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Fk506

Manufactured by AG Scientific
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FK506 is an immunosuppressant drug used in pharmaceutical research and development. It functions as a calcineurin inhibitor, which affects the immune system's response. This product is primarily utilized in scientific investigations and studies, rather than for direct medical applications.

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Lab products found in correlation

Fluconazole, by LKT Laboratories (2 mentions) Nystatin, by Merck Group (2 mentions) Verapamil, by Merck Group (2 mentions) Valinomycin, by AG Scientific (2 mentions) Cyclosporine a, by AG Scientific (2 mentions) Doxorubicin, by AG Scientific (2 mentions) Fenpropimorph, by Merck Group (1 mentions) Aureobasidin a, by Takara Bio (1 mentions) Adenosine triphosphate (atp), by Merck Group (1 mentions) 5 foa, by US Biological (1 mentions) α ifnγ apc, by BD (1 mentions) Golgiplug protein transport inhibitor, by BD (1 mentions) αcd8 fitc, by BD (1 mentions) αcd3 pacificblue, by BD (1 mentions) Iotest beta mark, by Beckman Coulter (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Diva software, by BD (1 mentions)

8 protocols using fk506

1

Recombinant GST-FKBP12 Mutant Protocol

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Human recombinant glutathion-S-transferase (GST)-FKBP12 and its double mutant GST-FKBP12 (D37L, F99Y) were generous gifts from Dr. Yves Engelborghs, (Laboratory of Biomolecular Dynamics, K. U. Leuven, Belgium) [20 (link)]. Enzyme bovine G protein-coupled receptor kinase 2 (GRK2) was kindly provided by Dr. Jeffrey L. Benovic (Thomas Jefferson University, Philadelphia, PA) [21 (link)]. Morphine was supplied by the National Institute on Drug Abuse. FK506 was purchased from A.G. Scientific, Inc. (San Diego, CA, USA). All other chemicals were purchased from Sigma Chemical Co. (St. Louis, MO, USA).
Yan Y.H., Wang Y., Zhao L.X., Jiang S., Loh H.H., Law P.Y., Chen H.Z, & Qiu Y. (2014). Role of FK506 binding protein 12 in morphine-induced μ-opioid receptor internalization and desensitization. Neuroscience letters, 566, 231-235.
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2

Measuring Stress Response Pathways

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Strains harboring a 4xCDRE-lacZ construct were cultured in the absence or presence of 64 μg/ml fluconazole, and with or without 10 μM geldanamycin, 1 μg/ml FK506 (in DMSO, A. G. Scientific), or 20 μg/ml beauvericin. Strains were grown in synthetic defined (SD) media at 25°C overnight and diluted the next day to OD600 0.01 ± inhibitors geldanamycin, FK506, or beauvericin. Cultures ± inhibitors were again incubated overnight in SD at 25°C and diluted the next day to OD600 0.3 ± inhibitors in the presence or absence of fluconazole for five hours. Strains harboring the HSP70-lacZ construct were treated with 10 μM geldanamycin or 10 μg/ml beauvericin. Strains were grown in YEPD at 30°C overnight and diluted the next day to OD600 0.01 ± inhibitors geldanamycin or beauvericin. Cultures ± inhibitors were again incubated overnight in YEPD at 30°C and diluted the next day to OD600 0.2 ± inhibitors for four hours. Post-treatment sample processing and analysis was conducted as previously described10 (link).
Shekhar-Guturja T., Gunaherath G.M., Kithsiri Wijeratne E.M., Lambert J.P., Averette A.F., Lee S.C., Kim T., Bahn Y.S., Tripodi F., Ammar R., Döhl K., Niewola-Staszkowska K., Schmitt L., Loewith R.J., Roth F.P., Sanglard D., Andes D., Nislow C., Coccetti P., Gingras A.C., Heitman J., Leslie Gunatilaka A.A, & Cowen L.E. (2016). Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling. Nature chemical biology, 12(10), 867-875.
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3

Aspergillus niger Strain Characterization

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A. niger strains used in this study are listed in Table 3. The strains were grown at 30°C (unless otherwise stated) in minimal medium (MM) [61 ] or complete medium (CM), consisting of minimal medium (MM) supplemented with 1% yeast extract and 0.5% casamino acids. Fermentation medium (FM) was composed of 0.75% glucose, 0.45% NH4Cl, 0.15% KH2PO4, 0.05% KCl, 0.05% MgSO4, 0.1% salt solution [61 ] and 0.003% yeast extract. The pH of FM was adjusted to pH 3. Aureobasidin A was purchased from Takara Bio, FK506 from A.G. Scientific, fenpropimorph from Sigma Aldrich, caspofungin (Cancidas®) from Merck and calcofluor white from BASF.

