Alexa fluor 647 conjugated goat anti mouse fab

Manufactured by Jackson ImmunoResearch

Alexa-Fluor-647–conjugated goat anti-mouse Fab is a secondary antibody that binds to the Fab region of mouse primary antibodies. The antibody is conjugated to Alexa Fluor 647, a fluorescent dye with excitation and emission maxima at 650 nm and 665 nm, respectively.

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Lab products found in correlation

Facsaria cell sorter, by BD (2 mentions) Buv 395cd4, by BD (1 mentions) Annexin 5 percp cy5, by BioLegend (1 mentions) Countbright beads, by Thermo Fisher Scientific (1 mentions) Cd8 apc cy7, by BD (1 mentions) Zombie nir, by BioLegend (1 mentions) Lsr 2, by BD (1 mentions) Foxp3 transcription factor staining kit, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Alexa Fluor 647 (152 citations) Alexa 647 (67 citations) Alexa Fluor 647 anti-mouse (12 citations) Anti goat Alexa Fluor 647 (3 citations)

2 protocols using alexa fluor 647 conjugated goat anti mouse fab

Comprehensive Cytometric Analysis of CAR T Cells

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All antibodies were titrated. CAR expression was measured with Alexa-Fluor-647–conjugated goat anti-mouse Fab (Jackson ImmunoResearch cat#115-607-003, RRID: AB_2338931). The following antibodies were used: CD19–PE (clone SJ25C1 BD Biosciences cat# 340364, RRID: 400018) or CD19–BUV395 (clone SJ25C1, BD Biosciences cat# 563551, RRID: AB_2738272), CD19–BV510 (clone SJ25C1, Biolegend cat#363020, RRID: AB_2564229), CD3–BUV737 (clone UCHT, BD Biosciences cat# 612750, RRID: RRID:AB_2870081), BUV-395CD4 (BD Biosciences cat# 563552, RRID:AB_2738273), and APC-cy7-CD8 (BD Biosciences cat# 557834, RRID:AB_396892). CountBright beads (Invitrogen cat# C36950) were used to determine the absolute number of cells according to the manufacturer’s protocol. 7-AAD or DAPI was used to exclude dead cells. Fc Receptor–Binding Inhibitor Antibody Human (eBioscience cat# 14-9161-73, RRID: AB_468582) was used to block Fc receptors. In vitro viability after RT was assessed using Zombie NIR (Biolegend cat# 423106) and Annexin V PerCP-Cy5.5 (Biolegend cat# 640936) according to the manufacturer’s protocols. Data were collected using BD LSR-II, BD LSR-Fortessa, and Cytek Northern Lights cytometers. Data were analyzed with FlowJo Software (Treestar/BD Biosciences, RRID: SCR_008520). Cell sorting was performed using a BD FACSAria cell sorter.

Kim A.B., Chou S.Y., Kang S., Kwon E., Inkman M., Szymanski J., Andruska N., Colgan C., Zhang J., Yang J.C., Singh N, & DeSelm C.J. (2023). Intrinsic tumor resistance to CAR T cells is a dynamic transcriptional state that is exploitable with low-dose radiation. Blood Advances, 7(18), 5396-5408.

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Cell Surface and Intracellular Phenotyping

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CAR expression was measured with Alexa-Fluor-647-conjugated goat anti-mouse Fab (Jackson ImmunoResearch, 115-606-072). Flow cytometry antibodies used for cell surface phenotyping are provided in Supplementary Table 1. For intracellular staining, cells were fixed and permeabilized using Foxp3/Transcription Factor staining kit (eBioscience, 00-5523-00) according to manufacturer’s protocol. Flow cytometry antibodies used for intra-cellular studies are provided in Supplementary Table 5. Data was analysed by FlowJo v10.1 (BD). Cell sorting was performed on a BD FACSAria cell sorter. Gating strategies for flow cytometry are provided in SI Figure 4.

Jain N., Zhao Z., Feucht J., Koche R., Iyer A., Dobrin A., Mansilla-Soto J., Yang J., Zhan Y., Lopez M., Gunset G, & Sadelain M. (2023). TET2 guards against unchecked BATF3-induced CAR T cell expansion. Nature, 615(7951), 315-322.

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