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Nmnat1

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NMNAT1 is an enzyme that catalyzes the conversion of nicotinamide mononucleotide (NMN) to nicotinamide adenine dinucleotide (NAD+) in the NAD+ biosynthesis pathway. It plays a crucial role in maintaining cellular NAD+ levels.

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Lab products found in correlation

Diaphorase, by Merck Group (4 mentions) Dmem f12, by Thermo Fisher Scientific (2 mentions) Quikchange site directed mutagenesis kit, by Agilent Technologies (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) α mem, by Thermo Fisher Scientific (2 mentions) Collagenase type 2, by Worthington (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (2 mentions) Glutamine, by Thermo Fisher Scientific (2 mentions) Histopaque 1077, by Merck Group (2 mentions) Ap20187, by Takara Bio (2 mentions) Celltracker cm dii dye, by Thermo Fisher Scientific (2 mentions) Recombinant mouse fcγr 1 and fcγr 4, by ACROBiosystems (2 mentions) Mabselect sure resin, by Merck Group (2 mentions) Superdex 200i 10x300gl column, by Merck Group (2 mentions) Recombinant human and mouse cd38 proteins, by R&D Systems (2 mentions) Anti mouse tnfα and il6 neutralizing antibodies, by R&D Systems (2 mentions) Luminex premixed multi analyte magnetic luminex assay kit, by R&D Systems (2 mentions) Olaparib, by LC Laboratories (2 mentions)

4 protocols using nmnat1

1

Detailed Multidisciplinary Research Protocol

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The following reagents were used: AP20187 (Clontech), 78c (Calbiochem), Olaparib (LC Laboratories), type II collagenase (Worthington Biochemical), QuikChange Site-Directed Mutagenesis Kit (Agilent Technologies), CellTracker CM-DiI dye (Invitrogen), recombinant mouse FcγR I and FcγR IV (Acro Biosystems), Isatuximab (TeneoBio), UniAb clone ID 337468 and 337469 (TeneoBio), MabSelect SuRe resin, Superdex 200i 10X300GL column, ADH, Diaphorase, 3TC, and HISTOPAQUE-1077 were from Sigma-Aldrich. Recombinant human and mouse CD38 proteins, NMNAT1, anti-mouse TNFα- and IL6-neutralizing antibodies, and Luminex Premixed Multi-Analyte Magnetic Luminex Assay Kit were from R&D Systems. αMEM, DMEM, DMEM:F12, fetal calf serum, glutamine, ITS, penicillin/streptomycin, and Lipofectamine 3000 were from Life Technologies. When not specified, reagents and chemicals were purchased from Sigma-Aldrich.
Chini C.C., Peclat T.R., Warner G.M., Kashyap S., Espindola-Netto J.M., de Oliveira G.C., Gomez L.S., Hogan K.A., Tarragó M.G., Puranik A.S., Agorrody G., Thompson K.L., Dang K., Clarke S., Childs B.G., Kanamori K.S., Witte M.A., Vidal P., Kirkland A.L., De Cecco M., Chellappa K., McReynolds M.R., Jankowski C., Tchkonia T., Kirkland J.L., Sedivy J.M., van Deursen J.M., Baker D.J., van Schooten W., Rabinowitz J.D., Baur J.A, & Chini E.N. (2020). CD38 ecto-enzyme in immune cells is induced during aging regulating NAD+ and NMN levels. Nature metabolism, 2(11), 1284-1304.
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2

