Pfi 3

Manufactured by Cayman Chemical

Sourced in United States

PFI-3 is a compact and versatile laboratory instrument designed for the purification and isolation of proteins, peptides, and other biomolecules. It utilizes fast protein liquid chromatography (FPLC) technology to provide high-resolution separation and purification. The PFI-3 is suitable for a wide range of applications in biochemistry, molecular biology, and proteomics research.

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Lab products found in correlation

Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (2 mentions) Simplechip chromatin ip buffers, by Cell Signaling Technology (2 mentions) Vectastain elite abc pk 6200, by Vector Laboratories (2 mentions) Mneongreen, by Agilent Technologies (2 mentions) Calpain inhibitor 3, by Cayman Chemical (2 mentions) Gsk602, by Selleck Chemicals (1 mentions) Sc 7963, by Santa Cruz Biotechnology (1 mentions) Sc 17796, by Santa Cruz Biotechnology (1 mentions) Chir99021, by Cayman Chemical (1 mentions) Mayer s hematoxylin, by Merck Group (1 mentions) Optimal cutting temperature compound, by Thermo Fisher Scientific (1 mentions) Eosin y, by Merck Group (1 mentions) Corn oil, by Merck Group (1 mentions) Anti β tubulin, by Santa Cruz Biotechnology (1 mentions) Xylene, by Electron Microscopy Sciences (1 mentions) Anti vinculin, by Santa Cruz Biotechnology (1 mentions) Myogenin f5d, by Developmental Studies Hybridoma Bank (1 mentions)

5 protocols using pfi 3

Synthesis and Characterization of Novel Epigenetic Inhibitors

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N′‐[1,1‐bis(oxidanylidene)thian‐4‐yl]‐5‐ethyl‐4‐oxidanylidene‐7‐[3‐(trifluoromethyl)phenyl]thieno[3,2‐c]pyridine‐2‐carboximidamide was purchased from Selleck Chemicals (PubChem CID 91668541, iBRD9, I‐BRD9, GSK602, Houston, Texas, USA). 4‐[4‐[(dimethylamino)methyl]‐3,5‐dimethoxy‐phenyl]‐2‐methyl‐2,7‐naphthyridin‐1‐one (PubChem CID 118796358, BI7273) and (E)‐1‐(2‐hydroxyphenyl)‐3‐((1R,4R)‐5‐(pyridin‐2‐yl)‐2,5‐diazabicyclo[2.2.1]heptan‐2‐yl)prop‐2‐en‐1‐one (PubChem CID 78243717, PFI3, PFI‐3) were purchased from Cayman Chemicals.

Basuroy T., Dreier M., Baum C., Blomquist T., Trumbly R., Filipp F.V, & de la Serna I.L. (2022). Epigenetic and pharmacological control of pigmentation via Bromodomain Protein 9 (BRD9). Pigment Cell & Melanoma Research, 36(1), 19-32.

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Chromatin Immunoprecipitation and Western Blotting Protocol

Cited 1 time

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Primary antibodies used for WB against BRG1 (sc-17796; 1:1,000), LaminB1 (sc-56144; 1:10,000) and β-catenin (sc-7963, 1:1,000) were purchased from Santa Cruz Biotech, United States. Myosin Heavy Chain (MHC) (Catalog# MF20; 1:1,000 for WB; 1:100 for ICC) was purchased from the Developmental Studies Hybridoma Bank, University of Iowa, United States. Antibodies used for chromatin IPs against β-catenin (Cell Signaling Technology, Inc., United States, Catalog# 8,480, 2 μL per μg of chromatin), BRG1 (Santa Cruz Biotech, United States, sc-17796, G-7X, 3–4 μL per μg of chromatin) and BRM antisera (3–4 μL per μg of chromatin) have been previously described (de la Serna et al., 2005 (link); Sharma et al., 2021 (link)). PFI-3 (catalog# 15267) and CHIR99021 (catalog# 13122) were purchased from Cayman Chemicals, United States. Lysis buffers for ChIP assays were purchased from Cell Signaling Technology, United States (SimpleChIP^® Enzymatic Cell Lysis Buffers A & B, 14,282; SimpleChIP^® Chromatin IP Buffers, 14,231). Dulbecco’s modified Eagle’s medium (DMEM) was purchased from ThermoFisher Scientific (#11965118). Vectastain elite ABC (PK-6200) and HRP DAB substrate (SK-4100) kits were purchased from Vector Laboratories, United States.

Sharma T., Olea-Flores M, & Imbalzano A.N. (2023). Regulation of the Wnt signaling pathway during myogenesis by the mammalian SWI/SNF ATPase BRG1. Frontiers in Cell and Developmental Biology, 11, 1160227.

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High-throughput SARS-CoV-2 Antiviral Screening

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Calpain Inhibitor III (#14283), SIS3 (#15495), APY0201 (#9001589), and PFI-3 (#15267) were purchased from Cayman Chemical. Drugs were resuspended at a stock concentration of 40mM in DMSO and then two-fold serial dilutions were performed in DMSO. 20 nanoliters of 1000X drug stock were spotted into each well of a 384-well plate using Labcyte ECHO acoustic dispenser at the Yale Center for Molecular Discovery. 1.25 × 10³ Vero-E6 cells were plated per well in 20 μl of phenol-red free DMEM containing 5% FBS. Two days later, 5,000 PFU (MOI ~1) icSARS-CoV-2-mNG in 5 μl media was added. Cells were incubated at 37°C and 5% CO2 for two days. Infected cell frequencies were quantified by mNeonGreen at 2 dpi (Cytation 5, BioTek). In parallel, 500 PFU (MOI~0.2) SARS-CoV-2 in 5 μl media was added to replicate plates and cell viability was quantified by CellTiter Glo at 3 dpi.

