All human primary vascular ECs (HAEC, HCAEC, HDMEC, HLMEC, and HUVEC) were purchased from Lonza Walkersville Inc., cultured in EGM or EGM2 medium according to the manufacturer instruction, and used for experiment in less than five passages. The human acute monocytic leukemia cell line THP-1 was obtained from American Type Culture Collection (ATCC) and maintained in RPMI 1640 medium (Corning) containing 10% FBS (Sigma-Aldrich). HeLa, A549, RAW264.7, and U937 were purchased from ATCC and cultured in Dulbecco's modified Eagle's medium supplemented with 10% FBS. Transient transfection of TRIM14 vector and siRNA into HUVECs was performed by electroporation using Nucleofector device (Lonza) and Nucleofector kits for HUVEC (Lonza) following the manufacturer’s instruction. After electroporation, cells were plated into 35 mm dishes and incubated for 24 h at 37°C, 5% CO2. Then, cells were treated with 10 ng/ml TNF-α or IL-1β for indicated times, and proteins from those cells were extracted and detected by western blot.
Nucleofector kits for huvec
Manufactured by Lonza
The Nucleofector kits for HUVEC are a set of specialized transfection reagents designed for efficient delivery of plasmid DNA, siRNA, or other macromolecules into human umbilical vein endothelial cells (HUVEC). The kits provide optimized protocols and solutions to facilitate the transfection process.
Automatically generated - may contain errors
Lab products found in correlation
Hdmec, by Lonza (3 mentions)
Rpmi 1640 medium, by Corning (3 mentions)
Nucleofector device, by Lonza (2 mentions)
Human umbilical vein endothelial cells (huvec), by Lonza (2 mentions)
Raw264, by American Type Culture Collection (2 mentions)
Thp 1, by American Type Culture Collection (2 mentions)
Hcaec, by Lonza (2 mentions)
Hlmec, by Lonza (2 mentions)
Huvecs, by Lonza (1 mentions)
Egm endothelial cell growth medium bulletkit, by Lonza (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Hek293t cell, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
2 mercaptoethanol, by Merck Group (1 mentions)
Egm 2 endothelial cell growth medium 2 bulletkit, by Lonza (1 mentions)
Nucleofactor device, by Lonza (1 mentions)
3 protocols using nucleofector kits for huvec
1
Endothelial Cell Stimulation Assay
2
Endothelial Cell Culture and Transfection
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All human primary vascular ECs including human aortic ECs (HAECs), human coronary artery ECs (HCAECs), human dermal microvascular ECs (HDMECs), human lung microvascular ECs (HLMECs), and human umbilical vein ECs (HUVECs) were purchased from Lonza Walkersville Inc. maintained in EGM™ Endothelial Cell Growth Medium BulletKit™ or EGM™-2 Endothelial Cell Growth Medium-2 BulletKit™ according to the manufacturer's instruction. For the present study, cells less than passage five were used for the experiment. THP-1 cells (human acute monocytic leukemia cell) were obtained from ATCC and cultured in 10% FBS (Sigma-Aldrich) RPMI 1640 medium (Corning), 1% penicillin/streptomycin (Invitrogen), and 0.05 mmol/L 2-mercaptoethanol (Sigma Aldrich). HEK293T cells were bought from ATCC and seeded in Dulbecco's modified Eagle's medium supplemented with 10% FBS and 1% penicillin/streptomycin (Invitrogen). Transfection of Ataxin-10 vector and siRNA into HUVECs transiently was performed by electroporation using Nucleofector device and Nucleofector kits for HUVEC (Lonza) as described [22 (link), 23 (link)]. After 24 h, 10 ng/mL TNF-α was added to the cells for indicated times, and cell lysates were collected and detected by Western blotting.
3
Endothelial Cell Stimulation by TNFα
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All human primary vascular endothelial cells (HAEC, HCAEC, HDMEC, HLMEC, and HUVEC) were purchased from Lonza Walkersville Inc., cultured in EGM or EGM2 medium according to the manufacturer instruction, and used for experiment in less than five passages. The human acute monocytic leukemia cell line THP-1 was obtained from American Type Culture Collection (ATCC) and maintained in RPMI 1640 medium (Corning) containing 10% FBS (Sigma-Aldrich). HeLa, A549, RAW264.7, and U937 were purchased from ATCC and cultured in DMEM supplemented with 10% FBS. Transient transfection of TRIM65 vector and siRNA into HUVECs was performed by electroporation using Nucleofactor device (Lonza) and Nucleofector kits for HUVEC (Lonza) following the manufacturer’s instruction. After electroporation, cells were plated into 35 mm dishes and incubated for 24 h at 37°C, 5% CO2. Then, cells were treated with 10 ng/ml TNFα for indicated times and proteins from those cells were extracted and detected by western blotting.
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