Human Epidermal Keratinocytes from neonates (HEKn) (from Dr. Wendy Havran) or a mouse keratinocyte cell line, PAM212, were stimulated with 20 ng/ml of recombinant human LIGHT (R&D, 664-LI/CF) or mouse LIGHT for 72h in Epilife media (Life technologies). TSLP was measured by immunostaining, using anti-hTSLP mAb (clone AF1398, from R&D) or anti-mTSLP (clone 18, Santa Cruz Biotechnology), and qPCR analyses. For TGF-β neutralization, 30 μg/ml of anti-TGF-β mAb (1D11) or its Isotype control were added into culture. LIGHT receptors were visualized using anti-LTβR and anti-HVEM (Biolegend).
664 li cf
Manufactured by R&D Systems
The 664-LI/CF is a lab equipment product from R&D Systems. It is designed to perform a core function, but a detailed description while maintaining an unbiased and factual approach is not available.
Automatically generated - may contain errors
Lab products found in correlation
Epilife media, by Thermo Fisher Scientific (3 mentions)
Anti hvem, by BioLegend (3 mentions)
Anti htslp mab clone af1398, by R&D Systems (2 mentions)
Anti mtslp clone 18, by Santa Cruz Biotechnology (2 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
Opti mem medium, by Thermo Fisher Scientific (1 mentions)
Qiashredder, by Qiagen (1 mentions)
Hiperfect transfection reagent, by Qiagen (1 mentions)
On targetplus sirna, by Horizon Discovery (1 mentions)
Anti tgf β mab 1d11, by BioXCell (1 mentions)
4 protocols using 664 li cf
1
LIGHT-Induced TSLP Expression in Keratinocytes
2
RA-FLS Gene Expression Analysis
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Individual RA-FLS cell lines (2 × 106 cells/well of primary cultured RA-FLS) from patients 1–4 were incubated in either 1,000 ng/mL of recombinant human LIGHT protein (#664-LI/CF, R&D Systems, Minneapolis, MN) or OPTI-MEM medium (#31985-070, Thermo Fisher Scientific, Inc., Waltham, MA) for 12 h at 37 °C in 5% CO2. QIAshredder (#79,654, Qiagen, Hilden, Germany) and an RNeasy Mini kit (#74,104, Qiagen) were used to extract RNA from each cultured RA-FLS cell line, according to the manufacturer’s instructions.
Gene expression was detected using a microarray assay (Human Genome U133 Plus 2.0, GeneChip® 3’ Expression Array, Thermo Fisher Scientific), according to the manufacturer’s instructions.
Gene expression was detected using a microarray assay (Human Genome U133 Plus 2.0, GeneChip® 3’ Expression Array, Thermo Fisher Scientific), according to the manufacturer’s instructions.
3
LIGHT-Induced TSLP Expression in Keratinocytes
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Human Epidermal Keratinocytes from neonates (HEKn) (from Dr. Wendy Havran) or a mouse keratinocyte cell line, PAM212, were stimulated with 20 ng/ml of recombinant human LIGHT (R&D, 664-LI/CF) or mouse LIGHT for 72h in Epilife media (Life technologies). TSLP was measured by immunostaining, using anti-hTSLP mAb (clone AF1398, from R&D) or anti-mTSLP (clone 18, Santa Cruz Biotechnology), and qPCR analyses. For TGF-β neutralization, 30 μg/ml of anti-TGF-β mAb (1D11) or its Isotype control were added into culture. LIGHT receptors were visualized using anti-LTβR and anti-HVEM (Biolegend).
4
LIGHT and IL-13 Stimulate Periostin Expression
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Human epidermal keratinocytes from neonates (HEK) or mouse epidermal keratinocytes (PAM212 cell line) were stimulated with 20 ng/ml recombinant human LIGHT (664-LI/CF; R&D), mouse LIGHT (1794-LT-025/CF; R&D), or mouse IL-13 (413-ML-005; R&D) for 72 h in Epilife media (Life Technologies). In some cases, 30 µg/ml anti–TGF-β mAb (1D11; BioXCell) or anti–IL-13 (MAB413-SP; R&D) or isotype control were added into culture. Periostin was measured by immunostaining using anti–hPeriostin mAb (clone ABT292; Millipore) and quantitative PCR analyses. ON-TARGETplus siRNAs were purchased from Dharmacon. A 5–25 nM concentration range of control scrambled or HVEM siRNA was transfected into HEK cells using HiPerFect transfection reagent (Qiagen). Efficiency of HVEM knockdown was monitored over time by both mRNA and surface protein levels and was stable during the 24- to 72-h culture period. HVEM was visualized using anti-HVEM (BioLegend).
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