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

Crespo C, Mulik S, Dotiwala F, Ansara JA, Sen Santara S, Ingersoll K, Ovies C, Junqueira C, Tilburgs T, Strominger JL, Lieberman J. Decidual NK Cells Transfer Granulysin to Selectively Kill Bacteria in Trophoblasts. Cell. 2020 Sep 3;182(5):1125-1139.e18. doi: 10.1016/j.cell.2020.07.019. Epub 2020 Aug 20. Abstract 

"BSA Sigma Cat.# A9418-500G CAS: 9048-46-8 Triton X-100 Sigma Cat.# X100-500ML; CAS: 9002-93-1 Hoechst 33342 Immunochemistry Technologies Cat.# 639 DAPI Thermo Scientific Cat.# 62248..."

Gaul S, Leszczynska A, Alegre F, Kaufmann B, Johnson CD, Adams LA, Wree A, Damm G, Seehofer D, Calvente CJ, Povero D, Kisseleva T, Eguchi A, McGeough MD, Hoffman HM, Pelegrin P, Laufs U, Feldstein AE. Hepatocyte pyroptosis and release of inflammasome particles induce stellate cell activation and liver fibrosis. J Hepatol. 2020 Aug 4;S0168-8278(20)30522-5. doi: 10.1016/j.jhep.2020.07.041. Online ahead of print. Abstract 

"Liver tissue was collected and stained for α-SMA, TUNEL positive cells and picrosirius red as previously described <20>. To detect caspase-1 activity in frozen liver tissue slides, we used the FAM-Flica caspase-1 Assay Kit (ImmunoChemistry Technologies, Bloomington, MN, USA) according to manufacturer’s instructions. Flica-positive cells were quantified by ImageJ and normalized to the total number of DAPI-stained cells. F4/80 and Cd11b staining was performed as described previously <21>. Positive cells were counted in 10X magnification images and normalized to DAPI stained cells. Immunofluorescent staining of α-SMA was performed on acetone fixed cells and relative fluorescence intensity was normalized on total cell count."

Gioulbasani M, Galaras A, Grammenoudi S, Moulos P, Dent AL, Sigvardsson M, Hatzis P, Kee BL, Verykokakis M. The transcription factor BCL-6 controls early development of innate-like T cells. Nat Immunol. 2020 Sep;21(9):1058-1069. doi: 10.1038/s41590-020-0737-y. Epub 2020 Jul 27. Abstract 

"Thymocyte suspensions from 10-day-old or 3–6-week-old mice were incubated with anti-FcγR before staining with fluorochrome-conjugated antibodies. Cells were acquired in a FACSCanto II or LSRII Fortessa or sorted in a FACSAria III and analyzed with FlowJo. Enrichment of thymocytes for iNKT cells was performed by staining total thymocytes with APC- or PE-conjugated CD1dPBS57 tetramers (National Institutes of Health (NIH) Tetramer Facility at Emory University), followed by anti-APC or anti-PE microbeads and subjected to MACS-based magnetic cell separation. Similarly, enrichment of thymocytes for MAIT cells was performed by staining with APC-conjugated MR1-OP-5RU tetramer (NIH tetramer facility at Emory University) for 1h at 25 ° C. Propidium iodide was included in all samples to exclude dead cells from the analysis. Pan-caspase staining was performed with the carboxyfluorescein (FAM)-FLICA in vitro Caspase Detection kit (ImmunoChemistry Technologies), according to the manufacturer’s instructions. To calculate ST0 cell numbers, a small fraction of the thymus was stained with Tetramer, TCRβ and CD24 before tetramer enrichment and analyzed by flow cytometry. From this staining, the total number of iNKT cells was calculated, as Tetr+TCRβ+ cells. Then, the number of ST0–ST3 cells was calculated according to the percentages in enriched samples. Antibodies specific for the following antigens were purchased from BD Biosciences, eBiosciences, BioLegend and Cell Signaling."

