Topcount luminescence counter

Manufactured by PerkinElmer

Sourced in United States

The TopCount Luminescence Counter is a high-performance microplate reader designed for luminescence-based assays. It provides accurate and reliable detection of light signals generated in 96- or 384-well microplates. The instrument's core function is to quantify light emission, enabling researchers to analyze a wide range of luminescent samples, including cell-based assays, reporter gene assays, and biochemical reactions.

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Lab products found in correlation

Pgn sa, by InvivoGen (2 mentions) Beta glo reagent, by Promega (2 mentions) Pgn bs, by InvivoGen (2 mentions) Pgn ec, by InvivoGen (2 mentions) Celltiter glo luminescent cell viability assay, by Promega (1 mentions) Enduren live cell luciferase substrate, by Promega (1 mentions)

Close spelling variants of this product

TopCount (96 citations) Luminescence Counter (26 citations) TopCount NXT Microplate Scintillation and Luminescence Counter (18 citations) TopCount NXT Scintillation and Luminescence Counter (6 citations) TopCount microplate scintillation and luminescence counter (6 citations) TopCount NXT microplate luminescence counter (5 citations) TopCount NXT Microplate Scintillation & Luminescence Counter (5 citations) Packard TopCount NXT Microplate Scintillation and Luminescence Counter technology (2 citations)

5 protocols using topcount luminescence counter

PGLYRP1 Activation of TREM-1 Signaling

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Example 9

To test if recombinant PGLYRP1 can activate a TREM-1 response, the BWZ/hTREM-1 reporter cell line was seeded into black 96 well plates and stimulated with recombinant human PGLYRP1 (Cat, no. 2590-PG-050, R&D Systems: Minneapolis, Minn.) in the presence or absence of 10 μg/ml PGN. TREM-1 activation was read out after 24 hours of culture using the BetaGlo reagent (Cat. no. E4720, Promega Madison, Wis., USA) and luminescence measured using a TopCount Luminescence counter from Perkin Elmer. As shown in FIG. 5A, stimulation of the TREM-1 reporter cell line with PGLYRP1 induced a dose-dependent activation of TREM-1 in the presence of PGN. Several different PGN were tested, including PGN-EC (from E. coli), PGN-SA (from S. aureus) and PGN-BS (from B. subtilis) (Invivogen, San Diego, Calif., USA), and were all able to fascilitate the PGLYRP1-induced TREM-1 response.

The response induced by recombinant PGLYRP1 could be inhibited by addition of a recombinant TREM-1-Fc-fusion protein (SEQ ID NO: 2) (FIG. 5B) or by addition of polyclonal anti-PGLYRP1 antibody (Cat. no. AF-2590, R&D Systems: Minneapolis, Minn., USA) confirming that PGLYRP1 is the TREM-1 ligand and that soluble TREM-1 and anti-PGLYRP1 are potentially useful as TREM-1 antagonists.

US10906965B2. Methods of treating autoimmune disease or chronic inflammation wtih antibodies that bind peptidoglycan recognition protein 1 (2021-02-02). Novo Nordisk A/S [DK]. Inventors: Vibeke Westphal Stennicke [DK], Christine Brender Read [DK], Joseph Leon Kuijper [US], Xiaoting Tang [US], Mark Heipel [US], Siv Annegrethe Hjorth [DK].

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Cell Proliferation Assay with CellTiter-Glo

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Cell proliferation was measured using CellTiter-Glo® Luminescent Cell Viability Assay (Promega, Madison, WI), following manufacturer’s instructions. Briefly, 1000 EG7 cells were seeded in a total volume of 200 μl of RPMI + 10% FBS per well in a 96-well plate. The next day, compound plates were reconstituted in complete media with a 10 point dose curve. Cell media of the plate was aspirated and compound with a total volume of 100 μl was incubated with the cells for 72 hours. 100 μl of CellTiter-Glow reagent was added to each well and protected from the light with a plate seal. Plates were shaken at room temperature for 2 minutes to induce cell lysis and then allowed to sit for approximately 10 minutes. The raw data was measured on a TopCount Luminescence Counter (Perkin Elmer, Waltham, MA) and analyzed in Prism using an [inhibitor] vs. response – variable slope (four parameters) equation.

