To evaluate the effect of UV irradiation (290–320 nm), a type B ultraviolet lamp was used. Phage samples (107 PFU/mL) were placed directly on the TL 20 W/12 RS lamp (Philips, Holland). The experiments were carried out in phage buffer at pH 7.0 and at room temperature. Samples were exposed for 10, 20, 30, 40, 60 or120 min and serial dilutions were prepared. They were plated in triplicate (technical repeats) for incubation at 25 °C. The average number of lysis ranges and the standard deviation were calculated.
Tl20w 12rs lamp
Manufactured by Philips
Sourced in Germany, Netherlands
The TL20W/12RS lamp is a fluorescent lamp designed for laboratory and industrial applications. It provides general illumination and is suitable for use in various lighting fixtures. The lamp has a power rating of 20 watts and a color temperature of 12,000 Kelvin, which produces a cool, bright light. The TL20W/12RS lamp is a reliable and energy-efficient lighting solution for professional environments.
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Lab products found in correlation
Uvx 31 sensor, by Analytik Jena (1 mentions)
Roscovitine, by Enzo Life Sciences (1 mentions)
Wortmannin, by Merck Group (1 mentions)
Epidermal growth factor (egf), by Merck Group (1 mentions)
Pd153035, by Merck Group (1 mentions)
Su9516, by Bio-Techne (1 mentions)
Bs 181, by Selleck Chemicals (1 mentions)
Ac devd cho, by Enzo Life Sciences (1 mentions)
Pd98059, by Merck Group (1 mentions)
Mg132, by Merck Group (1 mentions)
Humedia kg2, by Kurabo (1 mentions)
Skh 1, by Orient Bio (1 mentions)
Epcam g8.8 pecy7, by BioLegend (1 mentions)
Tnf α 315 01a, by Thermo Fisher Scientific (1 mentions)
Il 1β 211 11b, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 647, by Thermo Fisher Scientific (1 mentions)
7 protocols using tl20w 12rs lamp
1
UV Inactivation of Phage Samples
2
UVB Lamp Protocol for Research
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The UVB source employed in this study was a Phillips TL20W/12RS lamp (Phillips, Eindhoven, Holland). The energy exposed was measured using a UVX radiometer with a UVX-31 sensor (UVP Inc., San Gabriel, CA).
3
UVB-Induced Stress Response in HaCaT Cells
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HaCaT KC were provided by P. Boukamp (DKFZ/IUF) and authenticated by the German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany). The cultivation of HaCaT KC and the generation and cultivation of HaCaT-EV and HaCaT-shAHR KC has been previously described [10 (link)]. The source for UVB irradiation was a TL20W/12RS lamp (Philips, Eindhoven, The Netherlands), which emits most of its energy in the UVB range (290–320 nm) with an emission peak at 310 nm. For both UVB and sham exposure, culture medium was replaced by PBS. For cell treatment, Wortmannin, PD153035, PD98059, MG-132, BaP (all from Sigma-Aldrich, Munich, Germany), roscovitine (Enzo Life Sciences, Loerrach, Germany), BS-181 (Selleckchem, Houston, TX, USA), SU9516 (Tocris Bioscience, Bristol, UK) and MNF (provided by I. Meyer, Symrise AG, Holzminden, Germany) were dissolved in DMSO. EGF (Sigma-Aldrich) and Ac-DEVD-CHO (Enzo Life Sciences) were dissolved in water.
4
UVB Exposure of Normal Human Keratinocytes
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Normal human epidermal keratinocytes (NHEKs) were obtained from Kurabo (Osaka, Japan) and were cultured in HuMedia-KG2 (Kurabo) at 37℃ in a 5％ CO 2 atmosphere. NHEKs were exposed to 20 mJ/cm 2 UVB (TL20W/12RS lamp, Phillips, Hamburg, Germany) in Hank s buffer without Ca 2＋ and Mg 2＋ .