Jäger E, Murthy S, Schmidt C, Hahn M, Strobel S, Peters A, Stäubert C, Sungur P, Venus T, Geisler M, Radusheva V, Raps S, Rothe K, Scholz R, Jung S, Wagner S, Pierer M, Seifert O, Chang W, Estrela-Lopis I, Raulien N, Krohn K, Sträter N, Hoeppener S, Schöneberg T, Rossol M, Wagner U. Calcium-sensing receptor-mediated NLRP3 inflammasome response to calciprotein particles drives inflammation in rheumatoid arthritis. Nat Commun. 2020 Aug 25;11(1):4243. doi: 10.1038/s41467-020-17749-6. Full Text 

"DETECTION OF CATHEPSIN ACTIVITY Magic Red Cathepsin-B Assay (ImmunoChemistry Technologies) was used for the detection of Cathepsin B activity in monocytes. Assay was performed as described in manufacturer’s instructions. Cathepsin activity was determined after 3 h of incubation in either 1 or 5.6 mM -containing cell culture medium. Fluorescence was measured with a Tecan infinite M200 plate reader (Ex 592 nm/ Em 628 nm) and Tecan Magellan V7.2."

Ma L, Wang L, Nelson AT, Han C, He S, Henn MA, Menon K, Chen JJ, Baek AE, Vardanyan A, Shahoei SH, Park S, Shapiro DJ, Nanjappa SG, Nelson ER. 27-Hydroxycholesterol acts on myeloid immune cells to induce T cell dysfunction, promoting breast cancer progression. Cancer Lett. 2020 Aug 28;493:266-283. doi: 10.1016/j.canlet.2020.08.020. Online ahead of print. Abstract 

"...harvested 24, 48 or 72 h s after co-culture for apoptosis assay using FAM-DEVD-FMK (FAM-FLICA) against active caspase-3/7 and propidium iodide (ImmunoChemistry Technologies). Naïve CD4+ T cells were isolated from wildtype mice, chemically activated and cultured in conditioned media from treated BMDMs, harvested after 24 or 48 h s and assayed for apoptosis (FAM-FLICA + PI) as described above." Unbehau R, Luthringer-Feyerabend B, Willumeit-Römer R. The impact of brain cell metabolism and extracellular matrix on magnesium degradation. Acta Biomaterialia. 2020 Sep 2;S1742-7061(20)30513-4. doi: 10.1016/j.actbio.2020.08.043. Online ahead of print. Full Text 

"ECM distribution was analyzed via staining of GAGs with the fluorescein isothiocyanate (FITC) conjugated lectins(Vector Laboratories Inc., Burlingame, USA) wheat germ agglutinin (WGA) and soybean agglutinin (SBA), which specifically bind to sugar moieties of GAGs. Further, Col-F collagen binding reagent (ImmunoChemistry TechnologiesLLC, Bloomington, USA), which shows an unspecific affinity to collagen and elastin, was used to visualize collagenous parts of the ECM."

Sun X, Shu Y, Yan P, Huang H, Gao R, Xu M, Lu L, Tian J, Huang D, Zhang J. Transcriptome profiling analysis reveals that ATP6V0E2 is involved in the lysosomal activation by anlotinib. Cell Death Dis. 2020 Aug 24;11(8):702. doi: 10.1038/s41419-020-02904-0. Full Text 

"Lysosomal function was also estimated by the cathepsin B and L enzymatic activity. After designated treatment, cells were further loaded with Magic RedTM cathepsin B (Immunochemistry Technologies, 938) or cathepsin L (Immunochemistry Technologies, 942) reagents for 30 min. The cells were collected and the fluorescence intensities of 10,000 cells per sample were measured by flow cytometry. We recorded the fluorescence of Magic Red using the FL-2 channel of FACs (BD Biosciences)." 

Products Used: 
Hoechst 33342 Fluorescent Nucleic Acid Stain 
FAM-FLICA Caspase-1 Assay 
FAM-FLICA Poly Caspase Assay 
Magic Red Cathepsin B Assay 
FAM-FLICA Caspase-3/7 Assay 
Col-F Collagen Binding Reagent 
Magic Red Cathepsin L Assay