Wei J., Alfajaro M.M., Hanna R.E., DeWeirdt P.C., Strine M.S., Lu-Culligan W.J., Zhang S.M., Graziano V.R., Schmitz C.O., Chen J.S., Mankowski M.C., Filler R.B., Gasque V., de Miguel F., Chen H., Oguntuyo K., Abriola L., Surovtseva Y.V., Orchard R.C., Lee B., Lindenbach B., Politi K., van Dijk D., Simon M.D., Yan Q., Doench J.G, & Wilen C.B. (2020). Genome-wide CRISPR screen reveals host genes that regulate SARS-CoV-2 infection. bioRxiv.

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SARS-CoV-2 Infection Inhibition Assay

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Calpain Inhibitor III (#14283), SIS3 (#15495), and PFI-3 (#15267) were purchased from Cayman Chemical. Drugs were resuspended at a stock concentration of 40 mM in DMSO and then two-fold serial dilutions were performed in DMSO. 20 nL of 1000X drug stock were spotted into each well of a 384-well plate using Labcyte ECHO acoustic dispenser at the Yale Center for Molecular Discovery. 1.25 × 10³ Vero-E6 cells were plated per well in 20 μl of phenol-red free DMEM containing 5% FBS. Two days later, 5,000 PFU (MOI ∼1) icSARS-CoV-2-mNG in 5 μl media was added. Cells were incubated at 37°C and 5% CO2 for two days. Infected cell frequencies were quantified by mNeonGreen at 2 dpi (Cytation 5, BioTek). In parallel, 1000 PFU (MOI∼0.2) SARS-CoV-2 in 5 μl media was added to replicate plates and cell viability was quantified by CellTiter Glo at 3 dpi. Vero-E6, Huh7.5 and Calu-3 cells were pretreated with 10 μM SIS3 and 40 μM PFI-3 for 48 hours and then infected with SARS-CoV-2 at a MOI of 0.1. Viral production was determined by plaque assay. Cytotoxicity was not observed in these cell lines during the time and concentration of drug used.

Wei J., Alfajaro M.M., DeWeirdt P.C., Hanna R.E., Lu-Culligan W.J., Cai W.L., Strine M.S., Zhang S.M., Graziano V.R., Schmitz C.O., Chen J.S., Mankowski M.C., Filler R.B., Ravindra N.G., Gasque V., de Miguel F.J., Patil A., Chen H., Oguntuyo K.Y., Abriola L., Surovtseva Y.V., Orchard R.C., Lee B., Lindenbach B.D., Politi K., van Dijk D., Kadoch C., Simon M.D., Yan Q., Doench J.G, & Wilen C.B. (2020). Genome-wide CRISPR Screens Reveal Host Factors Critical for SARS-CoV-2 Infection. Cell, 184(1), 76-91.e13.

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Chromatin Immunoprecipitation and Myogenesis Assays

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Antibodies were purchased from ABclonal Technology, USA (anti-PBRM1, A9878) and Santa Cruz Biotech, USA (anti-BRG1, sc-17796; anti-BrdU, sc-32323; anti-Cyclin A, sc-596; anti-Cyclin E, sc-198; anti-Vinculin, sc-25336; anti-LaminB1, sc-56144; anti-β-tubulin, sc-166729). Anti-acetyl histone H3 (06–599) and anti-acetyl histone H4 (06–598) were purchased from Miilipore Sigma, USA. Myosin Heavy Chain (MHC) (#MF20) and Myogenin (#F5D) antibodies were purchased from the Developmental Studies Hybridoma Bank, University of Iowa, USA. BRM and Myod antisera were described previously (25 (link),58 (link)). PFI-3 was purchased from Cayman Chemicals, USA. Lysis buffers for ChIP assays were purchased from Cell Signaling Technology, USA (SimpleChIP^® Enzymatic Cell Lysis Buffers A & B, 14282; SimpleChIP^® Chromatin IP Buffers, 14231). Dulbecco's modified Eagle's medium (DMEM) was purchased from ThermoFisher Scientific (#11965118). Vectastain elite ABC (PK-6200) and HRP DAB substrate (SK-4100) kits were purchased from Vector Laboratories, USA. Corn Oil, Mayer's hematoxylin, Eosin Y and lithium carbonate were purchased from Millipore Sigma, USA. Cardiotoxin was purchased from Laxtoxan, France. Xylene was purchased from Electron Microscopy sciences, USA and Optimal Cutting Temperature (OCT) compound was from Fisher Healthcare, USA.

Sharma T., Robinson D.C., Witwicka H., Dilworth F.J, & Imbalzano A.N. (2021). The Bromodomains of the mammalian SWI/SNF (mSWI/SNF) ATPases Brahma (BRM) and Brahma Related Gene 1 (BRG1) promote chromatin interaction and are critical for skeletal muscle differentiation. Nucleic Acids Research, 49(14), 8060-8077.

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