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In April, researchers around the world cited ImmunoChemistry Technologies’ products in their publications. Explore our selection of April citations from 2020.

Zou W, Rohatgi N, Brestoff JR, Moley JR, Li Y, Williams JW, Alippe Y, Pan H, Pietka TA, Mbalaviele G, Newberry EP, Davidson NO, Dey A, Shoghi KI, Head RD, Wickline SA, Randolph GJ, Abumrad NA, Teitelbaum SL. Myeloid-specific Asxl2 deletion limits diet-induced obesity by regulating energy expenditure. J Clin Invest. 2020 May 1;130(5):2644-2656. doi: 10.1172/JCI128687. Full Text

"To study NLRP3 inflammasome activation based on caspase-1 activity, cells were cultured on coverslips and primed with LPS for 4 hours. Cells were stimulated with nigericin for 30 minutes followed by incubation with FLICA FAM-YVAD-FMK probe (ImmunoChemistry Technology LLC) for an additional 30 minutes and analyzed by fluorescence microscopy."

Malhotra S, Costa C, Eixarch H, Keller CW, Amman L, Martínez-Banaclocha H, Midaglia L, Sarró E, Machín-Díaz I, Villar LM, Triviño JC, Oliver-Martos B, Parladé LN, Calvo-Barreiro L, Matesanz F, Vandenbroeck K, Urcelay E, Martínez-Ginés ML, Tejeda-Velarde A, Fissolo N, Castilló J, Sanchez A, Robertson AAB, Clemente D, Prinz M, Pelegrin P, Lünemann JD, Espejo C, Montalban X, Comabella M. NLRP3 inflammasome as prognostic factor and therapeutic target in primary progressive multiple sclerosis patients. Brain. 2020 May 1;143(5):1414-1430. doi: 10.1093/brain/awaa084. Abstract

"Active caspase-1 was measured in monocytes using the specific fluorescent probe FLICA-660 Caspase-1 Assay Kit (Immunochemistry Technologies) following the manufacturer’s instructions. For the determination of ASC specks and FLICA, monocytes were selected using anti-CD14 PE (clone 61D3, Tonbo Biosciences) or anti-CD14 APC-H7 (clone MφP9, BD Biosciences), respectively. Samples were analysed by flow cytometry using a FACS Canto (BD Biosciences) and the FCS express software (De Novo Software)."

Raynor JL, Liu C, Dhungana Y, Guy C, Chapman NM, Shi H, Neale G, Sesaki H, Chi H. Hippo/Mst signaling coordinates cellular quiescence with terminal maturation in iNKT cell development and fate decisions. J Exp Med. 2020 Jun 1;217(6):e20191157. doi: 10.1084/jem.20191157. Abstract

"For staining mitochondria, intracellular ROS, and 2-NBDG uptake, unfixed lymphocytes were incubated for 30 min at 37°C with 10 nM MitoTracker Green or MitoTracker Deep Red (Life Technologies, M7514 and MM22426), 10 nM TMRM (ImmunoChemistry Technologies, 9105), or 2.5 µM CellROX Deep Red (Life Technologies, C10422), or for 1 h at 37°C with 30 µM 2-NBDG (Thermo Fisher Scientific, N13195) before staining surface markers. For assessment of mitochondria and cellular ROS in fixed cells, MitoTracker Deep Red and CellROX Deep Red were used per the..."

Wang X, Chang CH, Jiang J, Liu X, Li J, Liu Q, Liao YP, Li L, Nel AE, Xia T. Mechanistic Differences in Cell Death Responses to Metal-Based Engineered Nanomaterials in Kupffer Cells and Hepatocytes. Small. 2020 May;16(21):e2000528. doi: 10.1002/smll.202000528. Epub 2020 Apr 26. Abstract

"A particle library of 16 materials was obtained from The NIEHS NHIR Consortium. The CellTiter 96 AQueous MTS assay and GSH‐Glo glutathione assay kits were purchased from Promega (Madison, WI). The FAM‐FLICA Caspase 1, Caspase 3/7, and Magic Red Cathepsin B assay kits were purchased from ImmunoChemistry Technologies, LLC (Bloomington, MN). The lipopolysaccharide (LPS) and GSDMD, NLRP3, and caspase 1 siRNAs were obtained from Sigma (St. Louis, MO). The ELISA kit for mouse IL‐1β was purchased from R&D Systems (Minneapolis, MN). The ELISA kit for the mouse IL‐1β pro‐form was purchased from Thermo Fisher Scientific (Waltham, MA). The mouse Kupffer line, KUP5, was purchased from Riken..."

