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2 4 pyridinedicarboxylic acid 2 4 pdca

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2,4-pyridinedicarboxylic acid (2,4-PDCA) is a dicarboxylic acid compound with the chemical formula C7H5NO4. It is a white crystalline solid that is soluble in water and organic solvents. 2,4-PDCA is used as a chemical intermediate in various applications.

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Lab products found in correlation

Enzalutamide, by Selleck Chemicals (1 mentions) Calf thymus histone, by Merck Group (1 mentions) Complete edta free protease inhibitor cocktail, by Roche (1 mentions) Ripa buffer, by Merck Group (1 mentions) Doxorubicin, by Selleck Chemicals (1 mentions) Dc protein assay, by Bio-Rad (1 mentions) Isoproterenol hydrochloride, by Merck Group (1 mentions) Benserazide hydrochloride, by Merck Group (1 mentions) α ketoglutaric acid sodium salt, by Merck Group (1 mentions) Succinate glo jmjc demethylase hydroxylase assay kit, by Promega (1 mentions) Isoproterenol bitartrate, by Merck Group (1 mentions)

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2,4-PDCA (2 citations)

4 protocols using 2 4 pyridinedicarboxylic acid 2 4 pdca

1

Enzalutamide Assay with PDCA

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Enzalutamide was from Selleckchem (S1250). Dimethyl sulfoxide (DMSO) was from Fisher Scientific (BP231–1). 2,4-Pyridinedicarboxylic acid (2,4-PDCA) was purchased from Sigma-Aldrich (04473).
Paschalis A., Welti J., Neeb A.J., Yuan W., Figueiredo I., Pereira R., Ferreira A., Riisnaes R., Nava Rodrigues D., Jiménez-Vacas J.M., Kim S., Uo T., Di Micco P., Tumber A., Islam S., Moesser M.A., Abboud M., Kawamura A., Gurel B., Christova R., Gil V.S., Buroni L., Crespo M., Miranda S., Lambros M.B., Carreira S., Tunariu N., Alimonti A., Al-Lazikani B., Schofield C.J., Plymate S.R., Sharp A, & de Bono J.S. (2021). JMJD6 is a druggable oxygenase that regulates AR-V7 expression in prostate cancer. Cancer research, 81(4), 1087-1100.
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2

JARID Histone Demethylation Assay

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Purified human recombinant JARID1B/Plu-1 (BPS Bioscience) (240 ng), or JARID1D (Abnova) (240 ng) or KDM6B (Sigma-Aldrich) (80 ng) were added to 15 µl of a reaction mixture containing 5 µg of purified calf thymus histones (Sigma Aldrich) in 50 mM Tris HCl pH 7.5); 1 mM α-KG; 0.1 mM (NH4)2Fe(SO4)2; 2 mM ascorbate; protease inhibitors (Complete EDTA-free Protease Inhibitor Cocktail, Roche). The candidate inhibitor was added at indicated concentrations; 2,4-pyridinedicarboxylic acid (2,4-PDCA) (Sigma Aldrich) was at 5 µM. Reactions were incubated for 3 h at 37°C, stopped by 2× Laemmli loading buffer addition and directly loaded on gels for western blot analysis. Histone levels were quantified by coomassie blue stained H1 histones. Powder samples and DMSO stock solutions of 3195 were stored at −25°C
Mannironi C., Proietto M., Bufalieri F., Cundari E., Alagia A., Danovska S., Rinaldi T., Famiglini V., Coluccia A., La Regina G., Silvestri R, & Negri R. (2014). An High-Throughput In Vivo Screening System to Select H3K4-Specific Histone Demethylase Inhibitors. PLoS ONE, 9(1), e86002.
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3

Quantifying Caspase Cleavage and JARID1B Levels

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For measurement of caspase cleavage, UCDK9M4 cells were plated on tissue culture dishes and grown for 24 h before treatment with (DMSO) vehicle, doxorubicin (Selleck Chemicals, Houston, TX), or 2,4-pyridinedicarboxylic acid (2,4-PDCA) (Sigma–Aldrich, St. Louis, MO). Cells were incubated for an additional 24 hours following drug addition, then lysed as described below.
For measurement of JARID1B and cleaved-caspase 3, the indicated cells were lysed in RIPA buffer (Sigma–Aldrich) containing phosphatase and protease inhibitor cocktails, and 1 mM DTT for 20 min on ice. Lysates were then centrifuged for 15 min at 16000xg and the supernatant was collected. Protein concentration was measured using a DC protein assay (Bio Rad, Hercules, CA). A total of 40 μg of lysate (per well) was electrophoresed and transferred using standard protocols. Primary antibodies including anti-JARID1B, antitubulin, anti-actin, or anti-cleaved caspase 3 were used at a concentration of 1:500 to 1:2500. See antibody validation statements in Materials and MethodsSection 2.7 for catalogue numbers and supplier information.
Tobin S.J., Chang H., Kent M.S, & Davies A.E. (2021). JARID1-targeted histone H3 demethylase inhibitors exhibit anti-proliferative activity and overcome cisplatin resistance in canine oral melanoma cell lines. Veterinary and comparative oncology, 19(3), 518-528.
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4

Luminescent Enzymatic Assay for JmjC Demethylase Activity

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Ascorbate ((+)-sodium l-ascorbate), Fe(II) (ammonium iron(II) sulfate hexahydrate), and α-KG (α-ketoglutaric acid sodium salt) were from Sigma-Aldrich (St. Louis, MO). The inhibitor 2,4-pyridinedicarboxylic acid (2,4-PDCA) and the compounds (+)-isoproterenol (+)-bitartrate, benserazide hydrochloride, cafeic acid phenethyl ester, 7,8-dihydroxycoumarin, (-)-isoproterenol hydrochloride, and (+/-) taxifolin hydrate were also purchased from Sigma-Aldrich. The inhibitors 8-hydroxy-5-quinolinecarboxylic acid (IOX-1) and 5-chloro-2-[(E)-2[phenyl(pyridine-2-yl)methylidene] hydrazine-1-yl]pyridine (JIB 04) were purchased from TOCRIS (Bristol, UK). Daminozide and JHDM inhibitor III were purchased from EMD Millipore (Darmstadt, Germany). COSTAR 96-well half-area and 384-well low-volume white plates were purchased from Corning (Corning, NY).
The Succinate-Glo JmjC Demethylase/Hydroxylase Assay kit from Promega Corporation (Madison, WI) is composed of a succinate standard solution and components for two succinate detection reagents. Reagent I uses succinate as a substrate in a coupled enzyme reaction to produce ATP, and Reagent II detects ATP as light output from an ATP-dependent luciferase enzyme (Fig. 1).
Alves J., Vidugiris G., Goueli S.A, & Zegzouti H. (2018). Bioluminescent High-Throughput Succinate Detection Method for Monitoring the Activity of JMJC Histone Demethylases and Fe(II)/2-Oxoglutarate-Dependent Dioxygenases. SLAS discovery : advancing life sciences R & D, 23(3).
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