Papain

Manufactured by MP Biomedicals

Sourced in United States

Papain is a proteolytic enzyme derived from the papaya plant. It is a versatile lab equipment used for various applications in research and industrial settings. Papain possesses the ability to break down protein molecules through hydrolysis.

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Lab products found in correlation

L cysteine, by MP Biomedicals (2 mentions) Trans 4 hydroxy l proline, by Thermo Fisher Scientific (1 mentions) Quant it picogreen kit, by Thermo Fisher Scientific (1 mentions) Carbazole, by Merck Group (1 mentions) Acetic acid, by Thermo Fisher Scientific (1 mentions) Streptavidin cy3, by Thermo Fisher Scientific (1 mentions) Hyaluronic acid sodium salt from streptococcus equi, by Merck Group (1 mentions) Cell counting kit 8 cck 8, by Dojindo Laboratories (1 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions) Powerup sybr green master mix, by Thermo Fisher Scientific (1 mentions) N acetylcysteine, by Merck Group (1 mentions) Sodium tetraborate, by Merck Group (1 mentions) Rneasy plant mini kit, by Qiagen (1 mentions) Yeast extract, by Hardy Diagnostics (1 mentions) Todd hewitt medium, by HiMedia (1 mentions) Disodium edta, by Merck Group (1 mentions) Synergy ht, by Agilent Technologies (1 mentions) Sodium acetate, by Merck Group (1 mentions) Terpineol, by Merck Group (1 mentions) Salicylic acid, by Merck Group (1 mentions)

5 protocols using papain

Quantitative Analysis of Extracellular Matrix Components

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Compositional analysis was performed as per previous reports [39 (link)]. Briefly, constructs were lyophilized at –55 °C and 0.2 mbar for 4–5 h and then 20 mg of each sample was digested (9 mM di-sodium EDTA, 20 mM sodium acetate, 20 mM l-cysteine (MP Biomedicals, Solon, OH, USA), 2 mg papain (MP Biomedicals) per gram lyophilized sample) at 60 °C for 10 h [40 (link), 41 (link)]. Digested samples were vigorously vortexed and then centrifuged (4000 × g, 10 min). Supernatants were stored at –80 °C until analysis. Double-stranded DNA was determined with a commercially available kit (Quant-iT™ PicoGreen® Kit, Invitrogen, Carlsbad, CA, USA) [41 (link)]. Hydroxyproline content as a measure of total collagen was quantified via Ehrlich’s colorimetric assay and absorbance at 550 nm based on trans-4-hydroxy-l-proline (ACROS Organics™, Morris Plains, NJ, USA) standards [41 (link), 42 (link)]. Sulfated glycosaminoglycan (sGAG) was quantified with a dimethylmethylene blue (DMMB) assay, with absorbance at 520 nm and a chondroitin sulfate standard curve [41 (link), 43 (link)]. Lowry’s total protein assay with Biuret’s and Folin-Ciocalteu’s reagents, absorbance at 650 nm, and bovine serum albumin standards, was used to quantify sample protein [44 (link)].

Duan W., Chen C., Haque M., Hayes D, & Lopez M.J. (2018). Polymer-mineral scaffold augments in vivo equine multipotent stromal cell osteogenesis. Stem Cell Research & Therapy, 9, 60.

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Extracellular Matrix Components Characterization

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Fibrillar collagen type I was obtained from Collagen Matrix (Oakland, NJ). Chondroitin sulfate (CS) from shark cartilage, hyaluronic acid (HA) sodium salt from Streptococcus equi, n-acetyl cysteine, EthylenediaminetetraAcetic acid (EDTA), sodium phosphate, carbazole, and sodium tetraborate were obtained from Sigma Aldrich (St. Louis, MO). Acetic acid, low glucose Dulbecco’s Minimum Essential Medium (DMEM-LG), and streptavidin-CY3 were obtained from Invitrogen (Carlsbad, CA). Cell Counting kit-8 (CCK8) was obtained from Dojindo Molecular Technologies (Rockville, MD). Anti-Fibronectin antibody (Biotin) (ab6584) was obtained from Abcam (Cambridge, UK). Goat anti-rabbit IgG (E0432) was obtained from Dako (Denmark). RNeasy Plant Mini Kit was obtained from Qiagen (Hilden, Germany). High-Capacity cDNA Reverse Transcription kit and PowerUp SYBR Green master mix were obtained from Applied Biosystems (Foster City, CA). Papain was obtained from MP Biomedicals (Solon, OH). Human trabecular meshwork (hTM) cells (Cat# 7278) from a 20 week old female donor were purchased from Sciencell Research Laboratories (Carlsbad, CA). These cells were confirmed by us to be trabecular meshwork cells based on measuring a robust increase in myocilin expression after treatment with dexamethasone.(Keller et al., 2018 (link))

Osmond M.J., Krebs M.D, & Pantcheva M.B. (2020). Human Trabecular Meshwork Cell Behavior is Influenced by Collagen Scaffold Pore Architecture and Glycosaminoglycan Composition. Biotechnology and bioengineering, 117(10), 3150-3159.