Aspergillus nigerstrains used in this work

NameGenotypeReference
N402cspA1, amdS−[62 (link)]
MA169.4kusA::DR-amdS- DR, pyrG−[63 (link)]
MA78.6ΔkusA, pyrG+(derivative of MA70.15 containing A. niger pyrG)[63 (link)]
MA47.1ΔkusA, pyrG+, ΔrlmA ( derivative of AB4.1)[12 (link)]
ER2.5ΔkusA, ΔrhoB::AopyrG[13 (link)]
ER3.4ΔkusA, ΔrhoC::AopyrG[13 (link)]
ER7.6ΔkusA, ΔrhoD::AopyrG[13 (link)]
MA84.1ΔkusA, ΔcftA::hygR[13 (link)]
MA234.1pyrG+ (derivative of N402 containing A. niger pyrG)Unpublished
8.21ΔcrzA, pyrG+(derivative of MA234.1)Unpublished
MF3.2kusA::DR-amdS- DR, ΔrlmA, hphThis study
JH1.1kusA::DR-amdS-DR, msnA−, hphThis study
MF4.10kusA::DR-amdS- DR, msnA−, ΔrlmA, hphThis study
Fiedler M.R., Lorenz A., Nitsche B.M., van den Hondel C.A., Ram A.F, & Meyer V. (2014). The capacity of Aspergillus niger to sense and respond to cell wall stress requires at least three transcription factors: RlmA, MsnA and CrzA. Fungal Biology and Biotechnology, 1, 5.
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4

Measuring Stress Response Pathways

Cited 1 time
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Strains harboring a 4xCDRE-lacZ construct were cultured in the absence or presence of 64 μg/ml fluconazole, and with or without 10 μM geldanamycin, 1 μg/ml FK506 (in DMSO, A. G. Scientific), or 20 μg/ml beauvericin. Strains were grown in synthetic defined (SD) media at 25°C overnight and diluted the next day to OD600 0.01 ± inhibitors geldanamycin, FK506, or beauvericin. Cultures ± inhibitors were again incubated overnight in SD at 25°C and diluted the next day to OD600 0.3 ± inhibitors in the presence or absence of fluconazole for five hours. Strains harboring the HSP70-lacZ construct were treated with 10 μM geldanamycin or 10 μg/ml beauvericin. Strains were grown in YEPD at 30°C overnight and diluted the next day to OD600 0.01 ± inhibitors geldanamycin or beauvericin. Cultures ± inhibitors were again incubated overnight in YEPD at 30°C and diluted the next day to OD600 0.2 ± inhibitors for four hours. Post-treatment sample processing and analysis was conducted as previously described10 (link).
Shekhar-Guturja T., Gunaherath G.M., Kithsiri Wijeratne E.M., Lambert J.P., Averette A.F., Lee S.C., Kim T., Bahn Y.S., Tripodi F., Ammar R., Döhl K., Niewola-Staszkowska K., Schmitt L., Loewith R.J., Roth F.P., Sanglard D., Andes D., Nislow C., Coccetti P., Gingras A.C., Heitman J., Leslie Gunatilaka A.A, & Cowen L.E. (2016). Dual Action Antifungal Small Molecule Modulates Multidrug Efflux and TOR Signaling. Nature chemical biology, 12(10), 867-875.
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5

