Pepstatin

Manufactured by GoldBio

Pepstatin is a protease inhibitor that inhibits aspartic proteases, such as pepsin, renin, and certain cathepsins. It is a cyclic pentapeptide with the amino acid sequence Isovaleryl-Val-Val-Sta-Ala-Sta, where Sta represents statine, a nonproteinogenic amino acid. Pepstatin is commonly used in research applications to prevent proteolytic degradation of proteins.

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Lab products found in correlation

Aprotinin, by GoldBio (2 mentions) Leupeptin, by GoldBio (2 mentions) Fastdigest restriction enzyme, by Thermo Fisher Scientific (1 mentions) Anti mouse irdye 680rd, by LI COR (1 mentions) Nanoglow luciferase assay, by Promega (1 mentions) E coli krx cells, by Promega (1 mentions) Anti rabbit irdye 800cw, by LI COR (1 mentions) Nanobit, by Promega (1 mentions) Pfu ultra dna polymerase, by Agilent Technologies (1 mentions) γ 32p gtp, by PerkinElmer (1 mentions) F1804, by Merck Group (1 mentions)

2 protocols using pepstatin

Peptide Synthesis and Epigenetic Assays

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All cloning reagents were obtained from New England Biolab (Gibson assembly Master Mix, Q5-mutagenesis kit) or Thermo Scientific (Fast Digest restriction enzymes). Reagents and building blocks for the solid phase peptide synthesis (SPPS) were from Anaspec, P3Bio, ChemPep, or Aaptec. The Chem Matrix Rink Amide resin was from Sigma. Solvents for SPPS, HPLC, and LC/MS, buffering agents, and additives were from Sigma or Fisher Scientific. Antibiotics, IPTG, DTT, and protease inhibitors (leupeptin, pepstatin, aprotinin, AEBSF) were from Gold Bio. KRX E.coli cells, NanoBit, and NanoGlow luciferase assay components were from Promega, and L-rhamnose was from Chem-Implex. tRNA from yeast was purchased from Sigma. Corning 384 Well Low Volume Black Round Bottom Polystyrene NBS Microplates were from Sigma. Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (ab1191) was from Abcam, Anti-Histone H3 (acetyl K9) antibody (ab4441) was from Abcam. Secondary antibodies IRDye 680RD anti-mouse and IRDye 800CW anti-rabbit were from Li-Cor.

Kuznetsov V.I., Liu W.H., Klein M.A, & Denu J.M. (2022). Potent activation of NAD+-dependent deacetylase Sirt7 by nucleosome binding. ACS chemical biology, 17(8), 2248-2261.

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Recombinant Protein Expression and Purification

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Unless otherwise indicated, all chemical reagents were obtained from Sigma or Fisher Scientific. Fluorescein di-β-D-galactopyranoside (FDG) was from Marker Gene Technologies, and the protein inhibitor mixture was from Sigma (catalog no. S8830). Leupeptin, pepstatin, and aprotinin were from Gold Biotechnology. All restriction endonucleases and E. coli strain BL21(DE3) were from Thermo Scientific. E. coli strain DH5α was purchased from New England Biolabs. Pfu ultra DNA polymerase used for site-directed mutagenesis was purchased from Agilent. [γ-³²P]GTP was from Perkin Elmer. Mouse monoclonal antibodies raised against α-tubulin (T6074), FLAG tag (F1804) or His-tag (H1029) were from Sigma. Mouse monoclonal antibody raised against hemagglutinin (HA) tag (clone 12CA5, #11583816001) was obtained from Roche. Mouse monoclonal antibody raised against MYC-tag (9B11, #2276) was from Cell Signaling. Rabbit polyclonal antibody raised against Gαs (C-18, sc-383) was purchased from Santa Cruz Biotechnology. Goat anti-rabbit Alexa Fluor 680 (A21077) and goat anti-mouse IRDye 800 (#926–32210) secondary antibodies were from Life technologies and LiCor, respectively.

Garcia-Marcos M., Parag-Sharma K., Marivin A., Maziarz M., Luebbers A, & Nguyen L.T. (2020). Optogenetic activation of heterotrimeric G-proteins by LOV2GIVe, a rationally engineered modular protein. eLife, 9, e60155.

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