C 22011

Manufactured by PromoCell

C-22011 is a laboratory centrifuge designed for general-purpose applications. It has a maximum speed of 6,000 rpm and a maximum relative centrifugal force (RCF) of 4,370 x g. The centrifuge can accommodate a variety of sample volumes and tube sizes.

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Lab products found in correlation

Pump system, by Ibidi (2 mentions) Type 1 collagen, by BD (2 mentions) C 12271, by PromoCell (2 mentions) Cc 3516, by PromoCell (2 mentions) Cc 4176, by PromoCell (2 mentions) Slide 1 0.4 luer, by Ibidi (1 mentions) μ slide i0.4 luer, by Ibidi (1 mentions) C 22062, by PromoCell (1 mentions) C 12500, by PromoCell (1 mentions) C 12200, by PromoCell (1 mentions) Hek293t, by Takara Bio (1 mentions) C 12203, by PromoCell (1 mentions) C 23020, by PromoCell (1 mentions) Tissuelyser 2, by Qiagen (1 mentions) Protease inhibitor cocktail, by Sangon (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) Tnf α, by Thermo Fisher Scientific (1 mentions) Hek293t, by American Type Culture Collection (1 mentions)

6 protocols using c 22011

Shear Stress Conditioning of Aortic Endothelial Cells

Cited 1 time

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Human aortic endothelial cells (HAECs) were obtained commercially (GIBCO, C0065C or PromoCell, C-12271) and cultured according to the manufacturer’s standard protocol. HAECs were seeded at various density to meet the purposes of different studies, and maintained in EC medium containing growth supplements (EBM-2, Lonza, CC-3516, CC-4176 or PromoCell, C-22011) with 2% FBS. HAECs between passage 6 and 7 were used for experiments. For shear experiments, confluent HAECs in 100 mm dishes were exposed to steady laminar shear (LS, 15 dyn/cm²), or oscillatory shear (OS, ±5 dyn/cm²) conditions for 2 days using a cone-and-plate shear device as previously described [20 (link), 21 (link)]. For shear experiments with gain of function or loss of function components, the commercially available Ibidi pump system (Ibidi, Germany) was used to reduce reagent use, and set up according to manufacturer instructions. LS (15 dyn/cm²) or OS (±5 dyn/cm²) was applied for 2 days to 80% confluent HAECs cultured overnight on microchannel slides (µ-Slide I^0.4 Luer, Ibidi, Germany) coated with 40 μg/ml collagen (Collagen Type I, BD Biosciences 35–4236).

Wang Y., Sun H.Y., Kumar S., Puerta M.D., Jo H, & Rezvan A. (2018). ZBTB46 is a shear-sensitive transcription factor inhibiting endothelial cell proliferation via gene expression regulation of cell cycle proteins. Laboratory Investigation; a Journal of Technical Methods and Pathology, 99(3), 305-318.

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Human Aortic Endothelial Cell Shear Experiments

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Human aortic endothelial cells (HAECs) were obtained commercially (GIBCO, C0065C or PromoCell, C-12271) and cultured according to the manufacturer’s standard protocol. HAECs were seeded at various density to meet the purposes of different studies, and maintained in endothelial cell medium containing growth supplements (EBM-2, Lonza, CC-3516, CC-4176 or PromoCell, C-22011) with 2% FBS. HAECs between passage 6 and 7 were used for experiments. For shear experiments, confluent HAECs in 100 mm dishes were exposed to steady laminar shear (LS, 15dyn/cm²), or oscillatory shear (OS, ±5dyn/cm²) conditions for 2 days using a cone-and-plate shear device as previously described.^{20 (link), 21 (link)} For shear experiments with gain of function or loss of function components, the commercially available Ibidi pump system (Ibidi, Germany) was used to reduce reagent use, and set up according to manufacturer instructions. Steady laminar shear (LS, 15dyn/cm²) or oscillatory shear (OS, ±5dyn/cm²) was applied to 80% confluent HAECs cultured overnight on microchannel slides (μ-Slide I^0.4 Luer, Ibidi, Germany) coated with 40μg/ml collagen (Collagen Type I, BD Biosciences 35-4236).

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Vascular cell adhesion on PCL films

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Primary human umbilical vein endothelial cells (HUVECs; C-12200 PromoCell GmbH, Heidelberg, DEU) and primary human umbilical artery smooth muscle cells (HUASMCs; C-12500 PromoCell GmbH, Heidelberg, DEU), were used to assess vascular cell adhesion onto the PCL film samples. HUVECs were cultured in endothelial cell growth medium (C-22011 PromoCell GmbH, Heidelberg, DEU ) supplemented with 1% (v/v) penicillin-streptomycin, 2% (v/v) fetal bovine serum, 0.4% (v/v) endothelial cell growth supplement, 0.1 ng/ml EGF, 1 ng/ml bFGF, 90 μg/ml heparin, and 1 μg/ml hydrocortisone. HUASMCs were cultured in smooth muscle cell growth medium (C-22062 PromoCell GmbH, Heidelberg, DEU) supplemented with 1% (v/v) antibiotic-antimycotic, 5% (v/v) fetal bovine serum, 0.5 ng/ml EGF, 2 ng/ml bFGF, and 5 μg/ml insulin. All cells were incubated at 37 °C, 5% CO₂ and grown to 80% confluency before use with experiments. Only passages between 3 and 6 were used for both cell lines.

