Chymotrypsin

Manufactured by Promega

Sourced in United States, Australia, Italy, United Kingdom

Chymotrypsin is a proteolytic enzyme derived from bovine pancreas. It is commonly used in biochemical and molecular biology applications to cleave peptide bonds following aromatic amino acid residues, such as tyrosine, tryptophan, and phenylalanine.

Automatically generated - may contain errors

Lab products found in correlation

Trypsin, by Promega (42 mentions) Sequencing grade trypsin, by Promega (14 mentions) Iodoacetamide, by Merck Group (12 mentions) Glu c, by Promega (12 mentions) Asp n, by Promega (11 mentions) Ammonium bicarbonate, by Merck Group (9 mentions) Formic acid, by Thermo Fisher Scientific (7 mentions) Dithiothreitol (dtt), by Merck Group (6 mentions) Lys c, by Promega (6 mentions) Urea, by Merck Group (5 mentions) Trypsin lys c, by Promega (4 mentions) Pepsin, by Promega (4 mentions) Pngase f, by New England Biolabs (3 mentions) 4 vinylpyridine, by Merck Group (3 mentions) Optima lc ms grade acetonitrile, by Thermo Fisher Scientific (3 mentions) Trypsin gold, by Promega (3 mentions) Iodoacetamide iaa, by Merck Group (3 mentions) Sequencing grade modified trypsin, by Promega (3 mentions) Trypsin lys c mix, by Promega (3 mentions) Trizma base, by Merck Group (3 mentions)

Close spelling variants of this product

Proteasome-Glo Chymotrypsin-Like Cell-Based Assay (23 citations) Sequencing grade chymotrypsin (11 citations) Proteasome-Glo Chymotrypsin-Like Assay (6 citations) Proteasome-Glo Chymotrypsin-like Cell-Based Assay kit (6 citations) Proteasome-Glo Chymotrypsin-Like Assay kit (4 citations) Cell-Based Proteasome-Glo Chymotrypsin-like kit (4 citations) Proteasome-Glo™ Chymotrypsin-Like Cell-Based Assay Reagent (3 citations) Proteasome-Glo™ Chymotrypsin-Like (3 citations) α-chymotrypsin (3 citations) Chymotrypsin (Mass Spectrometry Grade, (3 citations)

100 protocols using chymotrypsin

Proteome Exploration via In-Gel Digestion

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The protocol has been described previously^{57 (link)}. Briefly, SDS-PAGE bands were excised from the gel and transferred into 96-well microtitration plates. The bands were then destained and dehydrated. Gel pieces were washed again with the destaining solutions and then incubated with trypsin (Promega, Madison, WI) and chymotrypsin (Promega) for digestion overnight at room temperature. The resulting peptides were extracted from the gel pieces. The initial digestion and extraction supernatants were pooled together and vacuum-dried in a SpeedVac concentrator.

des Georges A., Dhote V., Kuhn L., Hellen C.U., Pestova T.V., Frank J, & Hashem Y. (2015). Structure of mammalian eIF3 in the context of the 43S preinitiation complex. Nature, 525(7570), 491-495.

+ Open protocol

+ Expand

Detecting and Characterizing Protein Unfolding

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tris(2-carboxyethyl)phosphine (TCEP) was obtained from Thermo Scientific (Rockford, IL). Iodoacetamide, tunicamycin, glucosamine, and mouse monoclonal anti-FLAG M2 antibody were obtained from Sigma-Aldrich (St. Louis, MO). Rabbit polyclonal anti-human GntI/MGAT1 antibody (15103-1-AP) was obtained from Proteintech. Mouse monoclonal anti-CHOP (9C8; MA1-250) was obtained from Thermo Fisher Scientific. Rabbit anti-human/mouse XBP-1 antibody was obtained from Abcam (ab37152). Rabbit monoclonal antibodies against spliced XBP-1s (D2C1F) and ATF-6 (D4Z8 V) were obtained from Cell Signaling Technology. Trypsin and chymoTrypsin were purchased from Promega (Madison, WI); hydrazine resin was from Bio-Rad (Hercules, CA), and PNGase F and endoglycosidase H (Endo H) were from New England Biolabs (Ipswich, MA). The furin/PCSK3 inhibitor (decanoyl-Arg-Val-Lys-Arg-CMK) was obtained from Cayman Chemical (catalog no. 14965). Cycloheximide was purchased from Calbiochem (catalog no. 239764).

Stewart A.N., Tan S.Y., Clark D.J., Zhang H, & Wong G.W. (2019). N-Linked Glycosylation-Dependent and -Independent Mechanisms Regulating CTRP12 Cleavage, Secretion, and Stability. Biochemistry, 58(6), 727-741.

