Pheophorbide a

Manufactured by Merck Group

Sourced in United States, Germany

Pheophorbide A is a chlorophyll derivative that is used as a laboratory reagent. It is a dark green crystalline solid with a molecular formula of C35H36N4O5. Pheophorbide A is primarily used as a standard compound for the analysis and identification of chlorophyll and chlorophyll-related compounds.

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Pheophorbide a (PhA) (2 citations)

5 protocols using pheophorbide a

Porphyrin Biosynthesis Assay

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Sodium orthovanadate, N-ethylmaleimide (NEM), EDTA, EGTA, pheophorbide-A, haemin, prazosin, coproporphyrinogen-III and protoporphyrin-IX were purchased from Sigma–Aldrich. 8-Azido-[α-³²P]ATP was obtained from Izinta.

Kiss K., Kucsma N., Brozik A., Tusnady G.E., Bergam P., vanNiel G, & Szakacs G. (2015). Role of the N-terminal transmembrane domain in the endo-lysosomal targeting and function of the human ABCB6 protein. Biochemical Journal, 467(Pt 1), 127-139.

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ABCG2 Transporter Inhibition Assay

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All chemicals were of molecular biology grade from Sigma‐Aldrich (St. Louis, MO, USA), unless stated otherwise. Mitoxantrone, rhodamine 123, pheophorbide A, Ko143, potassium antimony (III) tartrate hydrate, adenosine 5′‐triphosphate disodium salt (ATP), ammonium molybdate, ascorbic acid, and ouabain octahydrate were from Sigma‐Aldrich. Poly‐ethyleneimine was from Polysciences Europe (Eppelheim, Germany). Sodium orthovanadate was from New England Biolabs (Frankfurt, Germany). G418 was from Santa Cruz Biotechnology (Dallax, TX, USA). Oligonucleotides for site‐directed mutagenesis were purchased from Eurofins (Munich, Germany). Monoclonal mouse anti‐ABCG2 (BXP‐21) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit anti‐β‐actin (D6A8) was purchased from Cell Signaling (Danvers, MA, USA). Phusion DNA polymerase and DpnI are from NEB (New England Biolabs, Ipswich, MA, USA).

Khunweeraphong N, & Kuchler K. (2020). The first intracellular loop is essential for the catalytic cycle of the human ABCG2 multidrug resistance transporter. Febs Letters, 594(23), 4059-4075.

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Fluorescence-Based Functional Assays for ABC Transporters

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Calcein acetoxymethyl ester (calcein AM; Molecular Probes, Eugene, OR, USA) accumulation and pheophorbide A (Sigma-Aldrich, St. Louis, MO, USA) accumulation were used as functional assays of ABCB1 and ABCG2, respectively [31 (link)]. Cells were loaded with appropriate probe and then analyzed by flow cytometry (Cytomics FC500; Beckman Coulter, Indianapolis, IN, USA), as described previously [31 (link)].

Pastvova N., Dolezel P, & Mlejnek P. (2021). Heat Shock Protein Inhibitor 17-Allyamino-17-Demethoxygeldanamycin, a Potent Inductor of Apoptosis in Human Glioma Tumor Cell Lines, Is a Weak Substrate for ABCB1 and ABCG2 Transporters. Pharmaceuticals, 14(2), 107.

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HPLC-HRMS Analysis of Plant Compounds

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For HPLC-HRMS analyses, H₂O was purified with a Barnstead EASYpure RF compact ultrapure water system, and CH₃OH (VWR International, Rosny-sous-Bois-cedex, France) and HCOOH (Sigma Aldrich, Steinheim, Germany) were LC-MS grade. The other solvents were of analytical grade. Amentoflavone, luteolin and luteolin-7-glucoside were purchased from Carl Roth (Karlsruhe, Germany). Apigenin and pheophorbide A were purchased from Sigma Aldrich (Steinheim, Germany), and podocarpusflavone A, from Carbosynth (Berkshire, UK). Silica gel 60 for VLC (particle size 0.040–0.063 mm) and LiChroprep^® RP-18 (0.040–0.063 mm) for RP-18 column chromatography were purchased from Merck (Darmstadt, Germany). For solid-phase extraction, 5 g or 10 g Isolute^® C18 (EC) cartridges (Biotage, Uppsala, Sweden) were used.

Pferschy-Wenzig E.M., Ortmann S., Atanasov A.G., Hellauer K., Hartler J., Kunert O., Gold-Binder M., Ladurner A., Heiß E.H., Latkolik S., Zhao Y.M., Raab P., Monschein M., Trummer N., Samuel B., Crockett S., Miao J.H., Thallinger G.G., Bochkov V., Dirsch V.M, & Bauer R. (2022). Characterization of Constituents with Potential Anti-Inflammatory Activity in Chinese Lonicera Species by UHPLC-HRMS Based Metabolite Profiling. Metabolites, 12(4), 288.

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BCRP Transporter Inhibition Assay

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Bodipy prazosin (Invitrogen Life technologies) and pheophorbide A (Sigma Aldrich) were used as fluorescent dyes and substrates of BCRP. Inhibitors of ABC transporters described in Table 1 were purchased from Sigma Aldrich and Tebu-bio for Ko134.

Ben Cheikh Y., Xuereb B., Boulangé-Lecomte C, & Le Foll F. (2018). Multixenobiotic resistance in Mytilus edulis: Molecular and functional characterization of an ABCG2- type transporter in hemocytes and gills. Aquatic toxicology (Amsterdam, Netherlands), 195.

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