Pepsin from porcine gastric mucosa (lyophilized powder, ≥3200 units/mg protein, lot # SLCH7086), α-ChymoTrypsin from bovine pancreas (lyophilized powder, ≥40 units/mg protein, lot # SLCH1926), Papain from papaya latex (≥8.0 units/mg protein, lot # SLCJ3270) and Trypsin from bovine pancreas (powder, ≥7500 BAEE units/mg solid, lot # SLCM7280) were purchased from Sigma-Aldrich (Oakville, ON, Canada). Here, papain was chosen since it comes from a vegetable to fully meet expectations of vegan or vegetarian people.
Trypsin from bovine pancreas
Manufactured by Merck Group
Sourced in United States, Germany
Trypsin from bovine pancreas is a proteolytic enzyme that catalyzes the hydrolysis of peptide bonds in proteins. It is commonly used in cell culture applications to dissociate adherent cells from the extracellular matrix and cell-cell contacts, facilitating subculturing and cell line maintenance.
Automatically generated - may contain errors
Lab products found in correlation
Pepsin from porcine gastric mucosa, by Merck Group (6 mentions)
Trypsin, by Merck Group (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
α chymotrypsin from bovine pancreas, by Merck Group (2 mentions)
Ethanolamine, by Merck Group (2 mentions)
Iodoacetamide iaa, by Merck Group (2 mentions)
Protein assay dye reagent concentrate, by Bio-Rad (2 mentions)
α amylase from bacillus licheniformis, by Merck Group (2 mentions)
Glycine, by Merck Group (2 mentions)
Chymotrypsin from bovine pancreas, by Merck Group (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Proteinase k from tritirachium album, by Merck Group (2 mentions)
Soybean trypsin inhibitor, by Merck Group (2 mentions)
Iodoacetamide, by Merck Group (2 mentions)
Peptide desalting spin column, by Thermo Fisher Scientific (2 mentions)
Speed vacuum concentrator savant sc210a, by Thermo Fisher Scientific (2 mentions)
Formic acid, by Merck Group (2 mentions)
Nupage bis tris gel, by Thermo Fisher Scientific (2 mentions)
Papain from papaya latex, by Merck Group (1 mentions)
59 protocols using trypsin from bovine pancreas
1
Vegan Enzyme Characterization Protocol
2
Immunohistochemical Analysis of Femur Cartilage
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemical analyses were done in 4 μm femur cartilage samples. Antigen retrieval was performed by incubation with 0.1% trypsin from bovine pancreas (Sigma-Aldrich, USA) in 0.1% CaCl2. Inflammatory markers were visualized using anti-metalloproteinase (MMP)-13 (R&D systems, USA, MAB511, 30 μg/ml) and anti-ciclooxigenase (COX)-2 (sc-1745; Santa Cruz Biotechnology, USA, 1/100 dilution) antibodies as previously described (30 (link)). A secondary biotinylated anti-mouse and anti-goat IgG was used respectively for detection of positive signal through a horseradish peroxidase linked to an avidin/biotin complex (ABC) (Vector Laboratories, USA) using 3,3 diaminobenzidine tetra-hydrochloride as chromogen (Dako, Denmark). Sections were counterstained with Haematoxylin, dehydrated and mounted in DPX (Merck Millipore, USA). Positive immunoreactivity was evaluated in x20 magnification photographs obtained using a Leica DM3000 LED digital micro-imaging instrument (Leica Microsystems, USA). Each image was analyzed using the Image J software (National Institutes of Health, USA) and the percentage of positive area was calculated as previously described (30 (link), 31 (link)).
3
Peptide Synthesis and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peptides were chemically synthesized by standard solid-phase methodology, and purity (>95%) and molecular weight were confirmed by reverse-phase chromatography and mass spectrometry, respectively. Tritrp–Dap was provided by PolyPeptide Group (San Diego, CA, USA) and all remaining peptides were manufactured by GenScript, Inc. (Piscataway, NJ, USA). Melittin was purified from honey bee venom (>70% purity) and was provided by Sigma Aldrich (St. Louis, MO, USA). The peptide concentrations were determined by absorbance at 280 nm, using the extinction coefficient provided by the ProtParam tool from the ExPASy server [47 ].
