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Quanti blue

Manufactured by Thermo Fisher Scientific
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Quanti-Blue is a colorimetric detection assay that quantifies the levels of alkaline phosphatase (AP) activity in solution. It is a simple, rapid, and sensitive method for the detection and quantification of AP levels.

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Lab products found in correlation

Phorbol myristate acetate (pma), by Merck Group (2 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Corning (1 mentions) Dulbecco modified eagle medium (dmem), by Welgene (1 mentions) Normocin, by InvivoGen (1 mentions) Zeocin, by InvivoGen (1 mentions) Hek blue il 1β cells, by InvivoGen (1 mentions) Fluoxetine, by Merck Group (1 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions) Elisa microplate reader, by Agilent Technologies (1 mentions) Poly d lysine hydrobromide, by Merck Group (1 mentions) Pnf κb seap, by Takara Bio (1 mentions) Prism 7, by GraphPad (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Hek blue htlr4 cells, by Thermo Fisher Scientific (1 mentions) Quanti blue, by InvivoGen (1 mentions) Quanti luc, by InvivoGen (1 mentions)

Close spelling variants of this product

QUANTI-Blue™ medium (3 citations) Quanti-Blue reagent (3 citations) QUANTI-Blue™ assay (2 citations)

6 protocols using quanti blue

1

Analyzing IL-1β Neutralizing Activity

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To examine IL-1β neutralizing activity of TA, HEK-Blue IL-1β cells (InvivoGen, San Diego, CA, USA) were used. Cells were cultured in DMEM (WELGENE, Gyeongsangbuk-do, Korea) containing 10% fetal bovine serum (FBS, Corning, Corning, NY, USA), 100 units/ml penicillin, 100 mg/ml streptomycin (Invitrogen), 100 μg/ml Normocin (InvivoGen), and 100 μg/ml Zeocin (InvivoGen), and cultured at 37°C in 5% CO2 humidified incubator. Cells (5×104 cells/well) were seeded in a 96-well plate and stimulated with human recombinant IL-1β (10 ng/ml) with or without various concentrations of TA (0.78, 1.56, 3.125, 6.25, 12.5, 25, 50, or 100 μM). After a 24 h incubation, SEAP activity was detected using QUANTI-Blue™ (Invitrogen) and the OD at 620 nm. Cell viability was measured by analyzing the OD at 450 nm using D-Plus CCK (DONGIN LS, Gyeonggi-do, Korea).
Lee H.R., Jeong Y.J., Lee J.W., Jhun J., Na H.S., Cho K.H., Kim S.J., Cho M.L, & Heo T.H. (2023). Tannic acid, an IL-1β-direct binding compound, ameliorates IL-1β-induced inflammation and cartilage degradation by hindering IL-1β-IL-1R1 interaction. PLOS ONE, 18(4), e0281834.
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2

Extraction and Storage of FAEW

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The FAEW extract was prepared from commercial pills (Xiaoyao wan) purchased from Wanxi Pharmaceutical Company (Henan Province, China). They were dissolved in H2O: MeOH: DCM in a ratio of 1: 4: 5 for 3 days. A rotary evaporator was used to remove the solvents and the final extracts were stored at −20°C. Fluoxetine was purchased from Sigma-Aldrich (Steinheim, Germany). Tumor necrosis factor-α (TNF-α) was purchased from Sino Biological Inc. (Peking, China). Quanti-Blue was purchased from Invitrogen and prepared according to the instructions of the manufacture.
Hong C., Schüffler A., Kauhl U., Cao J., Wu C.F., Opatz T., Thines E, & Efferth T. (2017). Identification of NF-κB as Determinant of Posttraumatic Stress Disorder and Its Inhibition by the Chinese Herbal Remedy Free and Easy Wanderer. Frontiers in Pharmacology, 8, 181.
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3

NF-κB Transactivation Assay

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Transient transfection of a reporter plasmid, pNF-kB-SEAP (Clontech Laboratories, Inc., Palo Alto, CA, USA) was performed for 293T cells seeded at 1 × 104/well in a 96-well plate using Lipofectamine 3000 (Invitrogen, Carlsbad, MA, USA). One day after transfection, the cell medium was replaced with fresh DMEM and treated with ChondroT or GHJTY (0.3 and 0.1 mg/mL) for 4 h. The cells were treated overnight with PMA (Sigma Co., St. Louis, MO, USA) at a concentration of 1 ng/mL. The supernatants were incubated with QUANTI-Blue (Invitrogen, Carlsbad, MA, USA) for 2–4 h, and the absorbance was read at 630 nm with an ELISA microplate reader (ELx808).
Park J.U., Kim S.J., Na C.S., Choi C.H., Seo C.S., Son J.K., Kang B.Y, & Kim Y.R. (2016). Chondroprotective and anti-inflammatory effects of ChondroT, a new complex herbal medication. BMC Complementary and Alternative Medicine, 16, 213.
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4

