Gallios b43618

Manufactured by Beckman Coulter

Sourced in United States

The Gallios B43618 is a flow cytometer designed for research applications. It is capable of multi-color analysis and can detect a wide range of fluorescent parameters. The Gallios B43618 provides high-performance data acquisition and analysis capabilities to support various research projects.

Automatically generated - may contain errors

Lab products found in correlation

Attune nxt, by Thermo Fisher Scientific (4 mentions) 123count ebeads, by Thermo Fisher Scientific (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Rm4 5, by Thermo Fisher Scientific (1 mentions) Ebio1d3, by Thermo Fisher Scientific (1 mentions) C9b7w, by Thermo Fisher Scientific (1 mentions) Facsaria, by BD (1 mentions) 7 aminoactinomycin d, by BioLegend (1 mentions) Dapi exclusion, by Thermo Fisher Scientific (1 mentions)

4 protocols using gallios b43618

Multicolor Flow Cytometry Analysis of CAR T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

10-color Gallios B43618 (Beckman Coulter, Indianapolis, IN) and 14-color Attune NxT (ThermoFisher Scientific) were used to acquire data. Analysis was performed with FlowJo software. Cells were counted with 123count eBeads (Thermo Fisher). Expression of CAR was detected by myc tag (9E10, Alexa Fluor 647, ThermoFisher) or violet-excitable GFP tag. DAPI (0.5 mg/ml, Sigma-Aldrich) or a LIVE/DEAD fixable yellow fluorescent dye (Thermo Fisher) were used to exclude dead cells in all experiments. Sorting of splenocytes after tissue processing was done using a BD FACSAria under sterile conditions. Antibodes to the following mouse proteins were used for flow cytometry: CD3ε (145-2C11,eBioscience, cat# 11-0031-85), CD3 (17A2, BioLegend, cat# 100233), CD4 (GK1.5, eBioscience, cat# 48-0041-80), CD4 (RM4-5, eBioscience, cat# 69-0042-80), CD8α (53-6.7, eBioscience, cat# 48-0041-80), CD19 (eBio1D3, eBioscience, cat# 48-0193-82), CD25 (PC61.5, eBioscience, cat# 25-0251-81), CD45 (30-F11, BioLegend, cat# 103106), CD80 (16-10A1, eBioscience, cat# 46-0801), CD86 (GL1, eBioscience, cat# 25-0862), CD223LAG-3 (C9B7W, eBioscience, cat# 12-5956-80), CD279 PD-1 (J43, eBiosceince, cat# 48-9985-82), and TIM3(8B.2C12, eBiosceince, cat# 48-5871-80).

Wijewarnasuriya D., Bebernitz C., Lopez A.V., Rafiq S, & Brentjens R. (2020). Excessive costimulation leads to dysfunction of adoptively transferred T cells. Cancer immunology research, 8(6), 732-742.

+ Open protocol

+ Expand

Flow Cytometric Analysis of Transduced Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Flow cytometric analyses were performed using 10-color Gallios B43618 (Beckman Coulter, Indianapolis, Indiana, USA) and 14-color Attune NxT (Thermo Fisher Scientific) instruments. Data were analyzed using FlowJo (Tree Star) V.10.8.0. Flow cytometry was used to determine transduction efficiency of transduced cells following staining with phycoerythrin (PE)-conjugated cetuximab antibody and Myc-tag (9B11, Alexa Fluor 647, Cell Signaling). DAPI (0.5 mg/mL, Sigma Aldrich) or 7-amino-actinomycin D (0.05 mg/mL, BioLegend) and PO-PRO-1 Iodine (Invitrogen) staining were used to exclude dead cells in all experiments. Antibodies are listed in online supplemental table 1. All antibodies were purchased from BioLegend, BD Biosciences, Invitrogen or eBioscience.

Freeman R., Shahid S., Khan A.G., Mathew S.C., Souness S., Burns E.R., Um J.S., Tanaka K., Cai W., Yoo S., Dunbar A., Park Y., McAvoy D., Hosszu K.K., Levine R.L., Boelens J.J., Lorenz I.C., Brentjens R.J, & Daniyan A.F. (2024). Developing a membrane-proximal CD33-targeting CAR T cell. Journal for Immunotherapy of Cancer, 12(5), e009013.

+ Open protocol

+ Expand

Flow Cytometry Analysis of CAR Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

A 10-color Gallios B43618 (Beckman Coulter) was used to acquire data. Analysis was performed with FlowJo software (V10, Tree Star). Expression of CAR was determined by surface staining using either BCMA ECD-Fc (shared by Eureka Therapeutics), GPRC5D-scFv anti-idiotype (generated by Charles River Labs and shared by Bristol-Myers Squibb), or anti-human IgG4 antibody to the shared spacer region (clone EP4420; Abcam). Anti-IgG4 primary was conjugated with lightning link labeling kits (Innova Biosciences, Novus Biologicals). BCMA antigen detected using clone FAB193A (R&D systems). Cells were counted with 123count eBeads (ThermoFisher). Viability was determined by DAPI exclusion (ThermoFisher) and cells were gated for low DAPI before further analysis.

Fernández de Larrea C., Staehr M., Lopez A.V., Ng K.Y., Chen Y., Godfrey W.D., Purdon T.J., Ponomarev V., Wendel H.G., Brentjens R.J, & Smith E.L. (2020). Defining an Optimal Dual-Targeted CAR T-cell Therapy Approach Simultaneously Targeting BCMA and GPRC5D to Prevent BCMA Escape–Driven Relapse in Multiple Myeloma. Blood Cancer Discovery, 1(2), 146-154.

+ Open protocol

+ Expand

EPHB2 Expression Dynamics in Induced Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

After 48 hours of Dox treatment, the culture medium of the induced cells was replaced with fresh medium and cells were kept in culture for 10 days [after 48-hour induction, halting Dox treatment at D0 (Day 0) and verifying GFP expression at D2, D5, and D10]. To assess EPHB2 levels, cells were labeled with APC (AlloPhycoCyanin) Mouse Anti-Human EPHB2 (BD Pharmingen). APC Mouse Immunoglobulin G1 (IgG1), K isotype antibody was used for control cell labeling. Calcein violet 450 acetoxymethyl (AM) (Thermo Fisher Scientific) labeling dye was used to detect live cells. The fluorescence intensity of GFP, violet AM, and APC was acquired by a Gallios (B43618, Beckman Coulter) cytometer with FL1, FL9, and FL6.

Keskin T., Rucci B., Cornaz-Buros S., Martin P., Fusco C., Broye L., Cisarova K., Perez E.M., Letovanec I., La Rosa S., Cherix S., Diezi M., Renella R., Provero P., Suvà M.L., Stamenkovic I, & Riggi N. (2021). A live single-cell reporter assay links intratumor heterogeneity to metastatic proclivity in Ewing sarcoma. Science Advances, 7(27), eabf9394.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!