Elsherbini A, Kirov AS, Dinkins MB, Wang G, Qin H, Zhu Z, Tripathi P, Crivelli SM, Bieberich E. Association of Aβ with ceramide-enriched astrosomes mediates Aβ neurotoxicity. Acta Neuropathol Commun. 2020 Apr 28;8(1):60. doi: 10.1186/s40478-020-00931-8. Full Text

"The FLICA 660 Poly Caspase Assay Kit (ImmunoChemistry Technologies, Minnesota, USA) was used to determine the presence of early caspase activation. This in vitro assay employs the fluorescent inhibitor probe 660-VAD-FMK to label active caspase enzymes in living cells. N2a cells (0.25–1·105) were incubated with exosomes (0.5–1·104 exosomes/cell) for 6 h at 37 °C. The cells were washed twice with PBS and resuspended in RPMI medium with 10% FBS before staining with 30 × FAM-VAD-FMK for 30 min at 37 °C. Cells were washed with 1 x..."

Suschak JJ, Dupuy LC, Shoemaker CJ, Six C, Kwilas SA, Spik KW, Williams JA, Schmaljohn CS. Nanoplasmid Vectors Co-Expressing Innate Immune Agonists Enhance DNA Vaccines for Venezuelan Equine Encephalitis Virus and Ebola Virus. Mol Ther Methods Clin Dev. 2020 Apr 15;17:810-821. doi: 10.1016/j.omtm.2020.04.009. Online ahead of print. Full Text

"The following day, plates were washed with PBS containing 0.05% Tween-20 and then blocked with Neptune Block (ImmunoChemistry Technologies, Bloomington, MN) for 2 h at 37°C. Plates were washed again prior to being loaded with 2-fold serial dilutions of mouse sera in duplicate (dilution range 1:200 to 1:25,600). Serum dilutions were carried out in Neptune Block. Plates were incubated at ambient temperature for 1 h prior to being washed, and then incubated with a 1:1,000 dilution of horseradish peroxidase (HRP) conjugated goat anti-mouse (SeraCare Life Sciences, Gaithersburg, MD) in Neptune Block for 1 h at ambient temperature."

Kaproń B, Czarnomysy R, Wysokiński M, Andrys R, Musilek K, Angeli A, Supuran CT, Plech T. 1,2,4-Triazole-based anticonvulsant agents with additional ROS scavenging activity are effective in a model of pharmacoresistant epilepsy. J Enzyme Inhib Med Chem. 2020 Dec;35(1):993-1002. doi: 10.1080/14756366.2020.1748026. Full Text

"Total ROS activity was measured using Intracellular Total ROS Activity Assay Kit (ImmunoChemistry Technologies, Bloomington, MN, USA). After 24 h of incubation of glioblastoma cells U-87 MG (ATCC, Manassas, VA, USA) with the test compounds in concentration 10 µg/ml, the medium was removed, the cells were washed twice with the cold PBS solution and the Assay Buffer (provided by the kit manufacturer) was added. Then, cells (at a density of 1 × 106 cells/ml) were treated with 10 µl of Total ROS green reagent and incubated for 1 h at 37 °C in a CO2 incubator. After incubation, the cells were subjected to a rinsing procedure using the Assay Buffer."

Products Used:
FAM-FLICA Caspase-1 Assay
FLICA 660 Caspase-1 Assay
MitoPT TMRM Assay
FAM-FLICA Caspase-3/7 Assay
Magic Red Cathepsin B Assay
FLICA 660 Poly Caspase Assay
Neptune Block ELISA Blocking Buffer
Intracellular Total ROS Activity Assay