Ammann K.R., Li M., Hossainy S, & Slepian M.J. (2019). The Influence of Polymer Processing Methods on Polymer Film Physical Properties and Vascular Cell Responsiveness. ACS applied bio materials, 2(8), 3234-3244.

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Cultivation of Primary Human Endothelial Cells

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Primary HUVECs (Promocell, C-12203) were cultured in endothelial cell growth medium 2 (EGM2) with a supplemental mix (Promocell, C-22011) and were used at passages 3–8. EA.hy926 (ATCC, CRL-2922), HEK-293T (Clontech, 632180), and NIH-3T3 (ATCC, CRL-1658) cells were cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (FBS). All cells were maintained in a humidified atmosphere at 37°C and 5% CO₂.

Zhang Y., Kong Y., Guo H., Liu Y., Zang Y, & Li J. (2022). Inner nuclear membrane protein TMEM201 maintains endothelial cell migration and angiogenesis by interacting with the LINC complex. Journal of Molecular Cell Biology, 14(3), mjac017.

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Quantifying Tumor Cytokines in Tissues

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To measure cytokines in tumor lysate, peritumor connective tissue and adipose tissue were carefully removed from tumors. Then tumors were quickly homogenized using the TissueLyser II (Qiagen) to ensure uniformity. Lysate was prepared using 1× PBS supplemented with a protease inhibitor cocktail (1:100, B14001; Sangon). following centrifugation, the supernatants were used for quantifying the concentration of MIF and active TGF-β1 with MIF ELISA kit (JL11079; Jianglaibio, China) and TGF-β1 ELISA kit (JL12223; Jianglaibio, China) according to the manufacturer’s instructions.
To measure cytokines in cell supernatants, isolated CAFs and CD146⁺ cells were cultured in fibroblast growth medium 2 (C-23020, PromoCell) supplemented with 1% penicillin/streptomycin (15070063, Gibco™) and in EC growth medium 2 (C-22011, PromoCell) supplemented with 1% penicillin/streptomycin (15070063, Gibco™), respectively. After 48 h, the supernatants of CAFs and CD146⁺ cells were collected and subjected for ELISA analysis using MIF ELISA kit (JL11079; Jianglaibio, China) and active TGF-β1 ELISA kit (JL12223; Jianglaibio, China) according to the manufacturer’s instructions.

Zhu J., Yang W., Ma J., He H., Liu Z., Zhu X., He X., He J., Chen Z., Jin X., Wang X., He K., Wei W, & Hu J. (2024). Pericyte signaling via soluble guanylate cyclase shapes the vascular niche and microenvironment of tumors. The EMBO Journal, 43(8), 1519-1544.

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Culturing and Activating Endothelial Cells

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HUVECs were cultured on fibronectin (FN)-coated dishes, using Endothelial Basal Medium, supplemented with singlequots (Promocell, C-22011) and 100 U/mL penicillin and streptomycin (P/S) at 37ᵒC in 5% CO₂ to create Endothelial Cell Growth Medium 2 (EGM-2). HUVECs were used between passage 4 and 7 and were never grown to grow above 70% confluency until a monolayer was required for an experiment. BOECs were cultured on 0.1% gelatin-coated dishes. For BOECs, 1:5 fetal calf serum (FCS) (Bodinco, Alkmaar, The Netherlands) was added to EGM-2, creating EGM-18 medium. To mimic inflammation in HUVECs or BOECs, TNFα (Peprotech, 300-01A) was added 10 ng/mL overnight to induce inflammatory protein expression in ECs. HEK-293T were purchased from ATCC (CRL-3216) and were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Gibco, 41965-039) containing 10% fetal calf serum, 100 U/mL P/S. Primary human neutrophils were collected from peripheral blood extracted from healthy voluntary donors, employed at Sanquin in The Netherlands, that signed informed consent according to the rules maintained by the Sanquin Medical Ethical Committee, which are based on rules and legislation in place within The Netherlands.

Grönloh M.L., Arts J.J., Mahlandt E.K., Nolte M.A., Goedhart J, & van Buul J.D. (2023). Primary adhered neutrophils increase JNK1-MARCKSL1-mediated filopodia to promote secondary neutrophil transmigration. iScience, 26(8), 107406.

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