Detailed Multidisciplinary Research Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
The following reagents were used: AP20187 (Clontech), 78c (Calbiochem), Olaparib (LC Laboratories), type II collagenase (Worthington Biochemical), QuikChange Site-Directed Mutagenesis Kit (Agilent Technologies), CellTracker CM-DiI dye (Invitrogen), recombinant mouse FcγR I and FcγR IV (Acro Biosystems), Isatuximab (TeneoBio), UniAb clone ID 337468 and 337469 (TeneoBio), MabSelect SuRe resin, Superdex 200i 10X300GL column, ADH, Diaphorase, 3TC, and HISTOPAQUE-1077 were from Sigma-Aldrich. Recombinant human and mouse CD38 proteins, NMNAT1, anti-mouse TNFα- and IL6-neutralizing antibodies, and Luminex Premixed Multi-Analyte Magnetic Luminex Assay Kit were from R&D Systems. αMEM, DMEM, DMEM:F12, fetal calf serum, glutamine, ITS, penicillin/streptomycin, and Lipofectamine 3000 were from Life Technologies. When not specified, reagents and chemicals were purchased from Sigma-Aldrich.
Chini C.C., Peclat T.R., Warner G.M., Kashyap S., Espindola-Netto J.M., de Oliveira G.C., Gomez L.S., Hogan K.A., Tarragó M.G., Puranik A.S., Agorrody G., Thompson K.L., Dang K., Clarke S., Childs B.G., Kanamori K.S., Witte M.A., Vidal P., Kirkland A.L., De Cecco M., Chellappa K., McReynolds M.R., Jankowski C., Tchkonia T., Kirkland J.L., Sedivy J.M., van Deursen J.M., Baker D.J., van Schooten W., Rabinowitz J.D., Baur J.A, & Chini E.N. (2020). CD38 ecto-enzyme in immune cells is induced during aging regulating NAD+ and NMN levels. Nature metabolism, 2(11), 1284-1304.
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3

Serum NMN Quantification by Fluorescent Assay

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To determine serum NMN levels, serum was deproteinated by addition of 10% trichloroacetic acid (TCA). Samples were centrifuged at 12,000 rpm for 2 minutes at 4°C. The supernatants were collected, and the pellets were re-suspended in 0.2 N NaOH for protein determination. TCA was removed with organic solvent (3 volumes 1,1,2-trichloro-1,2,2-trifluroethane: 1 volume trioctylamine) in a ratio of 2 volumes of organic solvent to 1 volume of sample. After phase separation, the top aqueous layer containing NMN was recovered and the pH was corrected by addition of 1M Tris pH 8.0. NMN concentration was measured by fluorescence of resazurin redox reaction using a coupled assay with NMNAT1 (R&D −5865NT), ADH and Diaphorase (Sigma-Aldrich).
Chini C.C., Peclat T.R., Warner G.M., Kashyap S., Espindola-Netto J.M., de Oliveira G.C., Gomez L.S., Hogan K.A., Tarragó M.G., Puranik A.S., Agorrody G., Thompson K.L., Dang K., Clarke S., Childs B.G., Kanamori K.S., Witte M.A., Vidal P., Kirkland A.L., De Cecco M., Chellappa K., McReynolds M.R., Jankowski C., Tchkonia T., Kirkland J.L., Sedivy J.M., van Deursen J.M., Baker D.J., van Schooten W., Rabinowitz J.D., Baur J.A, & Chini E.N. (2020). CD38 ecto-enzyme in immune cells is induced during aging regulating NAD+ and NMN levels. Nature metabolism, 2(11), 1284-1304.
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4

Serum NMN Quantification by Fluorescent Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
To determine serum NMN levels, serum was deproteinated by addition of 10% trichloroacetic acid (TCA). Samples were centrifuged at 12,000 rpm for 2 minutes at 4°C. The supernatants were collected, and the pellets were re-suspended in 0.2 N NaOH for protein determination. TCA was removed with organic solvent (3 volumes 1,1,2-trichloro-1,2,2-trifluroethane: 1 volume trioctylamine) in a ratio of 2 volumes of organic solvent to 1 volume of sample. After phase separation, the top aqueous layer containing NMN was recovered and the pH was corrected by addition of 1M Tris pH 8.0. NMN concentration was measured by fluorescence of resazurin redox reaction using a coupled assay with NMNAT1 (R&D −5865NT), ADH and Diaphorase (Sigma-Aldrich).
Chini C.C., Peclat T.R., Warner G.M., Kashyap S., Espindola-Netto J.M., de Oliveira G.C., Gomez L.S., Hogan K.A., Tarragó M.G., Puranik A.S., Agorrody G., Thompson K.L., Dang K., Clarke S., Childs B.G., Kanamori K.S., Witte M.A., Vidal P., Kirkland A.L., De Cecco M., Chellappa K., McReynolds M.R., Jankowski C., Tchkonia T., Kirkland J.L., Sedivy J.M., van Deursen J.M., Baker D.J., van Schooten W., Rabinowitz J.D., Baur J.A, & Chini E.N. (2020). CD38 ecto-enzyme in immune cells is induced during aging regulating NAD+ and NMN levels. Nature metabolism, 2(11), 1284-1304.
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