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Purification of Streptococcus pyogenes Zymogen SpeB

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S. pyogenes 10782 zymogen SpeB clone (residues 28–398) was overexpressed and purified as a C-terminal His₆-tag fusion from E. coli BL21DE3 (Strategene) in a pET23b vector (Novagen), as previously described.^{21 (link), 42 (link)} Caspase-3 was expressed and purified as previously described.⁴³ Papain was purchased from MP Biomedicals, reconstituted, and purified by size-exclusion chromatography. Human sputum leucocyte elastase was purchased from Elastin Products Company, Inc. and human complement C3 and C3b were purchased from Complement Technologies. Streptococcus pyogenes serotype M1 (strain 5448), WT or ΔspeB, was grown in Todd-Hewitt medium (HiMedia Laboratories) supplemented with 0.2% yeast extract (Difco) (THY media) or in C medium which consists of 0.5% Proteose Peptone #3 (Hardy Diagnostics), 1.5% yeast extract, 10 mM K₂HPO₄, 0.4 mM MgSO₄, 17 mM NaCl, adjusted to pH 7.5.^{44 (link)}

Woehl J.L., Kitamura S., Dillon N., Han Z., Edgar L.J., Nizet V, & Wolan D.W. (2020). An irreversible inhibitor to probe the role of Streptococcus pyogenes cysteine protease SpeB in evasion of host complement defenses. ACS chemical biology, 15(8), 2060-2069.

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Quantitative Compositional Analysis of Cell-Scaffold Constructs

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Cell–scaffold constructs were lyophilized (−55°C, 0.2 mbar, 4 h) and then incubated in a papain digest (2 mg papain [MP Biomedicals]/g lyophilized sample, 20 mM L-cysteine [MP Biomedicals], 9 mM disodium EDTA [Sigma-Aldrich], and 3.6 M sodium acetate [Sigma-Aldrich]) at 60°C for 10 h. Following centrifugation of the solution (4000 g, 10 min), the supernatant was collected and stored at −80°C. Samples were thawed for 1 min in a 37°C water bath immediately before compositional analysis. All assays were performed according to manufacturer's instructions or published methods for microtiter plate analysis and fluorescence or absorbance quantified with a multiwell plate reader (Synergy HT; BioTek Instruments, Winooski, VT). Composition fold change relative to day 0 was calculated as C_f/C_i (C_i: content immediately after cell loading; C_f: content after 7, 14, or 28 days of culture).

Duan W., Haque M., Kearney M.T, & Lopez M.J. (2017). Collagen and Hydroxyapatite Scaffolds Activate Distinct Osteogenesis Signaling Pathways in Adult Adipose-Derived Multipotent Stromal Cells. Tissue Engineering. Part C, Methods, 23(10), 592-603.

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Transungual Drug Delivery Formulations

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Econazole Nitrate (≥98%) was purchased from Cayman Chemical Company (Ann Arbor, Michigan, USA). Eudragit polymers and Hydroxypropyl Methylcellulose (HPMC) were kind gifts from Evonik Corporation (Piscataway, New Jersey, USA) and Ashland (Covington, Kentucky), respectively. Transcutol P was a gift from Gattefosse (Paramus, New Jersey, USA). Kolliphor CS 20 and Kolliphor RH 40 were generous gifts from BASF (Tarrytown, New York, USA). Isopropyl Alcohol (IPA), Acetone, Ethanol (200 proof), HPLC grade water, Tween 80, Terpineol, Polyethylene Glycol 400 and Salicylic Acid were purchased from Sigma-Aldrich, Inc. (St. Louis, Missouri, USA). Triacetin was purchased form TCI Chemicals (Portland, Oregon, USA), Papain was purchased from MP Biomedicals, LLC (Irvine, California, USA) and phosphate-buffered saline (PBS) was purchased in the form of tablets from Tocris, Bio-Techne Corporation (Minneapolis, Minnesota, USA). Dexpanthenol was purchased from Spectrum Chemical Mfg. Corp. (New Brunswick, New Jersey, USA), Urea was purchased from Santa Cruz Biotechnology (Dallas, TX). Human cadaver fingernail samples were purchased from Science Care (Phoenix, Arizona, USA). (Note: Cadaver tissue samples were obtained from an accredited Tissue Bank and could not be linked to specific individuals and, therefore, did not require approval from an Ethics Committee or Institutional Review Board.)

Puri V., Savla R., Chen K., Robinson K., Virani A, & Michniak-Kohn B. (2022). Antifungal Nail Lacquer for Enhanced Transungual Delivery of Econazole Nitrate. Pharmaceutics, 14(10), 2204.

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