| Description | Reconstitution: After centrifugation, we suggest addition of 50 mM Na Acetate, pH 5.5, with 150 mM NaCl to the original volume, followed by gentle mixing to ensure adequate homogenization.This enzyme degrades elastin, collagen, and proteoglycan and has been implicated in the development of pulmonaryReconstitution: After centrifugation, we suggest addition of 50 mM Na Acetate, pH 5.5, with 150 mM NaCl to the original volume, followed by gentle mixing to ensure adequate homogenization.This enzyme degrades elastin, collagen, and proteoglycan and has been implicated in the development of pulmonary emphesema and rheumatoid arthritis.Activity: 20-22 units per mg protein. One unit is defined as the amount of enzyme that hydrolyzes one umole of MeO-Suc-Ala-Ala-Pro-Val-pNA per minute at 25℃, pH 7.5. Product Citations:Mittendorf, Elizabeth A., Gheath Alatrash, Na Qiao, Yun Wu, Pariya Sukhumalchandra, Lisa S. St John, Anne V. Philips et al. "Breast cancer cell uptake of the inflammatory mediator neutrophil elastase triggers an anticancer adaptive immune response." Cancer research 72, no. 13 (2012): 3153-3162van den Berg, Carmen W., et al. "Mechanism of Neutrophil Dysfunction: Neutrophil Serine Proteases Cleave and Inactivate the C5a Receptor." The Journal of Immunology 192.4 (2014): 1787-1795.Edwards, J. Vincent, Nicolette Prevost, Kandan Sethumadhavan, Abul Ullah, and Brian Condon. "Peptide conjugated cellulose nanocrystals with sensitive human neutrophil elastase sensor activity." Cellulose (2013): 1-13.Valdez C, Wong YC, Schwake M, Bu G, Wszolek ZK, Krainc D.Progranulin-mediated deficiency of cathepsin D results in FTD and NCL-like phenotypes in neurons derived from FTD patients.Hum Mol Gen. 2017. 2017 Sep 25. doi:10.1093/hmg/ddx364Kerros C, et al.Neuropilin-1 mediates neutrophil elastase uptake and cross-presentation in breast cancer cells.J Biol Chem. 2017 Jun 16;292(24):10295-10305. doi: 10.1074/jbc.M116.773051. Epub 2017 May 3.Martin KR, Kantari-Mimoun C, Yin M, Pederzoli-Ribeil M, et al.Proteinase 3 Is a Phosphatidylserine-binding Protein That Affects the Production and Function of Microvesicles.J Biol Chem. 2016 May 13;291(20):10476-89. doi: 10.1074/jbc.M115.698639. Epub 2016 Mar 9.Edwards JV, Fontenot KR, Prevost NT, Pircher N, Liebner F, Condon BD.Preparation, Characterization and Activity of a Peptide-Cellulosic Aerogel Protease Sensor from Cotton.Sensors (Basel). 2016 Oct 26;16(11). pii: E1789.Chawla A, Alatrash G, Philips AV, Qiao N, et al.Neutrophil elastase enhances antigen presentation by upregulating human leukocyte antigen class I expression on tumor cells.Cancer Immunol Immunother. 2016 Jun;65(6):741-51. doi: 10.1007/s00262-016-1841-6. Epub 2016 Apr 29.Sinden NJ, et al.α-1-Antitrypsin variants and the proteinase/antiproteinase imbalance in chronic obstructive pulmonary disease.Am J Physiol Lung Cell Mol Physiol. 2015 Jan 15;308(2):L179-90.Ksiazek M, et al.Miropin, a Novel Bacterial Serpin from the Periodontopathogen Tannerella forsythia, Inhibits a Broad Range of Proteases by Using Different Peptide Bonds within the Reactive Center Loop.J Biol Chem. 2015 Jan 2;290(1):658-70. doi: 10.1074/jbc.M114.601716. Epub 2014 Nov 11.Ramadass M, Ghebrehiwet B, Kew RR.Enhanced recognition of plasma proteins in a non-native state by complement C3b. A possible clearance mechanism for damaged proteins in blood.Mol Immunol. 