+ Open protocol

+ Expand

Gel-Based Protein Digestion Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Gel bands were diced into 1×1 mm squares, rinsed multiple times with 50 mM ammonium bicarbonate and reduced with 5 mM DTT, 50 mM ammonium bicarbonate at 55°C for 30 min. Residual solvent was removed and alkylation was performed using 10 mM propionamide in 50 mM ammonium bicarbonate for 30 min at room temperature. The gel pieces were rinsed 3 times with 50% acetonitrile, 50 mM ammonium bicarbonate and placed in a speed vac for 5 min. Digestion was performed with trypsin/LysC (Promega) in both a standard overnight digest (14 hr) at 37°C as well as in a limited digest format (1 hr at 50°C). Chymotrypsin (Promega) digests were performed overnight with the addition of 0.5 mM CaCl₂ to the 50 mM ammonium bicarbonate. Tubes were centrifuged and the solvent including peptides were collected and further peptide extraction was performed by the addition of 60% acetonitrile, 39.9% water, 0.1% formic acid and incubation for 10–15 min. The peptide pools were dried in a speed vac.

Komolov K.E., Du Y., Duc N.M., Betz R.M., Rodrigues J.P., Leib R.D., Patra D., Skiniotis G., Adams C.M., Dror R.O., Chung K.Y., Kobilka B.K, & Benovic J.L. (2017). Structural and Functional Analysis of a β2-Adrenergic Receptor Complex with GRK5. Cell, 169(3), 407-421.e16.

+ Open protocol

+ Expand

Proteomic Analysis of Modified Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

All reagents and solvents were purchased at the highest purity available. Cadaverine dihydrochloride, 2HP dihydrochloride, horseradish peroxidase (HRP), ethanol, 4-vinylpyridine, ammonium citrate, and citric acid were purchased from Sigma-Aldrich (St. Louis, MO, USA). Boric acid, sodium borate, and N-acetyl-3,7-dihydroxyphenoxazine (Amplex™ Red Reagent) were obtained from Thermo Fisher Scientific (Lenexa, KS, USA). Optima™ LC/MS-grade acetonitrile, water, and formic acid were purchased from Fisher Scientific (Thermo Fisher Scientific). Sequencing grade trypsin, chymotrypsin, and glycerol-free peptidyl-N-glycosidase F (PNGase F) were purchased from Promega (Madison, WI, USA) and New England BioLabs (Ipswich, MA, USA), respectively. Deionized water with a resistivity of 18 MΩ∙cm was purified using a Millipore Direct-Q3 Water Purification System (Billerica, MA, USA).

Meier A.A., Go E.P., Moon H.J., Desaire H, & Mure M. (2022). Mass Spectrometry-Based Disulfide Mapping of Lysyl Oxidase-like 2. International Journal of Molecular Sciences, 23(11), 5879.

+ Open protocol

+ Expand

Bromodomain 1 Binding Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

The bromodomain 1 (residue 41–168) of human BRD4 (refereed as BRD4 hereafter) and compound 1 were provided by Bristol Myers Squibb. L-Glutamine, L-methionine, catalase, hydrogen peroxide (H₂O₂), trifluoroacetic acid (TFA), formic acid (FA), phosphate buffer saline (PBS, 10 mM phosphate, 138 mM NaCl, 2.7 mM KCl), urea, dithiothreitol and iodoacetamide were from Sigma Aldrich (St. Louis, MO). Trypsin and chymoTrypsin were from Promega (Madison, WI).

Li K.S., Bergman E., Beno B.R., Huang R.Y., Deyanova E., Chen G, & Gross M.L. (2019). Hydrogen-Deuterium Exchange and Hydroxyl Radical Footprinting for Mapping Hydrophobic Interactions of Human Bromodomain with a Small Molecule Inhibitor. Journal of the American Society for Mass Spectrometry, 30(12), 2795-2804.

+ Open protocol

+ Expand

Ricin Digestion and LC-MS Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Firstly, 50 μL of purified ricin sample was transferred in a 200 μL PCR tube, which contains 10 μL of 1.0 M Tris-HCl (pH 8.0) and 10 μL acetonitrile. Subsequently, aliquot of 8 μL of 0.25 μg/μL chymotrypsin (Promega, Sequencing Grade) was added and then the mixture was incubated for 4 h at 40 °C. The digestion was terminated by adding 8 μL of 5% FA. The resulting sample was transferred to an autosampler vial for LC-MS analysis.

Liang L.H., Liu C.C., Chen B., Yan L., Yu H.L., Yang Y., Wu J.N., Li X.S, & Liu S.L. (2019). LC-HRMS Screening and Identification of Novel Peptide Markers of Ricin Based on Multiple Protease Digestion Strategies. Toxins, 11(7), 393.

