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Fam dye

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FAM Dye is a fluorescent dye used as a reporter molecule in various molecular biology applications, including real-time PCR and DNA sequencing. It has an excitation wavelength of approximately 494 nm and an emission wavelength of approximately 518 nm.

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Lab products found in correlation

Taqman fast universal pcr master mix, by Thermo Fisher Scientific (9 mentions) Superscript 3 rt pcr system, by Thermo Fisher Scientific (7 mentions) Rneasy kit, by Qiagen (4 mentions) Quick rna miniprep kit, by Zymo Research (4 mentions) Random hexamer primer, by Thermo Fisher Scientific (3 mentions) Gapd vic mgb dye, by Thermo Fisher Scientific (2 mentions) First strand cdna synthesis kit, by Qiagen (2 mentions) Abi prism 7700, by Thermo Fisher Scientific (2 mentions) Rnaeasy mini kit, by Qiagen (2 mentions) 2100 bioanalyzer, by Agilent Technologies (2 mentions) Superscript 2 rt pcr system, by Thermo Fisher Scientific (2 mentions) Actin vic mgb dye, by Thermo Fisher Scientific (2 mentions) Turbo dna free, by Thermo Fisher Scientific (1 mentions) Superscript 2 rt, by Thermo Fisher Scientific (1 mentions) Rneasy plus micro kit, by Qiagen (1 mentions) Lightcycler 1.5 instrument, by Roche (1 mentions) Custom taqman gene expression assay service, by Thermo Fisher Scientific (1 mentions) Lightcycler taqman master kit, by Roche (1 mentions) Rneasy mini kit, by Qiagen (1 mentions)

Close spelling variants of this product

FAM dye-labeled primers FAM dye labeled probes FAM reporter dye FAM dye-labeled TaqMan probes TaqMan MGB probe (6-FAM dye-labeled), FAM dye-labeled MGB-probes TaqMan Gene Expression Assays (FAM dye-labeled MGB probe; TaqMan gene expression assays FAM-dye labeled FAM™ dye label format FAM dye-labeled TaqMan assays

13 protocols using fam dye

1

Regulatory T Cell Gene Expression

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Total RNA was isolated from sorted cells with RNeasy kit (Qiagen). Reverse transcription was performed with the SuperScript III RT-PCR system (Invitrogen) and quantitative real-time reverse transcription (RT)-PCR with Taqman® Fast Universal PCR master mix, internal house keeping gene mouse (GAPD VIC-MGB dye) and specific target gene primers (FAM Dye) (Applied Biosystems) on Step-One- Plus machine. Relative expression was normalized to GAPD for Notch1-4 receptor and calculated as fold change compared to wild-type CD4+GFP− conventional T cells for Pofut1, Notch1, RBPJ and Rictor regarding the regulatory T cell-specific deficiency and fold change normalized to wild-type Treg cells for IFNg and IL12rb2.
Charbonnier L.M., Wang S., Georgiev P., Sefik E, & Chatila T.A. (2015). Control of peripheral tolerance by regulatory T cell-intrinsic Notch signaling. Nature immunology, 16(11), 1162-1173.
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2

Quantitative gene expression analysis

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The tissue was thawed and mRNA was isolated using the Qiagen RNAeasy mini kit (Qiagen) as per manufacturers’ instructions. RNA concentration was confirmed using a Bioanalyzer 2100 (Agilent Technologies, Palo Alto, CA) and cDNA was made from 1 µg of total RNA using the first strand cDNA synthesis kit (SABiosciences, Frederick, MD). One microgram of cDNA used for real-time PCR analysis (ABI PRISM 7700, Applied Biosystems Inc., Waltham, MA) utilizing the primer probe sets with the Fam dye (Applied Biosystems) (Foxp3, CCL22, Granzyme B, CCL5, IFN-γ, CXCL9, CXCL10, CXCL11 and IFN-γ). Data analysis was carried out with the instrument software and the ΔΔCt method was used with normalization of the raw data to an external RNA control.
Budiu R.A., Vlad A.M., Nazario L., Bathula C., Cooper K.L., Edmed J., Thaker P.H., Urban J., Kalinski P., Lee A.V., Elishaev E.L., Conrads T.P, & Flint M.S. (2016). Restraint and Social Isolation Stressors Differentially Regulate Adaptive Immunity and Tumor Angiogenesis in a Breast Cancer Mouse Model. Cancer and clinical oncology, 6(1), 12-24.
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3