Huarte E., O’Connor R.S., Peel M.T., Nunez-Cruz S., Leferovich J., Juvekar A., Yang Y.O., Truong L., Huang T., Naim A., Milone M.C, & Smith P.A. (2020). Itacitinib (INCB039110), a JAK1 inhibitor, Reduces Cytokines Associated with Cytokine Release Syndrome Induced by CAR T-Cell Therapy. Clinical cancer research : an official journal of the American Association for Cancer Research, 26(23), 6299-6309.

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PGLYRP1 Activates TREM-1 in the Presence of PGN

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Example 9

To test if recombinant PGLYRP1 can activate a TREM-1 response, the BWZ/hTREM-1 reporter cell line was seeded into black 96 well plates and stimulated with recombinant human PGLYRP1 (Cat. no. 2590-PG-050, R&D Systems: Minneapolis, Minn.) in the presence or absence of 10 μg/ml PGN. TREM-1 activation was read out after 24 hours of culture using the BetaGlo reagent (Cat. no. E4720, Promega Madison, Wis., USA) and luminescence measured using a TopCount Luminescence counter from Perkin Elmer. As shown in FIG. 5A, stimulation of the TREM-1 reporter cell line with PGLYRP1 induced a dose-dependent activation of TREM-1 in the presence of PGN. Several different PGN were tested, including PGN-EC (from E. coli), PGN-SA (from S. aureus) and PGN-BS (from B. subtilis) (Invivogen, San Diego, Calif., USA), and were all able to fascilitate the PGLYRP1-induced TREM-1 response.

US10150809B2. Antibodies that bind peptidoglycan recognition protein 1 (2018-12-11). Novo Nordisk A/S [DK]. Inventors: Vibeke Westphal Stennicke [DK], Christine Brender Read [DK], Joseph Leon Kuijper [DK], Xiaoting Tang [DK], Mark Heipel [DK], Siv Annegrethe Hjorth [DK].

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Dual-Split Protein Cell-Cell Fusion Assay for RSV F Protein

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The dual-split protein (DSP) reporter cell-cell fusion assay was previously adapted to measuring RSV F protein activity [28 (link), 33 (link), 34 (link)]. 293T “effector” (cis) cells were transfected with RSV A2 F or 2–20 F and DSP_1-7 in the presence of fusion inhibitor BMS-433771 (a gift from Jin Hong, Alios Biopharma, San Francisco, CA). 293T “target cells” (trans) were transfected with DSP_8-11 and pCG-SLAM or pcDNA3.1 empty vector. Effector cis or target trans cells were transfected with pTriex3-EGFR. Effector and target cells were washed with PBS 24 h post-transfection and harvested by pipetting in media containing EnduRen live cell luciferase substrate (Promega). Equal volumes of effector and target cells were mixed and placed into an opaque 96-well plate in quadruplicate. Plates were incubated at 37°C and luciferase activity as a measure of cell-cell fusion was assayed on a TopCount Luminescence counter (Perkin Elmer) 4, 6, and 8 h after cell mixing. A positive control of DSP_1-7 and DSP_8–11 transfected into the same cell population was used to validate replicates.

Currier M.G., Lee S., Stobart C.C., Hotard A.L., Villenave R., Meng J., Pretto C.D., Shields M.D., Nguyen M.T., Todd S.O., Chi M.H., Hammonds J., Krumm S.A., Spearman P., Plemper R.K., Sakamoto K., Peebles RS J.r., Power U.F, & Moore M.L. (2016). EGFR Interacts with the Fusion Protein of Respiratory Syncytial Virus Strain 2-20 and Mediates Infection and Mucin Expression. PLoS Pathogens, 12(5), e1005622.

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Luminescent E. coli Quantification

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E. coli (50 μl, OD₆₀₀ = 0.5) suspended in PBS were added into 96-well white plates and luminescence was detected on a Top Count Luminescence Counter (Perkin-Elmer Life Sciences, Waltham, MA).

Hsieh Y.T., Chen K.C., Cheng C.M., Cheng T.L., Tao M.H, & Roffler S.R. (2015). Impediments to Enhancement of CPT-11 Anticancer Activity by E. coli Directed Beta-Glucuronidase Therapy. PLoS ONE, 10(2), e0118028.

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