5
Hairless Mouse UV Irradiation Protocol
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All animal procedures (YWC-131205-1) were approved on 12 December 2013 by the Yonsei University Wonju Campus Institutional Animal Care and Use Committee (IACUC). Twenty-four female hairless mice (SKH-1) were purchased from Orient Bio (Seongnam, Korea) and were reared up to 18 weeks old in a standard environment with the temperature maintained at 22 ± 2 °C, relative humidity at 40% ± 5%, and a 12 h/12 h light and dark cycle. The mice were irradiated with a 1/2 minimal erythema dose (MED) (UVA: 14 J/cm2, UVB: 40 mJ/cm2) three times a week for 8 weeks. UVB and UVA radiation were delivered as previous reported [20 (link)]. Philips TL20W/12RS lamps (Philips, Utrecht, The Netherlands) and Philips CLEO performance 40 W lamps (Philips) have the wavelength of between 290 to 315 nm (UVB) and 315 to 380 nm (UVA), respectively.
6
Cytokine and Ultraviolet B Exposure Protocol
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TNF-α (315-01A) or IL-1β (211-11B) recombinant mouse peptides were purchased from PeproTech (Rocky Hills, NJ) and were resuspended in PBS. For UVB experiments, we used two TL 20W/12RS lamps (Philips, Amsterdam, The Netherlands). We used UVB doses previously described as 20 mJ/cm2 for pKCs or experiments, (Mohammed et al., 2016 (link)).
DMBA (57-97-6; Sigma-Aldrich, St. Louis, MI) was applied at 0.1mM in DMSO for pKC experiments. For in vivo mouse experiments, 10 mM of DMBA was resuspended in DMSO-to-ethanol-to-glycerol solution (1:1:3) and was applied to the flank skin of shaven mice. Antibodies directly conjugated to different fluorophores were used for flow cytometry and immunofluorescence. Anti-CD11c(N418)-PerCp5.5, CD11b(M1/70)-PeCy7, I-A/I-E/MHCII (M5/114.15.2)-AF700, Langerin(4c7)-phycoerythrin, CD103(2E7)-AF647, CD45.2(104)-BV605, Sca-1(E13-161.7)-PerCp5.5, EpCAM(G8.8)-PeCy7, and CD34(HM34)-PE/Dazzle594 were purchased from BioLegend (San Diego, CA). Viability Dye eFluor 780 (eBioscience; Invitrogen, Carlsbad, CA) was used for live‒dead discrimination. Anti-ανβ6 (6.3g9) and anti-ανβ8 (C6D4)-phycoerythrin were kindly provided by Dean Sheppard and Stephen Nishimura, respectively. Anti-ανβ6 (6.3g9) was directly conjugated to Alexa Fluor 647 (A20186; Thermo Fischer Scientific, Waltham, MA).
DMBA (57-97-6; Sigma-Aldrich, St. Louis, MI) was applied at 0.1mM in DMSO for pKC experiments. For in vivo mouse experiments, 10 mM of DMBA was resuspended in DMSO-to-ethanol-to-glycerol solution (1:1:3) and was applied to the flank skin of shaven mice. Antibodies directly conjugated to different fluorophores were used for flow cytometry and immunofluorescence. Anti-CD11c(N418)-PerCp5.5, CD11b(M1/70)-PeCy7, I-A/I-E/MHCII (M5/114.15.2)-AF700, Langerin(4c7)-phycoerythrin, CD103(2E7)-AF647, CD45.2(104)-BV605, Sca-1(E13-161.7)-PerCp5.5, EpCAM(G8.8)-PeCy7, and CD34(HM34)-PE/Dazzle594 were purchased from BioLegend (San Diego, CA). Viability Dye eFluor 780 (eBioscience; Invitrogen, Carlsbad, CA) was used for live‒dead discrimination. Anti-ανβ6 (6.3g9) and anti-ανβ8 (C6D4)-phycoerythrin were kindly provided by Dean Sheppard and Stephen Nishimura, respectively. Anti-ανβ6 (6.3g9) was directly conjugated to Alexa Fluor 647 (A20186; Thermo Fischer Scientific, Waltham, MA).
7
Repeated UVB Exposure in Keratinocytes
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For UVB exposure studies, we use two TL 20 W/12 RS lamps (Philips, Eindhoven, Netherlands), emitting UVB peaking at 313 nm that were placed 15 cm above the cell culture flasks. The emitted radiation was checked at the bottom of each flask using a UV-meter (Waldmann, Villingen-Schwenningen, Germany). For repeated exposures to UVB, keratinocytes were seeded in flasks 2 days before and the cells were exposed to UVB 5 mJ/cm2 in a serum free medium without flask cover. This exposure was repeated 6 times and the time interval between exposures was 30 min. Normal control (NC) cells were submitted to the same conditions but with no exposure to UVB.
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