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R9504

Manufactured by Merck Group
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Sourced in Germany

R9504 is a laboratory equipment product manufactured by Merck Group. It is designed for use in various scientific and research applications that require precise and reliable measurements. The core function of R9504 is to provide accurate and consistent data collection capabilities. Detailed technical specifications and intended use cases are not available.

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Lab products found in correlation

A0737, by Merck Group (2 mentions) Lab tek 2 chamber slide system, by Thermo Fisher Scientific (1 mentions) Niacinamide, by Merck Group (1 mentions) Pyridoxine hydrochloride, by Merck Group (1 mentions) Thiamine, by Merck Group (1 mentions) Y1251, by Merck Group (1 mentions) Lp0021, by Thermo Fisher Scientific (1 mentions) C11995500bt, by Thermo Fisher Scientific (1 mentions)

4 protocols using r9504

1

iPSCs and iPSC-derived MNs treatment

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Ctrl and RTD iPSCs and iPSC-derived MNs were treated with 10 µM RF (R9504, Merck KGaA, Germany) overnight or 100 µM NAC amide (A0737, Merck KGaA, Germany) overnight. For combined treatment, 10 µM RF and 100 µM NAC were administered together.
Niceforo A., Marioli C., Colasuonno F., Petrini S., Massey K., Tartaglia M., Bertini E., Moreno S, & Compagnucci C. (2021). Altered cytoskeletal arrangement in induced pluripotent stem cells and motor neurons from patients with riboflavin transporter deficiency. Disease Models & Mechanisms, 14(2), dmm046391.
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2

Ultrastructural Analysis of Treated Cells

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Ctrl and RTD MNs were plated in a chamber slide (Lab-Tek II Chamber Slide System, ThermoFisher Scientific). Some samples were treated with 1 µM RF (R9504, Merck KGaA, Darmstadt, Germany) and 100 µM N-acetylcysteine amide (NAC) (A0737, Merck KGaA) overnight.
Treated and untreated cells were fixed and embedded in epoxy resin, according to Colasuonno and coll. [11 (link)]. Electron microscopic observations and analyses were performed by a DualBeam focused ion beam/scanning electron microscopy (FIB/SEM, Helios Nanolab, FEI, Hillsboro, OR, USA). Random images for statistical analysis purposes were electronically captured. Representative images were composed in an Adobe Photoshop CS6 format.
Colasuonno F., Bertini E., Tartaglia M., Compagnucci C, & Moreno S. (2020). Mitochondrial Abnormalities in Induced Pluripotent Stem Cells-Derived Motor Neurons from Patients with Riboflavin Transporter Deficiency. Antioxidants, 9(12), 1252.
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3

Isotopic Labeling of R15B Protein

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Isotopically labelled R15B414-613 was overexpressed in M9 media (6 g/l Na2HPO4, 3 g/l KH2PO4, 0.5 g/l NaCl) supplemented with 1.7 g/l yeast nitrogen base without NH4Cl and amino acids (Sigma-Aldrich, Y1251). 1 g/l 15NH4Cl and 4 g/l 13C-glucose were supplemented for 15N and 13C labelling respectively. Additionally, filtered-sterile solutions of 2 mM MgSO4 (final concentration in M9 media), 25 mg FeSO4.7H2O, and 1 ml vitamin mix (100 ml vitamin mix stock solution contains riboflavin 100 mg (Sigma-Aldrich, R9504), niacinamide 100 mg (Sigma-Aldrich, N5535), pyridoxine hydrochloride 10 mg (Sigma-Aldrich, P9755), thiamine 100 mg (Sigma-Aldrich, T1270) were added to the media. Cells were grown in M9 media as described for LB media above with the following exception: cells were grown at 37 °C until OD600 0.6 before induction of protein expression with 200 μM IPTG for 18 hours at 22 °C. Isotopically labelled protein was purified as described above for the native protein.
Fatalska A., Hodgson G., Freund S.M., Maslen S.L., Morgan T., Thorkelsson S.R., van Slegtenhorst M., Lorenz S., Andreeva A., Kaat L.D, & Bertolotti A. (2023). Recruitment of trimeric eIF2 by phosphatase non-catalytic subunit PPP1R15B. Molecular cell, 84(3), 506-521.e11.
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4

Bacterial Strains and Plasmid Engineering for EHEC O157:H7 Studies

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The bacterial strains and plasmids used in this study are listed in SI Appendix, Table S1. Mutant strains of EHEC O157:H7 EDL933 were generated using the λ-Red recombination system. Complementary strains were established by cloning rbfS and rbfR and native promoters into the pACYC-184 plasmid and transformed into EHEC O157:H7 EDL933 and C. rodentium DBS100. The strain for RbfR and RbfS purification was generated by cloning rbfR and rbfS into the pET28a plasmid. Bacterial strains were grown in Luria-Bertani (LB) broth (Oxoid; LP0021 and LP0042), Dulbecco’s modified Eagle medium (DMEM, Gibco; C11995500BT), or simulated colonic environment medium (SCEM, 6.25 g/L tryptone, 2.6 g/L D-glucose, 0.88 g/L NaCl, 2.7 g/L KHCO3, 0.43 g/L KH2PO4, 1.7 g/L NaHCO3 and 4.0 g/L bile salts) (49 ) with different concentrations of riboflavin (Sigma; R9504). Bacteria were inoculated into a fresh medium and grown at 37°C under shaking. When needed, isopropyl β-d-thiogalactoside (IPTG) and antibiotics were added to the medium at the following final concentrations: 0.1 mM IPTG, 100 μg/ml ampicillin, 25 μg/ml chloramphenicol, 50 μg/ml kanamycin, and 10 μg/ml tetracycline. The primers used for all manipulations are listed in SI Appendix, Table S1.
Liu B., Liu Y., Yang B., Wang Q., Liu X., Qin J., Zhao K., Li F., Feng X., Li L., Wu P., Liu M., Zhu S., Feng L, & Wang L. (2022). Escherichia coli O157:H7 senses microbiota-produced riboflavin to increase its virulence in the gut. Proceedings of the National Academy of Sciences of the United States of America, 119(48), e2212436119.
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