Membrane Protein Assay Reagents

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FK506, cyclosporine A, doxorubicin and valinomycin were purchased from A.G. Scientific. Fluconazole was from LKT Laboratories. Verapamil, nystatin and ATP were from Sigma Aldrich. Escherichia coli lipids (Polar Extract) were purchased from Avanti Polar Lipids, and DDM (n-dodecyl-β-D-maltopyranoside) was from Inalco. CP-MTS [7-diethylamino-3-(4′-maleimidylphenyl)-4-methylcoumarin] was from Biotium.
Swartz D.J., Mok L., Botta S.K., Singh A., Altenberg G.A, & Urbatsch I.L. (2014). Directed evolution of P-glycoprotein cysteines reveals site-specific, non-conservative substitutions that preserve multidrug resistance. Bioscience Reports, 34(3), e00116.
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6

Multidrug Screening in Yeast Model

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FK506, cyclosporine A, doxorubicin, and valinomycin were purchased from A.G. Scientific (San Diego, CA). Fluconazole was from LKT Laboratories (Saint Paul, MN). Verapamil, and nystatin were from Sigma Aldrich (Saint Louis, MO). 5-FOA and G418 were from US Biological (Swampscott, Massachusetts). E. coli lipids (Polar Extract) was purchased from Avanti (Alabaster, AL), n-dodecyl-β-D-maltopyranoside (DDM) was from Inalco (Italy).
Swartz D.J., Singh A., Sok N., Thomas J.N., Weber J, & Urbatsch I.L. (2020). Replacing the eleven native tryptophans by directed evolution produces an active P-glycoprotein with site-specific, non-conservative substitutions. Scientific Reports, 10, 3224.
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7

EBV Infection of PBMC with FK506

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PBMC from healthy subjects were infected with EBV (wild type p2089 virus or mutated for SZF1-binding sites) at MOI of 1 in the presence of 20nM FK506. Cells were incubated with FK506 (AG Scientific) for an hour at 37° C before infection. Infected cells were left in culture to establish LCL.
Burton E.M., Akinyemi I.A., Frey T.R., Xu H., Li X., Su L.J., Zhi J., McIntosh M.T, & Bhaduri-McIntosh S. (2021). A heterochromatin inducing protein differentially recognizes self versus foreign genomes. PLoS Pathogens, 17(3), e1009447.
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8

Expansion and Characterization of HIV-Specific CD8+ T Cells

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B-EBV lines were generated for each donor by culturing PBMCs in RPMI, 20% fetal bovine serum (FBS), 20nM FK506 (AG Scientific), and Ebstein-barr virus (EBV)-containing supernatant from the virus-producing B95.8 marmoset cell line (ATCC) at an MOI of 100. B-EBV cell lines were then loaded with 0.5–5 mg/mL HIV Clade AE peptide pools overnight. Peptide pools were made with 20 15-mer peptides per pool of HIV PTE and HIV Consensus A peptides (gag, pol, nef, env) obtained through the AIDS Reagent Program, Division of AIDS, NIAID. Expanded primary CD8+ T cells were added to the loaded B-EBV (2:1), co-stimulated with 1 μg/μL αCD28/CD49d (BD Biosciences) and incubated for 12 hours with GolgiPlug protein transport inhibitor (BD Biosciences). After incubation, cells were stained with αCD8-FITC, αCD3-PacificBlue (BD Biosciences), and αCD20-PECy7 (BioLegend) prior to fixation/permeabilization and intracellular staining with αIFNγ-APC (BD Biosciences). Live/dead stain with Vivid-amcyan was used to exclude dead cells from the analysis. The T-cell receptor Vbeta repertoire was analyzed by flow cytometry using the IOTest® Beta mark (Beckman Coulter) in conjunction with αCD8-Pacific Blue (BD Biosciences). Stained CD8+ T cells were run on an LSRII flow cytometer using DiVA software (BD Biosciences) and analyzed with FlowJo (Treestar).
Kessing C.F., Spudich S., Valcour V., Cartwright P., Chalermchai T., Fletcher J.L., Nichols C., Josey B.J., Slike B., Krebs S.J., Sailsuta N., Lerdlum S., Jagodzinski L., Tipsuk S., Suttichom D., Rattanamanee S., Zetterberg H., Hellmuth J., Phanuphak N., Robb M.L., Michael N.L., Ananworanich J, & Trautmann L. (2017). High Number of Activated CD8+ T Cells Targeting HIV Antigens are Present in Cerebrospinal Fluid in Acute HIV Infection. Journal of acquired immune deficiency syndromes (1999), 75(1), 108-117.
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