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3 protocols using triclopyr

1

Triclopyr Preparation and Characterization

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Triclopyr (CAS: 55335-06-3, purity 99.9%; Product No 32016) was purchased from Sigma-Aldrich. The stock Triclopyr solutions were prepared with reconstituted water (2 mM CaCl2, 0.5 mM MgSO4, 0.75 mM NaHCO3, and 0.07 mM KCl in ultrapure water) according to the ISO 7346-1 guideline [22 ]. 3,4-Dichloroaniline (3,4-DCA) (CAS: 95-76-1, purity 98%; Product No 437778), Bradford’s reagent (Product No B6916), 5,5-dithio-bis-(2-nitrobenzoic acid) (CAS: 69-78-3; Product No D8130), acetylthiocholine iodide (CAS: 2260-50-6, purity ≥97%; Product No A7000), and dichlorvos (CAS: 62-73-7, purity 98.9%; Product No 45441) were purchased from Sigma-Aldrich.
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2

Immunofluorescence Staining of CRB3 Protein

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2,4-Dichlorophenoxyacetic acid (2,4-D) (CAS no. 94-75-7), para-aminohippuric acid (PAH) (CAS no. 61-78-9), triclopyr (CAS no. 55335-06-3), trimethylsilyldiazomethane (TMSD) (CAS no. 18107-18-1), N-tert-Butyldimethylsilyl-N-methyltrifluoroacetamide (MTBSTFA) (CAS no. 77377-52-7), ethyl acetate, toluene, lucifer yellow, pilocarpine, and other general laboratory chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-CRB3 rat primary monoclonal antibody was purchased from MyBioSource (Cat # MBS588104) and goat anti-rat IgG secondary antibody-Alexa Fluor 488 conjugate was purchased from Invitrogen (Cat # A11006).
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3

Determination of Iron Complexes

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Triclopyr (98% purity), EDDS trisodium salt solution (~ 35% in H2O) and analytical grade Fe(ClO4)3 were purchased from Sigma-Aldrich. Na2SO4 (99.9% purity) was purchased from Merck. Analytical grade H2SO4, NaCl and NaOH were supplied by Panreac. An acidic solution of Ti(IV) oxysulfate (Panreac) was used for H2O2 determination. L-ascorbic acid (99% purity)
and 1,10-phenanthroline monohydrate (99% purity) employed for the analysis of soluble iron were purchased from Sigma-Aldrich and Alfa-Aesar, respectively. Active chlorine was determined by reaction with N,N-diethyl-p-phenylenediamine (1:1) oxalate (99% purity)
provided by Acros Organics. Organic solvents of HPLC or analytical grade and other chemicals used, including analytical standards of carboxylic acids, were supplied by Panreac. Ultrapure
Milli-Q water (> 18.2 MΩ cm) was employed for the preparation of synthetic and analytical solutions. The combination of Fe(ClO4)3 and EDDS, both at a concentration of 0.03, 0.06 or 0.09 mM, gave rise to the corresponding Fe(III)-EDDS (1:1) complexes in solution, following the methodology described in earlier work (Ye et al. 2019a ).
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