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S7575

Manufactured by Thermo Fisher Scientific
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The S7575 is a laboratory equipment product manufactured by Thermo Fisher Scientific. It is designed to perform specific functions within a laboratory setting. The core function of this product is to [provide detailed description of the core function without extrapolation].

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Lab products found in correlation

Ab196147, by Novus Biologicals (2 mentions) Mabn92 af647, by Abcam (2 mentions) Nbp2 33184af532, by Thermo Fisher Scientific (2 mentions) Syto13, by Thermo Fisher Scientific (1 mentions) Bond rx, by Leica (1 mentions) Bond rx ffpe, by Leica (1 mentions) Cd68 647, by Novus Biologicals (1 mentions) Cd45 594, by NanoString (1 mentions) Panck 532, by NanoString (1 mentions) Lipofectamine 3000 transfection reagent, by Thermo Fisher Scientific (1 mentions) Bond platform, by Leica (1 mentions) Buffer w, by NanoString (1 mentions)

4 protocols using s7575

1

GeoMx DSP Tissue Slide Preparation

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For GeoMx DSP slide preparation, we followed the GeoMx DSP slide prep user manual (MAN-10087-04). Briefly, tissue slides were baked in a drying oven at 60 °C for 1 hour and then loaded to Leica Biosystems BOND RX FFPE for deparaffinization and rehydration. After the target retrieval step, tissues were treated with Proteinase K solution to expose RNA targets followed by fixation with 10% NBF. After all tissue pre-treatments were done, tissue slides were unloaded from the Leica Biosystems BOND RX and incubated with RNA probe mix (COVID-19 Immune Response Atlas panel) overnight. The next day, tissues were washed and stained with tissue visualization markers; CD68-647 at 1:400 (Novus Bio, NBP2-34736AF647), CD45-594 at 1:10 (NanoString Technologies), PanCK-532 at 1:20 (NanoString Technologies) and/or SYTO 13 at 1:10 (Thermo Scientific S7575).
Park J., Foox J., Hether T., Danko D.C., Warren S., Kim Y., Reeves J., Butler D.J., Mozsary C., Rosiene J., Shaiber A., Afshin E.E., MacKay M., Rendeiro A.F., Bram Y., Chandar V., Geiger H., Craney A., Velu P., Melnick A.M., Hajirasouliha I., Beheshti A., Taylor D., Saravia-Butler A., Singh U., Wurtele E.S., Schisler J., Fennessey S., Corvelo A., Zody M.C., Germer S., Salvatore S., Levy S., Wu S., Tatonetti N.P., Shapira S., Salvatore M., Westblade L.F., Cushing M., Rennert H., Kriegel A.J., Elemento O., Imielinski M., Rice C.M., Borczuk A.C., Meydan C., Schwartz R.E, & Mason C.E. (2022). System-wide transcriptome damage and tissue identity loss in COVID-19 patients. Cell Reports Medicine, 3(2), 100522.
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2

Transfection of HCC Cell Lines

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Human hepatocellular carcinoma cell lines Hep3B, Huh‐7, PLC/PRF/5, SNU449, SNU368, and SNU475 cells (detailed information on cell lines can be found at https://cellbank.snu.ac.kr/main/index.html) were certified through the short tandem repeat genotyping and were acquired in March 2021 from the Korean Cell Line Bank (Seoul, South Korea) and cultured in DMEM or RPMI‐1640 containing 10% FBS and 100 units/mL penicillin‐streptomycin. The HCC cell line used in all in vivo assays was Huh‐7. Cells were transiently transfected with small interfering RNA (siRNAs) against ITGB1 (s7575, Thermo Scientific, Waltham, MA, USA), ITGB5 (s7590, Thermo Scientific), or negative control (4457287, Thermo Scientific) using Lipofectamine3000 transfection reagent (Invitrogen), according to the manufacturer's instructions.
Eun J.W., Yoon J.H., Ahn H.R., Kim S., Kim Y.B., Lim S.B., Park W., Kang T.W., Baek G.O., Yoon M.G., Son J.A., Weon J.H., Kim S.S., Cho H.J, & Cheong J.Y. (2023). Cancer‐associated fibroblast‐derived secreted phosphoprotein 1 contributes to resistance of hepatocellular carcinoma to sorafenib and lenvatinib. Cancer Communications, 43(4), 455-479.
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3

GeoMx DSP Slide Preparation Protocol

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Slides were prepared by baking in a drying oven and then processed using the Leica BOND platform (Leica Biosystems) as specified by the NanoString GeoMx DSP Slide Preparation User Manual. Briefly, slides were deparaffinized with xylene then rehydrated through a graded ethanol series. Targets were exposed by incubating the slides in a pressure cooker at 100°C in 1X Tris-EDTA buffer, pH9 followed by proteinase K digestion. Next, the GeoMx NGS-RNA Probe Mix (NanoString Technologies) was applied to each slide, and slides were incubated at 37°C for 16-24 hours. This probe mix contained barcoded oligonucleotide probes against 1,700 gene targets and included internal positive and negative control probes. After washing, the slides were then blocked in Buffer W (NanoString Technologies) and stained with morphology markers for 1 hour. The fluorescently conjugated morphology markers used are as follows: Iba1 at 1:75 dilution (EMD Millipore, MABN92-AF647), CD3 at 1:50 dilution (Abcam, ab196147), GFAP at 1:3000 dilution (Novus Biologicals, NBP2-33184AF532), and SYTO 13 nuclei acid stain at 1:5 dilution (Thermo Scientific S7575).
Kim Y., Danaher P., Cimino P.J., Hurth K., Warren S., Glod J., Beechem J.M., Zada G, & McEachron T.A. (2023). Highly multiplexed spatially resolved proteomic and transcriptional profiling of the glioblastoma microenvironment using archived formalin-fixed paraffin-embedded specimens.. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, 36(1), 100034.
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4

Multimodal Immunohistochemistry Protocol

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Slides were baked in a drying oven, deparaffinized, rehydrated in an ethanol gradient, and placed in ditheyl pyrocarbonate (DEPC)-treated water. Antigen retrieval was performed with 1X citrate buffer,pH6 in a pressure cooker on high temperature setting for 15 minutes. The slides were then blocked and incubated with the primary antibody mix and the fluorescently conjugated morphology markers. The morphology markers used are as follows: Iba1-647 at 1:75 dilution (EMD Millipore, MABN92-AF647), CD3-594 at 1:50 dilution (Abcam, ab196147), GFAP-532 at 1:3000 dilution (Novus Biologicals, NBP2-33184AF532), and SYTO 13 at 1:5 dilution (Thermo Scientific S7575).
Kim Y., Danaher P., Cimino P.J., Hurth K., Warren S., Glod J., Beechem J.M., Zada G, & McEachron T.A. (2023). Highly multiplexed spatially resolved proteomic and transcriptional profiling of the glioblastoma microenvironment using archived formalin-fixed paraffin-embedded specimens.. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, 36(1), 100034.
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