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4 protocols using acetate tetrahydrate

1

Synthesis of Transition Metal Complexes

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Nickel(II) acetate tetrahydrate
(98%), cobalt(II) acetate tetrahydrate
(99%), copper(II) dichloride dihydrate (99%), manganese(II) acetate
tetrahydrate (99%), iron(II) chloride tetrahydrate (99%), and 2-mercaptopyridine N-oxide sodium salt (96%) were all purchased from Sigma-Aldrich
and used as received.
Acetonitrile (CH3CN; Fulltime
Chemical, HPLC/Spectro
grade, >99%) was dried with 4 Å molecular sieves (Alfa Aesar)
before usage. Tetrabutylammonium hexafluorophosphate (NBu4PF6; Tokyo Chemical Industry Co., Ltd.,
>98.0%) and glacial acetic acid (CH3COOH; Merck, 100%)
were used as received.
Electrospray ionization (ESI) mass spectrometry
was done using
a Finnigan MAT LCQ spectrometer while high-resolution mass spectroscopy
(HRMS) was done using a Bruker micrOTOF-QII spectrometer. Elemental
analyses (EAL) of carbon, hydrogen, nitrogen, and nickel were done
using an ElemenVario Micro Cube. NMR spectra were performed on Bruker
AV III 400HD (BBFO probe) spectrometer. 1H NMR spectra
chemical shifts were reported in δ ppm relative to DMSO-d6 (δ = 2.05 ppm). Multiplicities were
recorded as: d (doublet) and t (triplet) respectively. The number
of protons (n) for a given resonance was indicated
by nH while coupling constants were reported as J value in Hertz (Hz).
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2

Synthesis and Characterization of CoMoO4 Nanocomposites

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Molybdenum(ii) acetate (Mo2(O2CCH3)4), cobalt(ii) acetate tetrahydrate (C4H6CoO4·4H2O), ethanol (C2H5OH), methanol (CH3OH), ethylene glycol (C2H6O2) and citric acid (C6H8O7) were purchased from Merck Chemicals, Ltd. Double-distilled and deionized water was used throughout the experiment. The phytochemical extract of E. cognata plant leaves was used as reducing and stabilizing agents in the synthesis of CoMoO4 and was sampled from Rawalakot AJK Pakistan. Acetylene black, polyvinylidenedifluoride (PVDF) and N-methyl pyrrolidinone (NMP) were used in the fabrication of the electrode.
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3

Synthesis of Metal Complexes

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Potassium ethyl xanthogente (96%, Sigma-Aldrich), cupric chloride dihydrate (97%, Saarchem), cobalt(ii) acetate tetrahydrate (98%, Saarchem); and nickel(ii) acetate tetrahydrate, chloroform (min 99%), acetone and hexane, purchased from Merck Chemicals. All chemicals were used as received with no further purification.
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4

Detecting Protein Interactions and Modifications

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The following antibodies were purchased from Abcam (Cambridge, CA): rabbit polyclonal antibodies against GFP, HA, and PD‐L1 (Alexa Fluor 488), a monoclonal antibody against FLAG. Rabbit polyclonal anti‐pSTAT (Tyr701) was obtained from BD Bioscience (San Jose, CA). For immunofluorescence staining, Alexa Fluor‐488, ‐546, and ‐647 goat anti‐rabbit or anti‐mouse secondary antibodies (Thermo Fisher Scientific, Waltham, MA) were used. For western blot detection, IRDye 700CW and IRDye 800CW anti‐rabbit or anti‐mouse secondary antibodies (LI‐COR Bioscience, Lincoln, NE) were used. Recombinant human IFN‐γ and myoricin were purchased from Merck (Darmstadt, Germany). Sulfo‐Cy3‐DBCO was obtained from click chemistry tools (Scottsdale, AZ). ATP disodium hydrate salt and acetate tetrahydrate were purchased from Sigma Aldrich (Saint Louis, MO). Zaragozic acid A trisodium salt was purchased from Sta. Cruz Biotechnology (Dallas, TX). MβCD was obtained from Sigma Aldrich (San Luis, MO). For transfection FugeneHD from Promega (Madison, WI) and Opti‐MEM from Thermo Fisher Scientific (Waltham, MA) were used. For immunoprecipitation Protein A Sepharose Fast Flow Beads were purchased from GE Healthcare (Chicago, IL).
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