Sodium new houttuyfonate (SNH) was purchased from Xi'an Kailai Biological Engineering Co., Ltd. (Xi'an, China). We used high performance liquid chromatography to quantitatively analyze SNH with a purity of 98% (Fig. S1† ). HepG2 cells were obtained from ATCC. Dulbecco's Modified Eagle Medium (DMEM) was purchased from Gibco (USA). Fetal bovine serum (FBS), MTT, penicillin and streptomycin all were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Annexin V-FITC/PI apoptosis detection kit, Apoptosis-Hoechst staining kit and DNA ladder extraction kit were from Shanghai Beyotime Biological Technology Co. Ltd. (Shanghai China). The primer of GADPH, MMP9, VEGF and Bcl-2 were synthesized by Shanghai Sangon Biological Engineering Technology and Service Company. One step RT-PCR kit was bought from Qiagen (Germany). All other reagents were of the highest grade commercially available.
Vascular endothelial growth factor (vegf)
Manufactured by Sangon
Sourced in China
VEGF is a protein that promotes the growth of new blood vessels. It is a key regulator of angiogenesis, the process of forming new blood vessels from existing ones. VEGF is essential for the development and maintenance of the vascular system.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (1 mentions)
Hepg2 cell, by American Type Culture Collection (1 mentions)
Penicillin, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Onestep rt pcr kit, by Qiagen (1 mentions)
Gadph, by Sangon (1 mentions)
Mmp 9, by Sangon (1 mentions)
Bcl 2, by Sangon (1 mentions)
Lightcycler 96 system, by Roche (1 mentions)
Sybr green premix pro taq hs qpcr kit, by Accurate Biology (1 mentions)
Evo m mlv rt premix for qpcr, by Accurate Biology (1 mentions)
Mircute plus mirna first strand cdna synthesis kit, by Tiangen Biotech (1 mentions)
Reverse transcription reagent kit, by Takara Bio (1 mentions)
Mir 138 5p, by RiboBio (1 mentions)
Sybr green mix, by Takara Bio (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Gapdh, by Sangon (1 mentions)
Notch1, by Sangon (1 mentions)
Thrombin, by Merck Group (1 mentions)
Synergy h4, by Agilent Technologies (1 mentions)
4 protocols using vascular endothelial growth factor (vegf)
1
Quantitative Analysis of Sodium New Houttuyfonate in HepG2 Cells
2
Quantitative Analysis of M1/M2 Macrophage Markers
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Total RNA was extracted by Trizol following the manufacturer’s instructions, precipitated with isopropanol, and washed with ethanol. It was then converted to cDNA using Evo M-MLV RT Premix for qPCR (Accurate Biotechnology, Hunan, China) and analyzed by RT-PCR with a Roche LightCycler 96 system (Roche, Basel, Switzerland) using the SYBR Green Premix Pro Taq HS qPCR Kit (Accurate Biology). The following primers were used in this experiment: TNFA, CCL2 (for M1), CCL18, PPAR γ, CD206 (for M2), KLF4, VEGF, and human ACTB (Sangon Biotech, Shanghai, China, B661102-0001). All the primer sequences used in this investigation are listed in Table 1 . The expression of each gene was assessed by comparing it to the expression of β-actin using the 2−ΔΔCt method.
3
Profiling Gene and miRNA Expression in ADMSCs
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Total RNA was isolated from ADMSCs using the TRIzol (Invitrogen, USA) extraction method. cDNA libraries were synthesized from extracted mRNA using the Reverse Transcription Reagent kit (Takara, Japan) and random primers, while the cDNA library of miRNA was synthesized using the miRcute Plus miRNA First-Strand cDNA synthesis Kit (TIANGEN, China). Samples were submitted for Real-Time PCR using SYBR Green mix (Takara, Japan), and results were normalized to Gapdh (for mRNA) or U6 (for miRNA) expression levels. Gapdh, Cd31, Vegf, and Notch-1 specific primers were obtained from Sangon Biological Engineering (Shanghai, China), while miR-1956, miR-192-3p, miR-138-5p, and U6 specific primers were obtained from RiboBio. Primer sequences are presented in Table S2 . Data were analyzed through the comparative Ct (ΔΔCt) method to quantify relative gene expression.
4
Peptide-Based Fluorescent Biosensor
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The peptide sequence, RRHFFGCEKEKEKEPPPPC [12 (link),37 (link)], was obtained from Sangon (Shanghai, China) Co., Ltd. The chymotrypsin was obtained from Shanghai Linc-Bio Science (Shanghai, China) Co., Ltd. Cu(NO3)2, His, Ascorbic Acid, NaBH4, and other chemicals were obtained from the National Pharmaceutical Group Chemical Reagent (Beijing, China) Co., Ltd. PCN was obtained from Xi’an Qiyue Biotechnology(Xi’an, China) Co., Ltd. Thrombin, Lysozyme, N-Hydroxysuccinimide (NHS), N-(3-Dimetylaminopropyl)-N′-etylcarbodiimid (EDC), IgG, and BSA were obtained from Sigma-Aldrich (Shanghai, China). The serum was bought from Tianhang Biotechnology Co., Ltd. (Zhejiang, China). VEGF, a tubular dialyzer, and a 0.22 μm filter were obtained from Shanghai Sangon Co., Ltd. The intensity of the fluorescence was obtained using a Bio-Tek Synergy H4 multifunctional microplate reader.
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