific (2 mentions)
Ssoadvanced sybr green supermix, by Bio-Rad (2 mentions)
Cfx96 real time pcr detection system, by Bio-Rad (2 mentions)
Protein assay kit, by Bio-Rad (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Sodium butyrate, by Merck Group (1 mentions)
Mmp 2, by R&D Systems (1 mentions)
Mg 63 cells, by American Type Culture Collection (1 mentions)
Pro collagen 1, by R&D Systems (1 mentions)
Rankl, by R&D Systems (1 mentions)
Osteopontin, by R&D Systems (1 mentions)
Fatty acid, by Merck Group (1 mentions)
Retinol, by Merck Group (1 mentions)
C peptide, by Merck Group (1 mentions)
β carotene, by Merck Group (1 mentions)
30 protocols using total rna isolation kit
1
Osteoblast Differentiation Protocol
2
Comprehensive Analytical Methods for Metabolic Assessment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All chemicals used were of analytical grade. Triglycerides and glucose assay kits were purchased from BioSystems SA (Barcelona, Spain). Alanine aminotransferase (ALT), aspartate aminotransferase (AST), and retinol-binding protein 4 (RBP4) measurement kits were procured from BioVision Inc. (Milpitas, CA, USA). Recombinant human insulin, β-hydroxy butyrate assay kit, secondary antibodies, various standards such as retinol, β-carotene, β-apo-8′-carotenal, and fatty acids were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). C-peptide (EMD Millipore Corporation, Billerica, MA, USA), insulin (Crystal Chem Inc., Downers Grove, IL, USA), and fibroblast growth factor 21 (FGF21) (R&D Systems, Minneapolis, MN, USA) quantitation kits were used. Total RNA isolation kits were obtained from Qiagen GmbH (Hilden, Germany). For quantitative real-time polymerase chain reaction analysis (qRT-PCR), first-strand cDNA synthesis kits (New England Biolabs, Ipswich, MA, USA) and pre-validated universal probe for rats (Roche Diagnostics GmbH, Mannheim, Germany) were used. Experimental diets were obtained from OpenSource Diets (Research Diets Inc., New Brunswick, NJ, USA).
3
Inhibitors of Cell Signaling Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell culture medium and related reagents were obtained from Life Technologies (Thermo Fisher Scientific Ltd., Waltham, MA, USA). Dimethyl sulfoxide (DMSO), LY294002 (a PI3K/Akt inhibitor), U0126 (1,4-diamino-2,3-dicyano-1,4-bis (2-aminophenylthio)butadiene) (a MEK/ERK inhibitor), and SB203580 (a p38 inhibitor), were purchased from Sigma–Aldrich (St. Louis, MO, USA). 5Z-7-Oxozeaenol (a TAK1 inhibitor) was purchased from Tocris Cookson Ltd. (Bristol, UK). Total RNA isolation kits were obtained from Qiagen (Taipei, Taiwan). Polymerase chain reaction (PCR) primers for β–actin (BAC), uPA, uPAR, and PAI-1 were synthesized by Genemed Biotechnologies, Inc. (San Francisco, CA, USA). Enzyme-linked immunosorbent assay (ELISA) kits for uPA and soluble uPAR (suPAR) were obtained from R&D Systems (R&D DuoSet, Minneapolis, MN, USA). Recombinant IL-1β and PAI-1 ELISA kits were obtained from PeproTech (PeproTech Asia, Rehovot, Israel).
4
Transcriptomic Analysis of P. argentipes Resistance
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was isolated from laboratory reared single P. argentipes DDT exposed and non-exposed resistant and susceptible P. argentipes by using Qiagen total RNA isolation Kit. The cDNA was synthesized from 1.0 µg of isolated RNA using RevertAid First Strand cDNA Synthesis kit (Thermo Fisher) through random hexamer primers. Primers for real-time PCR were designed using the Parg-GSTσ gene sequence deduced from this study. The primers used were forward 5′-CACTGAGAAATATCCCAACCTCA-3′ and reverse 5′-TTAAACTTCAGTAACAGGTCGTTTC-3′. Real-Time PCR reaction was performed on a Qiagen multiplex RT-PCR and the relative expression and fold change was calculated according to the 2-ΔΔCT method, incorporating PCR efficiency49 (link) after normalization with the housekeeping gene 28 s rRNA.