Quantitative RT-PCR Analysis of Murine Treg Cells

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RNA was extracted from cells using Quick-RNA MiniPrep kit (Zymo Research) according to the manufacturer protocol. Reverse transcription was performed with the SuperScript III RT-PCR system and random hexamer primers (Invitrogen) and quantitative real-time reverse transcription (RT)-PCR with Taqman® Fast Universal PCR master mix, internal house keeping gene mouse (Hprt VIC-MGB dye) and specific target gene primers for murine Rorc, as indicated (FAM Dye) (Applied Biosystems) on Step-One-Plus machine. Relative expression was normalized to Hprt and calculated as fold change compared to Foxp3YFPCre Treg cells.
Abdel-Gadir A., Stephen-Victor E., Gerber G.K., Rivas M.N., Wang S., Harb H., Wang L., Li N., Crestani E., Spielman S., Secor W., Biehl H., Dibendetto N., Dong X., Umetsu D.T., Bry L., Rachid R, & Chatila T.A. (2019). Microbiota Therapy Acts Via a Regulatory T Cell MyD88/RORγt Pathway to Suppress Food Allergy. Nature medicine, 25(7), 1164-1174.
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4

RNA Isolation and RT-qPCR Analysis

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RNA was extracted from third-instar larval CNS or adult heads (5-7 days post-pupation) with RNeasy Plus Micro kit (Qiagen). RNA isolation was followed with DNase digestion with Turbo DNA-free (Ambion). For the first strand synthesis Super Script II RT was used (Invitrogen). Taqman Fast Universal PCR solution was mixed with TaqMan probe with an Applied Biosystems FAM dye. RPL32 was amplified as an internal control. Expression fold-changes are quantified by ddCT method. Data represent three biological and three technical replicates.
Genç Ö., An J.Y., Fetter R.D., Kulik Y., Zunino G., Sanders S.J, & Davis G.W. (2020). Homeostatic plasticity fails at the intersection of autism-gene mutations and a novel class of common genetic modifiers. eLife, 9, e55775.
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5

Gene Expression Analysis in Treated Cells

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RNA was extracted from treated cells, as mentioned in the figure legends, using Quick-RNA MiniPrep kit (Zymo Research) according to the manufacturer protocol. Reverse transcription was performed with the SuperScript III RTPCR system (Invitrogen) and quantitative real-time reverse transcription (RT)-PCR with Taqman® Fast Universal PCR master mix, internal house keeping gene mouse (GAPDH VIC-MGB dye) and specific target gene primers for murine Il6, Ccl11, Rorc or Il4 genes, as indicated (FAM Dye) (Applied Biosystems) on Step-One-Plus machine. Relative expression was normalized to GAPDH and calculated as fold change compared to un-stimulated WT cells.
Massoud A.H., Charbonnier L.M., Lopez D., Pellegrini M., Phipatanakul W, & Chatila T.A. (2016). An asthma-associated IL4R variant exacerbates airway inflammation by promoting conversion of regulatory T cells to TH17-like cells. Nature medicine, 22(9), 1013-1022.
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6

Glycolysis and Metabolism Gene Expression

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Total RNA was isolated from sorted cells with RNeasy kit (Qiagen). Reverse transcription was performed with the
SuperScript II RT-PCR system (Invitrogen) and quantitative real-time reverse transcription (RT)-PCR with Taqman®Fast Universal PCR master mix, internal house-keeping gene mouse (Actin VIC-MGB dye) and specific target gene
primers (FAM Dye) (Applied Biosystems) on Step-One-Plus machine. Relative expression was normalized to Actin for genes encoding
for the enzymes of the glycolysis cascade (Scl2a3, Hk1, Hk2, Gpi, Pfkfb3,
Aldoa, Gapdh, Tpi, Pdk1, Pgam1, Pgam5, Eno1, Pkm2,
Ldha, Scl16a1), the pentose phosphate cycle (G6pdx, Pgd,
Rpia, Rpe), Foxp3, Rictor, Rptor, HIF1a and
Myc and calculated as fold change compared to WT CD4+GFP– Teff cells or WT
CD4+GFP– Treg cells isolated from Foxp3EGFP mice.
Charbonnier L.M., Cui Y., Victor E., Harb H., Lopez D., Bleesing J.J., Garcia-Lloret M.I., Chen K., Ozen A., Carmeliet P., Li M.O., Pellegrini M, & Chatila T.A. (2019). Functional Reprogramming of Regulatory T cells in the absence of Foxp3. Nature immunology, 20(9), 1208-1219.
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7

