Hek blue reporter cells

Manufactured by InvivoGen

HEK-Blue™ reporter cells are a cell line designed to monitor the activation of specific signaling pathways. These cells stably express a secreted embryonic alkaline phosphatase (SEAP) reporter gene under the control of an inducible promoter. The reporter gene activation can be easily measured in the cell culture supernatant, providing a quantitative readout of the signaling pathway of interest.

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Lab products found in correlation

Polymixin b, by InvivoGen (1 mentions) Neutralizing antibodies, by InvivoGen (1 mentions)

3 protocols using hek blue reporter cells

Evaluating Innate Immune Receptor Activation

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Human (h)TLR-2/−3/−4/−5/−7/−8/−9/Null, NOD-1/−2, and Dectin1b-specific HEK-Blue™ reporter cells (InvivoGen, San Diego, CA), were grown, maintained and utilized as specified by the manufacturer (InvivoGen, San Diego, CA). To establish response ratio curves of natural ligands (PAMPs) of each PRR we used positive controls (Table 2) according to the manufacturer’s instructions (InvivoGen, San Diego, CA). Polymixin B (InvivoGen, San Diego, CA) was used in all experiments to eliminate interference from endogenous LPS. (See Supplemental Methods for further description).

Sosa R.A., Rossetti M., Naini B.V., Groysberg V.M., Kaldas F.M., Busuttil R.W., Chang Y.L., Gjertson D.W., Kupiec-Weglinski J.W, & Reed E.F. (2018). Pattern Recognition Receptor-Reactivity Screening of Liver Transplant Patients: Potential for Personalized and Precise Organ Matching to Reduce Risks of Ischemia-Reperfusion Injury. Annals of surgery.

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Blocking TLR4 Signaling in HEK-Blue Cells

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Human (h)TLR4-specific HEK-Blue™ reporter cells (InvivoGen, San Diego, CA), were grown, maintained and utilized as previously described (3 ). Neutralizing antibodies (InvivoGen) were added for one hour prior to cell stimulation with patient LF to block the function of TLR4, CD14, and/or MD2.

Sosa R.A., Terry A.Q., Kaldas F.M., Jin Y.P., Rossetti M., Ito T., Li F., Ahn R.S., Naini B.V., Groysberg V.M., Zheng Y., Aziz A., Nevarez-Mejia J., Zarrinpar A., Busuttil R.W., Gjertson D.W., Kupiec-Weglinski J.W, & Reed E.F. (2020). Disulfide-HMGB1 Drives Ischemia-Reperfusion Injury in Human Liver Transplantation. Hepatology (Baltimore, Md.), 73(3), 1158-1175.

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Serum-Based Immune Response Assay

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Blood was collected post-mortem via cardiac puncture and incubated at room temperature for 30 minutes in 1.5ml tubes to allow clotting. Samples were then centrifuged at 1,500g for 20 minutes at 4°C after which serum was collected to new tubes. 20μl of serum was added to wells containing InvivoGen HEK-Blue™ reporter cells, and luminal antigens were detected following manufacturer’s instructions.

Naama M., Telpaz S., Awad A., Ben-Simon S., Harshuk-Shabso S., Modilevsky S., Rubin E., Sawaed J., Zelik L., Zigdon M., Asulin N., Turjeman S., Werbner M., Wongkuna S., Feeney R., Schroeder B.O., Nyska A., Nuriel-Ohayon M, & Bel S. (2023). Autophagy controls mucus secretion from intestinal goblet cells by alleviating ER stress. Cell Host & Microbe, 31(3), 433-446.e4.

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