NF-κB Transcriptional Activity Assay in 293T Cells

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293T cells were seeded into poly-D-lysine hydrobromide (Sigma, MO, United States) coated 96 well plate at 1.5 × 105 cells/mL, and then transient transfection of a reporter plasmid pNF-κB-SEAP (Clontech Laboratories, Palo Alto, CA, United States) was performed to cells with HilyMax for 4 h. After transfection, the cell medium was replaced with fresh DMEM overnight. Then cells were treated with ZCO at the concentration of 0.0025, 0.005, and 0.01% or the positive control Bay11-7082 (20 μM) for 2 h prior to stimulation with 3 ng/mL of PMA (Sigma, St. Louis, MO, United States) for 24 h. The supernatants (10 μL) were incubated with Quanti-Blue (100 μL) (Invitrogen, Carlsbad, MA, United States) for 1 h, and the absorbance was read at 630 nm with an ELISA microplate reader (BioTek, Winooski, VT, United States).
Guo R.H., Park J.U., Jo S.J., Ahn J.H., Park J.H., Yang J.Y., Lee S.S., Park M.J, & Kim Y.R. (2018). Anti-allergic Inflammatory Effects of the Essential Oil From Fruits of Zanthoxylum coreanum Nakai. Frontiers in Pharmacology, 9, 1441.
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5

NF-κB Activation in HEK-Blue Cells

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HEK-Blue hTLR4 cells (Invitrogen, Waltham, MA) were cultured in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Gaithersburg, MD) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Sigma Aldrich, St. Louis, MO), 100 IU mL−1 penicillin, 100 μg mL−1 streptomycin, 1 mM sodium pyruvate, and 200 mM L-glutamine in a humidified incubator at 37 °C, 5% CO2. For cell stimulation, lyophilized LPS was reconstituted in sterile, endotoxin-free water at a concentration of 1 mg mL−1. This stock was serially diluted in DMEM before addition to the cell culture for the stimulation experiment. Post-stimulation (16 h), supernatants were collected from HEK-Blue cells, and the production of SEAP reporter was detected using Quanti-Blue (Invitrogen) according to the manufacturer’s instructions. NF-κB reporter cell line stimulation data were plotted as the mean (± SD) from biological duplicates using GraphPad Prism 7.0 (La Jolla, CA).
Oyler B.L., Khan M.M., Smith D.F., Harberts E.M., Kilgour D.P., Ernst R.K., Cross A.S, & Goodlett D.R. (2018). Top down tandem mass spectrometric analysis of a chemically modified rough-type lipopolysaccharide vaccine candidate. Journal of the American Society for Mass Spectrometry, 29(6), 1221-1229.
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6

Dual Reporter THP1 Cell Assay

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THP1-Dual™ cells are derived from the human THP-1 monocyte cell line by stable integration of two inducible reporter constructs (Invivogen). As a result, THP1-Dual™ cells allow the simultaneous study of the NF-κB pathway by monitoring the activity of SEAP and the IRF pathway by assessing the activity of a secreted luciferase (Lucia). Both reporter proteins are readily measurable in the cell culture supernatant when using QUANTI-Blue™ (Invitrogen), a SEAP detection reagent, and QUANTI-Luc™ (Invitrogen), a luciferase detection reagent. In brief, supernatants from THP1-stimulated cells were separately incubated with QUANTI-Blue™ or https://www.invivogen.com/quanti-luc according to the manufacturer’s guidelines and read for their luciferase or colorimetric content as a reflection of their IRF and NF-kB activity, respectively.
van der Horst D., Kurmasheva N., Marqvorsen M.H., Assil S., Rahimic A.H., Kollmann C.F., Silva da Costa L., Wu Q., Zhao J., Cesari E., Iversen M.B., Ren F., Jensen T.I., Narita R., Schack V.R., Zhang B.C., Bak R.O., Sette C., Fenton R.A., Mikkelsen J.G., Paludan S.R, & Olagnier D. (2024). SAM68 directs STING signaling to apoptosis in macrophages. Communications Biology, 7, 283.
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