Cholesterol (Chol),l -α-phosphatidylcholine (ePC), and l -α-phosphatidylglycerol (ePG) from chicken eggs were provided as chloroform stocks by Avanti Polar Lipids, Inc. (Alabaster, AL, USA). Luria-Broth was purchased from BioShop Canada Inc. (Burlington, Canada). Trypsin from bovine pancreas and all other chemical reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA).
Escherichia coli ATCC 25922 was purchased from the American Type Culture Collection (Manassas, VA, USA) and E. coli ML35p was kindly provided by Dr. Robert Lehrer from the David Geffen School of Medicine at UCLA (Los Angeles, CA, USA).
Cholesterol (Chol),
Escherichia coli ATCC 25922 was purchased from the American Type Culture Collection (Manassas, VA, USA) and E. coli ML35p was kindly provided by Dr. Robert Lehrer from the David Geffen School of Medicine at UCLA (Los Angeles, CA, USA).
4
Aminopropyl Silica Characterization Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Aminopropyl silica (APS) (3 μm, 120 Å) was obtained from Osaka Soda (Amagasaki, Hyogo, Japan). Carboxymethyl-dextran sodium salt (CMD), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), N-hydroxysuccinimide (NHS), ethanolamine, sodium hydroxide pellets, potassium hydrogen phthalate (KHP), 2-(N-morpholino)ethanesulfonic acid (MES), phosphate buffered saline (PBS) pH 7.4 solution, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) liquid substrate (ABTS), horseradish peroxidase (HRP), trypsin from bovine pancreas (TPCK treated), bovine serum albumin (BSA), equine cytochrome c (Cyt c), human serum (H4522), Tris base, urea, iodoacetamide (IAA), dithiothreitol (DTT), Nα-benzoyl-l-arginine ethyl ester (BAEE) and formic acid 98% were purchased from Sigma Aldrich (Castle Hill, NSW, Australia). Anti-BSA monoclonal antibody (ab9092) was purchased from Abcam (Melbourne, Victoria, Australia). Mouse IgG VisUCyte HRP polymer antibody was obtained from R&D Systems (Minneapolis, MN, USA). Acetonitrile 99.9% purity (ACN) was obtained from Chem-Supply (Gillman, South Australia, Australia). Ultra-pure water (18.2 MΩ cm) obtained from a Sartorius 611 Arium® pro water generation system was used for all preparation and dilutions unless stated otherwise.
5
Trypsin-Mediated Cleavage of proCDTb
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For each reaction, 500 μg of proCDTb was mixed with Fab (BINTOXB/9, BINTOXN/19, or BINTOXB/22; 2-fold molar excess relative to proCTDb monomers) or buffer (for trypsinization in the absence of Fab) at a final concentration of 1 mg/mL proCDTb in 20 mM Tris (pH 8), 100 mM NaCl, 6 mM CaCl2. Trypsin from bovine pancreas (Sigma-Aldrich) was added at 1:20 (wt/wt) relative to proCDTb (25 μg) and the reaction mixture was incubated at 37°C for 20 min. The reaction was then run over a Superdex 200 Increase 10/300 GL (Cytiva) SEC column in 20 mM Tris (pH 8), 100 mM NaCl, 6 mM CaCl2.
6
Trypsin-Mediated Protein Digestion
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein samples (5 µg) were incubated with trypsin from bovine pancreas (0.002 µg/L; Sigma-Aldrich, Milan, Italy) at 20 °C in 50 mM NaHepes, pH 8.0, containing 300 mM NaCl and glycerol 5% (v/v). At specific time intervals, the proteolytic reaction was stopped by the addition of SDS and samples were boiled. Samples were then analyzed by SDS/PAGE.
