Fluor 647

Manufactured by Thermo Fisher Scientific

Fluor 647 is a fluorescent dye used for labeling and detection applications in biological research. It has an excitation maximum at 650 nm and an emission maximum at 665 nm, making it suitable for detection in the red/far-red region of the visible spectrum.

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Lab products found in correlation

Rabbit iba1 antibody, by Fujifilm (2 mentions) 880 inverted confocal airy scan, by Zeiss (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) P1379, by Merck Group (2 mentions) Anti grp94, by Enzo Life Sciences (1 mentions) Anti 4e bp1, by Cell Signaling Technology (1 mentions) Anti phospho gcn2 t899, by Abcam (1 mentions) Goat anti rat igg, by Thermo Fisher Scientific (1 mentions) Anti gcn2, by Cell Signaling Technology (1 mentions) Anti phospho rps6, by Cell Signaling Technology (1 mentions) Goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Anti phospho 4ebp1, by Cell Signaling Technology (1 mentions) Anti pkr, by Cell Signaling Technology (1 mentions) Alexa fluor 647, by Thermo Fisher Scientific (1 mentions) Anti p erk, by Cell Signaling Technology (1 mentions) Goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594, by Thermo Fisher Scientific (1 mentions) Anti eif2α, by Cell Signaling Technology (1 mentions) Anti gapdh, by Proteintech (1 mentions) Goat anti mouse igg, by Jackson ImmunoResearch (1 mentions)

3 protocols using fluor 647

Antibody Immunoblotting and Immunofluorescence

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Primary antibodies used were anti-viperin (MaP.VIP) (Wang et al., 2007 (link)), anti-GRP94 (Enzo), anti-GAPDH (Proteintech), anti-phospho-4EBP1 (Cell Signaling), anti-4EBP1 (Cell Signaling), anti-PRS6 (Cell Signaling), anti-phospho-RPS6 (Cell Signaling; 2217), anti-phospho-eIF2α (S51) (Cell Signaling), anti-eIF2α (Cell Signaling), anti-Flaviviral E protein (Millipore), anti-GCN2 (Cell Signaling), anti-phospho-GCN2 (T899) (Abcam), anti-PERK (Cell Signaling), anti-HRI (Proteintech), anti-PKR (Cell Signaling), anti-ZAK (Bethyl). All secondary antibodies used for immunofluorescence imaging (goat antimouse IgG coupled to Alexa Fluor 488, Alexa Fluor 647, goat anti-rat IgG coupled to Alexa Fluor 546, Alexa Fluor 594 and goat anti-rabbit IgG coupled to Alexa Fluor 647) were purchased from Invitrogen, and those for immunoblotting (goat anti-mouse IgG and anti-rabbit and anti-rat IgG coupled to horseradish peroxidase and alkaline phosphatase) were purchased from Jackson ImmunoResearch.

Hsu J.C., Laurent-Rolle M., Pawlak J.B., Xia H., Kunte A., Hee J.S., Lim J., Harris L.D., Wood J.M., Evans G.B., Shi P.Y., Grove T.L., Almo S.C, & Cresswell P. (2022). Viperin triggers ribosome collision-dependent translation inhibition to restrict viral replication. Molecular cell, 82(9), 1631-1642.e6.

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Microglial Morphology Analysis via Immunostaining

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Immunostaining and analyses of microglial morphology were performed on the PFA-perfused slices using ionizing calcium-binding adaptor molecule 1 (Iba1) rabbit antibody (019-19741; Wako). Briefly, brain slices (30 μm thick) were incubated in 10% normal donkey or goat serum in PBST [1 x PBS (11189, Gibco) + 0.5% Tween 20 (P1379, Sigma)] containing 0.3% Triton X 100 (T8787, Sigma) for antigen retrieval and to block any background for 2 h at RT. Slices were incubated with the primary antibody (1:500) at 4°C overnight. After 3 washes with PBST, slices were incubated with the secondary antibody conjugated with Alexa Fluor 488 or Fluor 647 (1:500, Invitrogen) for 2 h at RT. After 5 washes with PBST, the sections were mounted, dehydrated, and cover-slipped. Images were acquired using Zeiss 880 inverted confocal Airy scan.

Zhang Z., Ma Q., Velagapudi R., Barclay W.E., Rodriguiz R.M., Wetsel W.C., Yang T., Shinohara M.L, & Terrando N. (2022). Annexin-A1 Tripeptide Attenuates Surgery-Induced Neuroinflammation and Memory Deficits Through Regulation the NLRP3 Inflammasome. Frontiers in Immunology, 13, 856254.

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Microglial Morphology Analysis via Immunostaining

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Immunostaining and analyses of microglial morphology were performed on the PFA-perfused slices using ionizing calcium-binding adaptor molecule 1 (Iba1) rabbit antibody (019-19741; Wako) . Briefly, brain slices (30 μm thick) were incubated in 10% normal donkey or goat serum in PBST [1 x PBS (11189, Gibco) + 0.5% Tween 20 (P1379, Sigma)] containing 0.3% Triton X 100 (T8787, Sigma) for antigen retrieval and to block any background for 2 hours at RT. Slices were incubated with the primary antibody (1:500) at 4°C overnight. After 3 washes with PBST, slices were incubated with the secondary antibody conjugated with Alexa Fluor 488 or Fluor 647
(1:500, Invitrogen) for 2 hours at RT. After 5 washes with PBST, the sections were mounted, dehydrated, and cover-slipped. Images were acquired using Zeiss 880 inverted confocal Airy scan.

Zhang Z., Ma Q., Velagapudi R., Barclay W.E., Rodriguiz R.M., Wetsel W.C., Shinohara M.L., & Terrando N. (2020). Annexin-A1 tripeptide attenuates surgery-induced neuroinflammation and memory deficits through regulation of the NLRP3 inflammasome.

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