Tyra23

Manufactured by Merck Group

Sourced in China

TyrA23 is a laboratory equipment product manufactured by Merck Group. It is designed for analytical and experimental applications. The core function of TyrA23 is to perform specific tasks related to the analysis and processing of samples. No further details about the intended use or capabilities of this product are available.

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Lab products found in correlation

6 protocols using tyra23

Arabidopsis Chemical Treatments Protocol

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BRZ, BFA, TyrA23, TyrA51, and PPMP were obtained from Sigma-Aldrich and used from DMSO stock solutions (20 mM for BRZ, 50 mM for BFA, TyrA23, TyrA51, and 10 mM for PPMP) and diluted with 1/23 MS growth medium. The hormone eBL (Sigma-Aldrich) was dissolved in 85% EtOH to yield a 1 mM stock solution. The sterol-disrupting reagent MbCD (Sigma-Aldrich) was dissolved in deionized water to yield a 200 mM stock solution. For long-term treatments, eBL or BRZ was dissolved in 1/23 MS solid medium onto which roots were placed. Short-term treatments were done by mixing the hormone or drug into liquid 1/23 MS growth medium and incubating Arabidopsis seedlings. The TyrA23, MbCD, and PPMP treatments were performed as described (Li et al., 2011) .

Wang L., Li H., Lv X., Chen T., Li R., Xue Y., Jiang J., Jin B., Baluška F., Šamaj J., Wang X, & Lin J. (2015). Spatiotemporal Dynamics of the BRI1 Receptor and its Regulation by Membrane Microdomains in Living Arabidopsis Cells. Molecular plant, 8(9).

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Molecular Tools for Plant Signaling Analysis

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LaCl₃ (purity >99.99%) was purchased from Aladdin Bio-Chem Technology Co. Ltd (Shanghai, China). FM4-64 was purchased from Thermo Fisher Scientific Co. Ltd (Shanghai, China). DPI, TyrA23, Murashige and Skoog medium (MS), methyl 1-(butylcarbamoyl)-2-benzimidazolecarbamate (Benomyl), 6-benzylaminopurine, indole acetic acid (IAA), and JA were purchased from Sigma-Aldrich Co. Ltd (Shanghai, China). Flg22 was purchased from MedChemExpress Co. Ltd (Shanghai, China). Anti-plant actin mouse monoclonal antibody (A01050) was purchased from Abbkine Scientific Co. Ltd (Wuhan, China). Goat anti-mouse IgG H&L [horseradish peroxidase (HRP)] (ab205719) and HRP anti-GFP antibody (ab6663) were purchased from Abcam Co. Ltd (Shanghai, China). Other chemicals used in this study were analytical reagents and were purchased from Sinopharm Chemical Reagent Co. Ltd. (Shanghai, China).

Cheng M., Wang L., Zhou Q., Chao D., Nagawa S., He D., Zhang J., Li H., Tan L., Gu Z., Huang X, & Yang Z. (2021). Lanthanum(III) triggers AtrbohD- and jasmonic acid-dependent systemic endocytosis in plants. Nature Communications, 12, 4327.

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Imaging Dyes and Inhibitors Stock Solutions

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Styryl dye FM 4–64 (Molecular Probes, catalogue number T13320), BFA, TYR A23, TYR A51, WM, FIL, DNS, cytochalasin D, latrunculin B, ORY, TX and auxins 1-NAA, IAA, 2,4-D (all Sigma-Aldrich) were kept as 20, 50, 50, 50, 10, 20, 80, 10, 20, 57,10 mM and 10, 10 and 10 mM DMSO stock solutions, respectively, and stored in −20 °C.

Jelínková A., Müller K., Fílová-Pařezová M, & Petrášek J. (2015). NtGNL1a ARF-GEF acts in endocytosis in tobacco cells. BMC Plant Biology, 15, 272.

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Visualizing endocytosis in N. benthamiana

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N. benthamiana leaves were infiltrated with 40 µM FM4-64 (Invitrogen) and then transferred to the microscope for imaging.
The endocytosis chemical inhibitors dynasore (Sigma) and Tyr A23 (Sigma) were used to inhibit the endocytic pathway, as previously reported (37 (link)– (link)39 (link)).

Wu G., Cui X., Chen H., Renaud J.B., Yu K., Chen X, & Wang A. (2018). Dynamin-Like Proteins of Endocytosis in Plants Are Coopted by Potyviruses To Enhance Virus Infection. Journal of Virology, 92(23), e01320-18.

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Inhibitor Infiltration in Transient Transformation

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All the drug treatments were performed by inhibitor infiltration in transiently transformed leaf. The final concentration of the inhibitor used were: 350 μM for tyrphostin A23 (TyrA23; Sigma-Aldrich²), 100 μM for salycid acid (SA; Sigma-Aldrich²), 2 μM for concanamycin A (ConA; Sigma-Aldrich²), 3 μM for wortmannin (Wm; Sigma-Aldrich²), 20 μM for Sortin 2 (ChemBridge Corporation³), 50 μM for Endosidin 5 (ES5; ChemBridge Corporation³).
To test BFA (Sigma-Aldrich²) effect on endocytosis it was necessary to distinguish between newly synthesized and endocytosed proteins, so leaves transiently expressing PGIP2-GFP and stably expressing secGFP-CesA6 were infiltrated with cycloheximide (CHX; Sigma-Aldrich²) to the final concentration of 300 μM. The same procedure was applied to ConA treatment on secGFP-CesA6 accordingly to Dettmer et al. (2006) (link). The leaf discs (1 cm diameter) taken from treated leaves were incubated by immersion in water containing 10 μM of FM4-64 dye (Invitrogen Molecular Probes⁴) before treatment with BFA. In the case of co-expression of secGFP-CesA6 with ST52-mCherry, CHX pre-treatment was not applied. Leaf discs were treated by immersion in solution with BFA at the final concentration of 100 μM and the induced fluorescent pattern was compared to the control conditions. Images were collected at the indicated times of incubation.

De Caroli M., Manno E., Perrotta C., De Lorenzo G., Di Sansebastiano G.P, & Piro G. (2020). CesA6 and PGIP2 Endocytosis Involves Different Subpopulations of TGN-Related Endosomes. Frontiers in Plant Science, 11, 350.

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VAEM Imaging of Epidermal Cells with Inhibitors

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TyrA23 and isobaxen were purchased from Sigma-Aldrich. The inhibitors were dissolved in 100% DMSO as stock solutions and further diluted in 1/2 MS for VAEM imaging on epidermal cells. The final concentration of DMSO was 0.1% or even lower in all working solutions. Four- to 5-d-old vertically grown seedlings were transferred from 1% agar plates to a well of a 12-well plate containing 4 mL of final working concentration in 1/2 MS. After the indicated incubation duration, seedlings were transferred to a glass slide with 100 μL inhibitor solution, covered with a glass coverslip for imaging as above.

Chen T., Ji D, & Tian S. (2018). Variable-angle epifluorescence microscopy characterizes protein dynamics in the vicinity of plasma membrane in plant cells. BMC Plant Biology, 18, 43.

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