2015 Mar;64(1):55-62. doi: 10.1016/j.molimm.2014.10.022. Epub 2014 Nov 15.Thomas MP, et al.Leukocyte Protease Binding to Nucleic Acids Promotes Nuclear Localization and Cleavage of Nucleic Acid Binding Proteins.J Immunol. 2014 Jun 1;192(11):5390-7. doi: 10.4049/jimmunol.1303296. Epub 2014 Apr 25.Ungurs MJ, Sinden NJ, Stockley RA.Progranulin is a substrate for neutrophil-elastase and proteinase-3 in the airway and its concentration correlates with mediators of airway inflammation in COPDAm J Physiol Lung Cell Mol Physiol. 2014 Jan 1;306(1):L80-7. doi: 10.1152/ajplung.00221.2013. Epub 2013 Nov 1.Sinden NJ, Koura F, Stockley RA.The significance of the F variant of alpha-1-antitrypsin and unique case report of a PiFF homozygote.BMC Pulm Med. 2014 Aug 7;14:132. doi: 10.1186/1471-2466-14-132.Kenninston JA, et al.Inhibition of Plasma Kallikrein by a Highly Specific Active Site Blocking Antibody.J Biol Chem. 2014 Aug 22;289(34):23596-608. doi: 10.1074/jbc.M114.569061. Epub 2014 Jun 26.Tejera P, et al.Functional Characterization of Polymorphisms in the Peptidase Inhibitor 3 (Elafin) Gene and Validation of Their Contribution to Risk of Acute Respiratory Distress Syndrome.Am J Respir Cell Mol Biol. 2014 Aug;51(2):262-72. doi: 10.1165/rcmb.2013-0238OC.Zheng K, et al.Influence of glycosylation pattern on the molecular properties of monoclonal antibodies.MAbs. 2014 May-Jun;6(3):649-58. doi: 10.4161/mabs.28588. Epub 2014 Mar 24... Read More | Inquire | Inquire | Product DescriptionEndo F1 cleaves Asparagine-linked high mannose and some hybrid oligosaccharides. Core fucosylation reduces the activity by 50 fold. Endoglycosidase F1 will hydrolyze sulfate containing high-mannose chains. It cleaves between the two N-acetylglucosamine residues in the Product DescriptionEndo F1 cleaves Asparagine-linked high mannose and some hybrid oligosaccharides. Core fucosylation reduces the activity by 50 fold. Endoglycosidase F1 will hydrolyze sulfate containing high-mannose chains. It cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact.Molecular weight 32,000 daltonsContents60 µl aliquot of enzyme (1 U) in 20 mM Tris-HCl, pH 7.5Included with 20 µL and 60 µL pack sizes:5x Reaction Buffer – 250 mM sodium phosphate, pH 5.5Specific ActivityDefined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 micromole of denatured Ribonuclease B (RNase B) in 1 minute at 37°C, pH 5.5. Cleavage is monitored by SDS-PAGE (cleaved RNase B migrates faster).FormulationThe enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl, pH 7.5StabilitySeveral days exposure to ambient temperatures will not reduce activity. Stable at least 12 months when stored properly. SpecificityEndo F1 cleaves Asparagine-linked high mannose or hybrid oligosaccharides. It cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact. Quality & PurityEndo F1 is tested for contaminating protease as follows: 10 µg of denatured BSA is incubated at 37°C for 24 hours with 2 µl of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation. The production host strain has been extensively tested and does not produce any detectable glycosidases.Directions for use1. Add up to 200 µg of glycoprotein to an Eppendorf tube. Adjust to 38 µl final volume with de-ionized water.2. Add 10 µl 5x Reaction Buffer 5.53. Add 2.0 µl of Endo F1 to the reaction. Incubate 1 hour or more at 37°C.Monitor cleavage by SDS-PAGE... Read More | Inquire |