Glycolysis and Metabolism Gene Expression

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Total RNA was isolated from sorted cells with RNeasy kit (Qiagen). Reverse transcription was performed with the
SuperScript II RT-PCR system (Invitrogen) and quantitative real-time reverse transcription (RT)-PCR with Taqman®Fast Universal PCR master mix, internal house-keeping gene mouse (Actin VIC-MGB dye) and specific target gene
primers (FAM Dye) (Applied Biosystems) on Step-One-Plus machine. Relative expression was normalized to Actin for genes encoding
for the enzymes of the glycolysis cascade (Scl2a3, Hk1, Hk2, Gpi, Pfkfb3,
Aldoa, Gapdh, Tpi, Pdk1, Pgam1, Pgam5, Eno1, Pkm2,
Ldha, Scl16a1), the pentose phosphate cycle (G6pdx, Pgd,
Rpia, Rpe), Foxp3, Rictor, Rptor, HIF1a and
Myc and calculated as fold change compared to WT CD4+GFP– Teff cells or WT
CD4+GFP– Treg cells isolated from Foxp3EGFP mice.
Charbonnier L.M., Cui Y., Victor E., Harb H., Lopez D., Bleesing J.J., Garcia-Lloret M.I., Chen K., Ozen A., Carmeliet P., Li M.O., Pellegrini M, & Chatila T.A. (2019). Functional Reprogramming of Regulatory T cells in the absence of Foxp3. Nature immunology, 20(9), 1208-1219.
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8

Regulatory T Cell Gene Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total RNA was isolated from sorted cells with RNeasy kit (Qiagen). Reverse transcription was performed with the SuperScript III RT-PCR system (Invitrogen) and quantitative real-time reverse transcription (RT)-PCR with Taqman® Fast Universal PCR master mix, internal house keeping gene mouse (GAPD VIC-MGB dye) and specific target gene primers (FAM Dye) (Applied Biosystems) on Step-One- Plus machine. Relative expression was normalized to GAPD for Notch1-4 receptor and calculated as fold change compared to wild-type CD4+GFP− conventional T cells for Pofut1, Notch1, RBPJ and Rictor regarding the regulatory T cell-specific deficiency and fold change normalized to wild-type Treg cells for IFNg and IL12rb2.
Charbonnier L.M., Wang S., Georgiev P., Sefik E, & Chatila T.A. (2015). Control of peripheral tolerance by regulatory T cell-intrinsic Notch signaling. Nature immunology, 16(11), 1162-1173.
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9

Quantitative gene expression analysis

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The tissue was thawed and mRNA was isolated using the Qiagen RNAeasy mini kit (Qiagen) as per manufacturers’ instructions. RNA concentration was confirmed using a Bioanalyzer 2100 (Agilent Technologies, Palo Alto, CA) and cDNA was made from 1 µg of total RNA using the first strand cDNA synthesis kit (SABiosciences, Frederick, MD). One microgram of cDNA used for real-time PCR analysis (ABI PRISM 7700, Applied Biosystems Inc., Waltham, MA) utilizing the primer probe sets with the Fam dye (Applied Biosystems) (Foxp3, CCL22, Granzyme B, CCL5, IFN-γ, CXCL9, CXCL10, CXCL11 and IFN-γ). Data analysis was carried out with the instrument software and the ΔΔCt method was used with normalization of the raw data to an external RNA control.
Budiu R.A., Vlad A.M., Nazario L., Bathula C., Cooper K.L., Edmed J., Thaker P.H., Urban J., Kalinski P., Lee A.V., Elishaev E.L., Conrads T.P, & Flint M.S. (2016). Restraint and Social Isolation Stressors Differentially Regulate Adaptive Immunity and Tumor Angiogenesis in a Breast Cancer Mouse Model. Cancer and clinical oncology, 6(1), 12-24.
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10

Quantitative Real-Time PCR Analysis of Reelin and NR2B

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Quantitative Real-Time PCR was performed using the LightCycler 1.5 Instrument from Roche (Roche Applied Science, Mannheim, Germany). For the reaction mix the LightCycler TaqMan Master kit (Roche) was used according to the manufacturers instructions. Primer and probe reagents were ready-made reagents using FAM-dye (Pre-designed TaqMan Assay Reagents; Applied Biosystems) with the following assay- and corresponding GenBank accession numbers: Hprt1 Rn01527840_m1/NM_012583.2, Reelin Rn00589609_m1/NM_080394.2. For NR2B analysis the reagent was designed from the Custom TaqMan Gene Expression Assay Service (Applied Biosystems) with following primer- and probe sequences: GluN2B-237s fw 5′-CAAGCCTGGCATGGTCTTCT-3′, rev 5′-GGATTGGCGCTCCTCTATGG-3′, probe 5′-FAM-CCATCAGCAGAGGTATCT-NFQ-3′ (M91562.1). qRT-PCR reactions were started with an initial denaturation step for 10 min at 95°C, followed by 45 cycles of 95°C for 10 sec, 60°C for 60 sec and 72°C for 1 sec. Relative amounts of Reelin and GluN2B had been normalized with Hprt for mRNA amount variations. The mRNA expression is presented as the change of relative quantities and was analyzed using the 2-ΔΔCt method [24] (link).
Landgraf P., Mikhaylova M., Macharadze T., Borutzki C., Zenclussen A.C., Wahle P, & Kreutz M.R. (2014). Binding of Y-P30 to Syndecan 2/3 Regulates the Nuclear Localization of CASK. PLoS ONE, 9(2), e85924.
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