Pepstatin a

Manufactured by MP Biomedicals

Pepstatin A is a peptide inhibitor that acts on aspartic proteases, such as pepsin, renin, and some cathepsins. It is commonly used in research applications to prevent the action of these enzymes.

Automatically generated - may contain errors

Lab products found in correlation

Pierce ecl, by Thermo Fisher Scientific (3 mentions) Vitamin k, by Merck Group (2 mentions) Enzchek protease activity assay, by Thermo Fisher Scientific (2 mentions) Dmso solubilized, by MP Biomedicals (2 mentions) Hemin, by Merck Group (2 mentions) Amicon ultra 15 filter, by Merck Group (2 mentions) Gm6001, by Merck Group (2 mentions) Chloroquine, by Merck Group (1 mentions) N trans epoxysuccinyl l leucine 4 guanidinobutylamide e 64, by Merck Group (1 mentions) Clindamycin, by Merck Group (1 mentions) G7 reaction buffer, by New England Biolabs (1 mentions) Np 40, by New England Biolabs (1 mentions) Glycoprotein denaturing buffer, by New England Biolabs (1 mentions) G5 reaction buffer, by New England Biolabs (1 mentions) Phenyl methyl sulfonyl fluoride (pmsf), by New England Biolabs (1 mentions) Endo h, by New England Biolabs (1 mentions) Pngase f, by New England Biolabs (1 mentions) Phenyl methyl sulfonyl fluoride (pmsf), by MP Biomedicals (1 mentions) Ab13824, by Abcam (1 mentions) Pierce bca method, by Thermo Fisher Scientific (1 mentions)

7 protocols using pepstatin a

TAMRA-fluorophosphonate Probe Activity

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

The TAMRA-fluorophosphonate activity-based probe was obtained from ThermoFisher. N-(trans-epoxysuccinyl)-l-leucine-4-guanidinobutylamide (E-64), AA74-1, clindamycin, chloroquine, and isopentenyl pyrophosphate trilithium salt were purchased from Sigma. Pepstatin A was purchased from MP Biomedicals.

Elahi R., Dapper C, & Klemba M. (2019). Internalization of Erythrocyte Acylpeptide Hydrolase Is Required for Asexual Replication of Plasmodium falciparum. mSphere, 4(3), e00077-19.

+ Open protocol

+ Expand

Assessing Bacteroides vulgatus Protease Inhibition

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

Bacteroides vulgatus (ATCC 8482) were grown anaerobically in Brain-heart-infusion (BHI, BD) broth supplemented with 5 μg/ml hemin (Sigma) and 0.5 μg/ml vitamin K (Sigma). Overnight supernatant was collected by pelleting cells at 8000 x g. Supernatant was then 8-fold concentrated at 3,300 x g for 15 minutes using 10 kDa Amicon Ultra-15 filters (Millipore). Concentrated supernatant protease activity was tested using the EnzChek protease activity assay (Invitrogen) after incubation for 24 hours at 37 °C measuring fluorescence at 485 nm for excitation and 530 nm for emission. Protease inhibitors were administered at 10% total volume and inhibition was calculated by comparison to vehicle control wells. Protease inhibitors tested included water-solubilized 4(2-Aminoethyl)benzenesulfonyl Fluoride (AEBSF, MP Biomedicals), water-solubilized E-64 (Sigma), DMSO-solubilized GM6001 (EMD Millipore), and DMSO-solubilized Pepstatin A (MP Biomedicals). After analysis of a preliminary dilution series, max inhibition was found for each protease inhibitor at the highest concentration allowed by the solubility of each compound, and these concentrations were used for subsequent studies.

Mills R.H., Dulai P.S., Vázquez-Baeza Y., Sauceda C., Daniel N., Gerner R.R., Batachari L.E., Ochoa M.M., Zhu Q., Weldon K., Humphrey G., Carrillo-Terrazas M., Goldasich L.D., Bryant M., Raffatellu M., Quinn R.A., Gewirtz A.T., Chassaing B., Chu H., Sandborn W.J., Dorrestein P.C., Knight R, & Gonzalez D.J. (2022). Multi-omics analyses of the Ulcerative Colitis gut microbiome link Bacteroides vulgatus proteases with disease severity. Nature microbiology, 7(2), 262-276.

+ Open protocol

+ Expand

Glycosylation analysis of pulse-chase fractions

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

Immunoprecipitates of pulse-chase fractions were boiled in Glycoprotein Denaturing Buffer (NEB, Ipswich, MA) for 10 minutes. For wild type cells, immunoprecipitates of the intracellular fraction at time 0 and 0.5 hours (IC 0 and IC 0.5) and extracellular fraction at time 4.0 hours (EC 4.0) were analyzed. For ATD cells, immunoprecipitates of the intracellular fraction at time 0 and 4.0 hours (IC 0 and IC 4.0) and extracellular fraction at time 4.0 hours (EC 4.0) were analyzed. Samples were centrifuged at 13200 rpm for 10 minutes. For endo H, supernatants were added to a reaction containing 1X G5 Reaction Buffer (NEB, Ipswich, MA), 1 mM PMSF (MP Biomedicals, Solon, OH), 10 uM pepstatin A (MP Biomedicals, Solon, OH), and 20 U/uL endo H (NEB, Ipswich, MA) and incubated at 37°C for 1 hour. For PNGase F, supernatants were added to a reaction containing 1X G7 Reaction Buffer (NEB, Ipswich, MA), 1% NP-40 (NEB, Ipswich, MA), 1 mM PMSF, 10 uM pepstatin A, and 10 U/uL PNGase F (NEB, Ipswich, MA) and incubated at 37°C for 1 hour. Samples were analyzed by analyzed by SDS-PAGE/fluorography.

