4e3d10h2 e3

Manufactured by Thermo Fisher Scientific

Sourced in United States

The 4E3D10H2/E3 is a laboratory instrument designed for analytical purposes. It features advanced functional capabilities to support various scientific applications. The device specifications and core functions are presented in a factual and objective manner without interpretation or extrapolation.

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Lab products found in correlation

Rpa t8, by BioLegend (1 mentions) Celltrace violet, by Thermo Fisher Scientific (1 mentions) Streptavidin apc, by Thermo Fisher Scientific (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Human fc block, by BD (1 mentions) Eh12.2h7, by BioLegend (1 mentions) Rpa t4, by BioLegend (1 mentions) Sc 374171, by Santa Cruz Biotechnology (1 mentions) Hrp coupled secondary antibody, by Jackson ImmunoResearch (1 mentions) Immobilon forte, by Merck Group (1 mentions)

3 protocols using 4e3d10h2 e3

Flow Cytometry Analysis of CAR T Cells

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To detect CD19 CAR expression, biotinylated CD19 (10 μg ml⁻¹; Acro Biosystems CD9-H8259) and streptavidin-APC (ThermoFisher S868) were used according to the manufacturer’s instructions. NKG2DL expression was assessed by staining with NKG2D-Fc chimera (10 μg ml⁻¹; Fisher 1299NK050) followed by an αFc secondary stain (Invitrogen A-10631). NIR Live/Dead (ThermoFisher L34976), CFSE (LifeTech C34554) and CellTrace Violet (LifeTech C34557) were used according to the manufacturer’s instructions. Human Fc block (BD 564220) was used before staining with any antibodies. Antibodies for CD69 (FN50; BD, RRID: AB_1727510), CD4 (RPA-T4; BioLegend, RRID: AB_314076); hCD8 (RPA-T8; BioLegend, RRID: AB_10669564), CD3 (UCHT1; BD, RRID: AB_2744387), CD45 (HI30; BD, RRID: AB_395875), CD19 (HIB19; BioLegend, RRID: AB_395813), PD-1 (EH12.2H7; Biolegend, RRID: AB_10900818), CD107a (H4A3; Biolegend, RRID: AB_2562647), mCD8 (53-6.7; BioLegend, RRID: AB_10897101), HER2 (24D2; Biolegend, RRID: AB_756121), and HisTags (4E3D10H2/E3; ThermoFisher, RRID: AB_2610637) and human Fc (HP6069; ThermoFisher, RRID: AB_2534050) were all used at 1:100 dilutions.

Miller I.C., Zamat A., Sun L.K., Phuengkham H., Harris A.M., Gamboa L., Yang J., Murad J.P., Priceman S.J, & Kwong G.A. (2021). Enhanced intratumoural activity of CAR T cells engineered to produce immunomodulators under photothermal control. Nature biomedical engineering, 5(11), 1348-1359.

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Binding Assay for VE-PTP and GEF-H1

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To confirm direct binding between VE-PTP and GEF-H1, a binding assay was performed. Briefly, binding buffers containing 20 mM Tris-HCl, pH 7.5, 100 mM, 1 mM mercaptoethanol, and 1% Triton X-100 (Lansbergen et al., 2004 (link)) were used for binding assay experiments. 10 µg GST-tagged GEF-H1 and 10 µg His-tagged VE-PTP purified proteins were incubated with binding buffer at 4°C for 90 min. His-tagged VE-PTP was pulled down using HisPur Ni-NTA beads, run on SDS-PAGE, and probed with GST (anti-mouse; sc-374171; Santa Cruz) or His antibody (anti-mouse; 4E3D10H2/E3; ThermoFisher).

Juettner V.V., Kruse K., Dan A., Vu V.H., Khan Y., Le J., Leckband D., Komarova Y, & Malik A.B. (2019). VE-PTP stabilizes VE-cadherin junctions and the endothelial barrier via a phosphatase-independent mechanism. The Journal of Cell Biology, 218(5), 1725-1742.

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SDS-PAGE and Western Blot Analyses

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Tris/glycine and taurine/imidazole SDS-PAGE and Western blots were performed as described previously [38 (link)]. Bcl-xL was revealed with rabbit monoclonal antibody E18 (Abcam), His₇-tag was revealed with mouse monoclonal antibody 4E3D10H2/E3 (Thermofisher), and Bax was revealed with mouse monoclonal antibody 2D2 (Santa-Cruz, Santa Cruz, CA, USA). HRP-coupled secondary antibodies were from Jackson Laboratories, and blots were revealed with Immobilon Forte (Millipore, Burlington, MA, USA). Images were captured with a G-Box digital camera (Syngene, Cambridge, UK) and processed with Image J 1.48v (https://imagej.nih.gov/ij/index.html, downloaded 24 May 2019).

Rouchidane Eyitayo A., Boudier-Lemosquet A., Chaignepain S., Priault M, & Manon S. (2023). Bcl-xL Is Spontaneously Inserted into Preassembled Nanodiscs and Stimulates Bax Insertion in a Cell-Free Protein Synthesis System. Biomolecules, 13(6), 876.

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