5
Total RNA Isolation and Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA was extracted using a total RNA isolation kit (Qiagen, Hilden, Germany) according to the manufacturer’s protocol. RNA concentrations were measured using a spectrophotometer (ND-1000, NanoDrop Technologies, Wilmington, DE, UnitedStates). Samples with a 260/280 ratio of ∼2.0 were used for further processing.
6
Quantitative RT-PCR for Gene Expression
7
Isolation and Analysis of NK Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NK cells were sorted from spleens on days 1, 3, and 7 after MCAO or the sham operation as previously described [3 (link)]. TRIzol Reagent (Thermo Fisher Scientific, Waltham, MA, USA) and a total RNA isolation kit (Qiagen, Germantown, MD, USA) were used according to the manufacturer’s protocol to isolate total RNA from NK cells pooled from 15 mice per data point. Then, the RNAs were analyzed with an nCounter miRNA Expression Assay (NanoString Technologies, Inc., Seattle, WA, USA), which can detect multiple miRNAs.
8
Quantifying Endothelial Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total cellular RNA was extracted from HUVECs using a Total RNA Isolation Kit (Qiagen) according to the manufacturer’s instructions. Complementary DNA (cDNA) was synthesized using M-MLV reverse transcriptase (Promega) and oligo-dT 15 primer (Promega). cDNA was amplified by PCR over 30 cycles. The following oligonucleotide primers were used in this study: Human eNOS: sense 5′-TGATGCATTGGATCTTTGGA-3′ and antisense 5′-CCATGTTACTGTGCGTCCAC-3′; Human KLF2: sense 5′-CCTCCCAAACTGTGACTGGT-3′ and antisense 5′-ACTCGTCAAGGAGGATCGTG-3′; Human VCAM-1: sense 5′-CCGGATTGCTGCTCAGATTGGA-3′ and antisense 5′-AGCG TGGAATTGGTCCCCTCA-3′; Human ICAM-1: sense 5′-GG CCTCAGTCAGTGTGA-3′ and antisense 5′-AACCCCATTCAG CGTCA-3′; Human GAPDH: sense 5′-GAGTCAACGGATTTG GTCGT-3′ and antisense 5′-TTGATTTTGGAGGGATCTCG-3′.
9
Transcriptome Analysis of Asthmatic Airway Smooth Muscle Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human ASMC were obtained from bronchial biopsies and explanted lungs from doctor diagnosed asthmatic patients (n=3) and healthy controls (n=3). ASMC were isolated and grown in culture as previously described.10 (link)
Total cellular mRNA was isolated using the Qiagen total RNA isolation kit (Qiagen, Doncaster, Victoria, Australia). Samples were labelled and run on an Affymetrix (Santa Clara, California, USA) GeneChip Human Gene 1.0 ST Arrays according to the manufacturer's instructions (GSE63383). Microarray analysis was conducted using R software V.3.02, using the Bioconductor-limma package, and normalised using Robust Multi-array Average.
Total cellular mRNA was isolated using the Qiagen total RNA isolation kit (Qiagen, Doncaster, Victoria, Australia). Samples were labelled and run on an Affymetrix (Santa Clara, California, USA) GeneChip Human Gene 1.0 ST Arrays according to the manufacturer's instructions (GSE63383). Microarray analysis was conducted using R software V.3.02, using the Bioconductor-limma package, and normalised using Robust Multi-array Average.
10
Comprehensive Adipokine Profiling in Rats
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Triglyceride assay kit was purchased from BioSystems (BioSystems S.A., Barcelona, Spain). Free fatty acid (FFA) (Biovision Inc., CA, USA), leptin, tumor necrosis factor α (TNFα) macrophage chemoattractant protein 1 (MCP1) (Life Technologies, Thermo Fischer Scientific Inc., MD, USA), high sensitive C-reactive protein (hsCRP) (CUSABIO, Hubei, China) and adiponectin (Crystal Chem Inc., IL, USA) assays kits were procured. Testosterone and estradiol (E2/17β-estradiol) assay kits were from Calbiotech (Calbiotech Inc. CA, USA). Proteome Profiler Rat Adipokine Array kit was from R&D Systems, MN, USA. Total RNA isolation kit was obtained from Qiagen (Qiagen GmbH, Hilden, Germany). For quantitative real-time PCR analysis (qRT-PCR), First strand cDNA synthesis kit (New England Biolabs, Ipswich, MA, USA) and pre-validated universal probe for rat (Roche diagnostics GmbH, Mannheim, Germany) were used. Experimental diets were obtained from OpenSource Diets (Research Diets Inc., New Brunswick, NJ, USA).
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!