Tafaleng E.N., Chakraborty S., Han B., Hale P., Wu W., Soto-Gutierrez A., Feghali-Bostwick C.A., Wilson A.A., Kotton D.N., Nagaya M., Strom S.C., Chowdhury J.R., Stolz D.B., Perlmutter D.H, & Fox I.J. (2015). Induced pluripotent stem cells model personalized variations in liver disease due to α1-antitrypsin deficiency. Hepatology (Baltimore, Md.), 62(1), 147-157.

+ Open protocol

+ Expand

Fractionation and Western Blot Analysis of Aurora-A in Ovarian Tumors

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

Samples of normal fimbriae and ovarian tumors were obtained from fresh surgical specimens, snap frozen in liquid nitrogen and stored in − 80 degrees Celsius. Frozen tissue was homogenized on ice and processed into nuclear and cytosolic fractions as previously described [17 (link)]. Protease inhibitors (aprotinin (Sigma, St. Louis, MO), leupeptin, pepstatin A, chymostatin, and AEBSF (MP Biomedicals, Solon, OH)) and 1 μM DTT were added to all lysis buffers. Protein concentration was determined by the Pierce BCA method (Thermo Fisher Scientific, Waltham, MA). Nuclear and cytosolic fractions (20 μg total protein per lane) were electrophoresed on 10% polyacrylamide gels and electrotransferred to PVDF membranes (Millipore, Billerica, MA). Blocking was performed with 4% dried milk in TBST. Membranes were incubated with anti-human Aurora-A antibodies (Abcam, ab13824, 1:500), anti-human phospho- Thr²⁸⁸-Aurora-A (Abcam, ab58494, 1:500) or anti-human β-actin (Sigma, A2228, 1:5000) antibodies, followed by incubation with goat anti-mouse IgG-HRP (Santa Cruz Biotechnologies, Santa Cruz, CA) secondary antibody. Blots were developed with Pierce ECL (Thermo Fisher Scientific, Waltham, MA) and exposed to X-ray film.

Alkhateeb K.J., Crane J.E., Sak M., Jorgensen C.J., O’Donnell J.P., Zumbar C.T., Wozniak J.A., Salazar C.R., Parwani A.V, & Lehman N.L. (2021). Aurora-A kinase is differentially expressed in the nucleus and cytoplasm in normal Müllerian epithelium and benign, borderline and malignant serous ovarian neoplasms. Diagnostic Pathology, 16, 98.

+ Open protocol

+ Expand

Cellular Pathway Modulation Protocol

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

Chloroquine
and pepstatin A were purchased
from MP Biomedicals. Doxorubicin was purchased from Sigma-Aldrich.
Rapamycin and MG-132 were purchased from Calbiochem/EMD Biosciences.
Cytosine arabinoside was from Tocris Bioscience. Torin 1 was from
Cayman Chemical. The target sequences for shRNAs are as follows: human
CIMPR shRNA, CTACCTGTATGAGATCCAA; human VPS26A shRNA, CTCTATTAAGATGGAAGTG;
luciferase shRNA, GCACTCTGATTGACAAATACGATTT. Cathepsin D and firefly
luciferase–EWS-Fli-1 fusion cDNAs were cloned into pCDF1 lentiviral
vector (System Biosciences).

Elzi D.J., Song M., Hakala K., Weintraub S.T, & Shiio Y. (2014). Proteomic Analysis of the EWS-Fli-1 Interactome Reveals the Role of the Lysosome in EWS-Fli-1 Turnover. Journal of Proteome Research, 13(8), 3783-3791.

+ Open protocol

+ Expand

Assessing Bacteroides vulgatus Protease Inhibition

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

+ Open protocol

+ Expand

Isolation of Bovine Brain Proteins

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

Fresh bovine brains were obtained from J.W. Treuth & Sons (Baltimore, MD, USA). All chemicals were purchased from Sigma, unless designated otherwise. Leupeptin hemisulfate and pepstatin A were from MP Biomedicals. Tubes and rotors were from Beckman-Coulter. Dounce homogenizers were from Wheaton. Detergents were purchased from Avanti or Anatrace. Lipids were purchased from Avanti. NuPage Novex 4–12% Bis-Tris protein gels (NP0322BOX), PageRuler Plus prestained protein ladder (10–250 kDa) (26619), Pierce BCA protein assay kit (23225), and Protein G Dynabeads (10003D) were purchased from Thermo Scientific.

Weston M.R, & Mindell J.A. (2018). Characterizing chloride-dependent acidification in brain Clathrin-coated vesicles. Biochemistry and cell biology = Biochimie et biologie cellulaire, 97(3), 315-324.

+ 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!