+ Open protocol

+ Expand

HA-SS-np Stability Characterization

Check if the same lab product or an alternative is used in the 5 most similar protocols

HA-SS-np materials were produced at the Vaccine Production Program Laboratory (Gaithersburg, MD). HA-SS-np formulated in one of the five formulation buffers (containing sodium phosphate, sodium chloride, and sucrose, pH 7.2) was used for investigational purposes, and a HA-SS-np interim reference material (in 1x PBS) was used as a control. Reagents included LC-MS grade 0.1% formic acid in water (mobile phase A), 0.1% formic acid acetonitrile (mobile phase B), and ammonium bicarbonate that were purchased from JT Baker (Phillipsburg, NJ). RapiGest^™ surfactant and [Glu1]-Fibrinopeptide B lock mass standard were purchased from Waters (Milford, MA). Formic acid, urea, and Zeba^™ desalting columns (7K MWCO) were purchased from Pierce (Rockford, IL). Dithiothreitol (DTT) and iodoacetamide (IAM) were purchased from ThermoFisher Life Technologies (Grand Island, NY) and Sigma Aldrich (St. Louis, MO), respectively. Chymotrypsin and PNGase F glycosidase were purchased from Promega (Madison, WI); Lys-C was purchased from New England Biolabs (Ipswich, MA). Method details of the stability-indicating analyses (rCGE and SEC) are described in the online supporting information.

Schneck N.A., Ivleva V.B., Rosales-Zavala E., Wang X., Gollapudi D., Cooper J.W, & Lei Q.P. (2019). Using LC-MS to Identify Clipping in Self-Assembled Nanoparticles During Vaccine Development. Journal of the American Society for Mass Spectrometry, 30(12), 2576-2579.

+ Open protocol

+ Expand

Glycoprotein Analysis Using TCEP and Inhibitors

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tris(2-carboxyethyl)phosphine (TCEP) was obtained from Thermo Scientific (Rockford, IL). The mouse monoclonal anti-FLAG M2 antibody was purchased from Sigma-Aldrich (St. Louis, MO). Glucosamine, iodoacetamide, and tunicamycin were obtained from Sigma-Aldrich. Trypsin and chymoTrypsin were purchased from Promega (Madison, WI); hydrazine resin was from Bio-Rad (Hercules, CA), and peptide N-glycosidase F (PNGase F) and endoglycosidase H (Endo H) were from New England Biolabs (Ipswich, MA). The selective collagen prolyl 4-hydroxylase (CP4H) inhibitor [6-(5-ethoxycarbonyl-thiazol-2-yl)-nicotinic acid ethyl ester; also known as diethyl-pythiDC] was obtained from Aobious (Gloucester, MA).

Stewart A.N., Little H.C., Clark D.J., Zhang H, & Wong G.W. (2020). Protein Modifications Critical for Myonectin/Erythroferrone Secretion and Oligomer Assembly. Biochemistry, 59(29), 2684-2697.

+ Open protocol

+ Expand

Quantitative Proteomics of Tau and Amyloid

Check if the same lab product or an alternative is used in the 5 most similar protocols

Trypsin gold (mass spectrometry grade) and chymotrypsin were obtained from Promega (Madison, WI). Glass screw neck vials for liquid chromatography autosampler were from Waters (Milford, MA). Protease Inhibitor Cocktail (PIC), dithiothreitol (DTT), and iodoacetamide (IAA) were obtained from Sigma (St. Louis, MO). Acids and organic solvents were HPLC grade. Recombinant tau 441 and Beta-amyloid (1–42) standards unlabeled (quantified using amino acid analysis) and ¹⁵N tau 441 uniformly labeled and ¹⁵N Beta-amyloid (1–42) uniformly labeled were purchased from rPeptide (Bogart, GA).

Pottiez G., Yang L., Stewart T., Song N., Aro P., Galasko D., Quinn J., Peskind E., Shi M, & Zhang J. (2017). A mass spectrometry-based method to quantify in parallel Tau and amyloid β 1–42 in CSF for the diagnostic of Alzheimer’s Disease. Journal of proteome research, 16(3), 1228-1238.

+ Open protocol

+ Expand

SARS-CoV-2 Spike Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Dithiothreitol (DTT) and iodoacetamide (IAA) were purchased from Sigma Aldrich (St. Louis, MO). Sequencing-grade modified trypsin and chymotrypsin were purchased from Promega (Madison, WI). All other reagents were purchased from Sigma Aldrich unless indicated otherwise. Data analysis was performed using Byonic 3.5 software and manually using Xcalibur 4.2. The SARS-CoV-2 spike protein culture supernatant subunit S1 (Cat. No. 230-20407) and subunit S2 (Cat. No. 230-20408), and purified subunit S2 (Cat. No. 230-30163) were purchased from RayBiotech (Atlanta, GA).

Shajahan A., Supekar N.T., Gleinich A.S, & Azadi P. (2020). Deducing the N- and O-glycosylation profile of the spike protein of novel coronavirus SARS-CoV-2. Glycobiology, 30(12), 981-988.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!