Lyec 1

Manufactured by InvivoGen

Lyec-1 is a laboratory equipment product designed for cell lysis and protein extraction. It utilizes a lysosome-based extraction method to effectively disrupt cell membranes and release cellular contents, including proteins, for subsequent analysis or purification.

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Lab products found in correlation

Ifn α2, by BioLegend (1 mentions) 3p hprna, by InvivoGen (1 mentions) Tlrl hprna, by InvivoGen (1 mentions) Plad b, by Bio-Techne (1 mentions) Tnf α, by BioLegend (1 mentions) Non targeting sirna, by Horizon Discovery (1 mentions) G3 ysd, by InvivoGen (1 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)

2 protocols using lyec 1

Cell-Based Cytokine Induction Assay

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Plad B was purchased from Tocris (6070), and spliceostatin analog PF-06437837 (thailanstatin A methyl ester) was synthesized by Pfizer as previously reported (22 (link)). Cells were seeded in 96-well plates at 5000 cells/well (4T1), 10,000 cells/well (A549, CT26, and MC38 HEK), and 20,000 cells/well (B16F10) overnight in 75 μl media. The next day, 75 μl of a 2× solution of the indicated compound was added to wells, and cells were incubated at 37 °C until the indicated time points. Assays with THP1 cells were performed similarly except plating cells (100,000 cells) immediately before the addition of indicated compounds. Supernatant was removed and used in subsequent assays. 3p-hpRNA (Invivogen; tlrl-hprna) was prepared as a 2× solution in complex with the transfection reagent LyoVec (Invivogen; lyec-1) according to the manufacturer's instructions. After complexing, 75 μl of the 2× concentrated 3p-hpRNA/LyoVec was added to cells until the indicated time points. Positive control cytokines used were IFN-α2 (Biolegend; 592702) and TNFα (Biolegend; 570102).

Chang A.Y., Zhou Y.J., Iyengar S., Pobiarzyn P.W., Tishchenko P., Shah K.M., Wheeler H., Wang Y.M., Loria P.M., Loganzo F, & Woo S.R. (2021). Modulation of SF3B1 in the pre-mRNA spliceosome induces a RIG-I-dependent type I IFN response. The Journal of Biological Chemistry, 297(5), 101277.

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Investigating STING and cGAS in Cancer Cells

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Pan08.13, HupT4, Capan-2, A549, HCC827, and A549 Dual cells were seeded in 6-well plates at 1×10⁵, 1×10⁵, 1.5×10⁵, 6×10⁴, 1×10⁵, and 6×10⁴ cells/well, respectively. 24 hours after seeding, cells were transfected with either 25 pmol non-targeting siRNA (Dharmacon, Lafayette, CO; D-001810–10-20), STING-specific siRNA (Dharmacon, L-024333–02-0005), or cGAS-specific siRNA (Dharmacon, L-015607–02-0005) using Lipofectamine RNAi Max (Invitrogen, 13778–075), according to manufacturer’s protocols. 24 hours after transfection, cells were treated with either 0.1% DMSO or 2 μM GSK3368712 for 4 days. DMSO-treated cells were then treated with 500 ng/mL of the cGAS agonist, G3-YSD (Invivogen, tlrl-ydna), in LyoVec (Invivogen, lyec-1) for 24 hours.

Fedoriw A., Shi L., O’Brien S., Smitheman K.N., Wang Y., Hou J., Sherk C., Rajapurkar S., Laraio J., Williams L.J., Xu C., Han G., Feng Q., Bedford M.T., Wang L., Barbash O., Kruger R.G., Hwu P., Mohammad H.P, & Peng W. (2022). Inhibiting Type I arginine methyltransferase activity promotes T cell-mediated antitumor immune responses. Cancer immunology research, 10(4), 420-436.

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