7
Quantitative IGF-1 Measurement Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Ammonium bicarbonate (ABC), acetonitrile (HPLC purity), formic acid (analytical reagent grade), urea were purchased from Carlo Erba (Milan, Italy). Bovine insulin growth factor-1 (white powder, purity >95%), protein assay dye reagent concentrate were from Bio-Rad (Hercules, California, USA). Isotopically labeled IGF-1 peptide (LEMYCAPLKPA{K*} {K*} = Lys [13C6, 15 N2] isotopic enrichment: >99%) was from Pepscan (Presto AB Lelystad, NL). Bovine serum albumin (BSA), Tris-HCL, trypsin from bovine pancreas, iodoacetamide (IAA), dithiotreitol (DTT) were from Sigma-Aldrich® (Darmstadt, Germany).
8
Enzymatic Biofilm Disruption Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Enzyme treatments were performed on 4-day old biofilms. Planktonic cells were removed and biofilms were washed once with PBS prior to adding 150 μL of enzyme, or the corresponding buffer without enzyme as control. After 20 h of incubation at 37°C, planktonic cells were removed and the biofilm washed once with 150 μL PBS prior to quantification by crystal violet staining as described above. The following enzymes were used: TURBO® DNase (Thermo Fisher Scientific) in the commercial buffer provided, Cellulase from Trichoderma sp. (Sigma) in 50 mM citrate buffer (pH 5.0), α-amylase from Bacillus licheniformis (Sigma) in PBS (pH 7.4), α-mannosidase from Canavalia ensiformis (Sigma) in 10 mM ammonium acetate buffer (pH 7.0), Proteinase K (GoldBio) in 10 mM Tris buffer (pH 8.0), Trypsin from bovine pancreas (Sigma) in PBS (pH 7.4), Lipase from Candida rugosa (Sigma) in PBS (pH 7.4), Lysozyme (Sigma) in PBS (pH 7.4) and phospholipases A1 (from Aspergillus oryzae; Sigma) and A2 (from Apis mellifera; Sigma) in 50 mM Tris–HCl (pH 7.5), 10 mM CaCl2 and 2% DMSO buffer.
9
Cell Lines and Reagents for Cancer Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MDA-MB-231 and MCF-7 breast cancer cell lines were maintained in our institute, which were originally obtained from the American Type Culture Collection (ATCC). The U937 human promonocytic cell line was a gift from Professor Duan Ma (Key Laboratory of Molecular Medicine, Ministry of Education, Shanghai Medical College, Fudan University), which also originated from the ATCC. SD was purchased from the Pharmacy Department of our hospital, whose growth place was Inner Mongolia, China. Trypsin from bovine pancreas, dimethyl sulfoxide (DMSO), rhamnose (Rha), arabinose (Ara), xylose (Xyl), mannose (Man), glucose (Glc), and galactose (Gal) were purchased from Sigma (St. Louis, MO, USA). RPMI 1640 supplemented with 10% fetal bovine serum (FBS) and 100 units/mL penicillin/streptomycin was obtained from GIBCO (Invitrogen, Carlsbad, CA, USA). All other reagents used in this study were of analytical grade.
10
Enzymatic Modification of Native LDL
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A slight modification of the method described by Bhakdi et al. [30 (link)] was used for the generation of eLDL. Briefly, human native LDL (density 4 mg/mL) from plasma of healthy donors was isolated by ultracentrifugation [31 (link)]. LDL was diluted to 2 mg/mL in 20 mM HEPES, 150 mM NaCl, 2 mM CaCl2, pH 7.0. For enzymatic modification, LDL was digested with 4 µg/mL trypsin from bovine pancreas (Sigma-Aldrich) at 37 °C for 6 h and with 24 µg/mL cholesterol esterase from Pseudomonas sp. (Sigma-Aldrich) for an additional 6 h at 37 °C. Then, another 4 µg/mL trypsin and 36 µg/mL cholesterol esterase were added and the mixture was incubated at 37 °C for 24 h. Finally, trypsin activity was blocked by 10 µg/mL soybean trypsin inhibitor (Roche Diagnostics, Mannheim, Germany) for 60 min at 37 °C and modified LDL was dialyzed against PBS.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!