| Description | If the serum will not be used up within a short period of time, dispense after thawing and store aliquot under appropriate conditions for long-term storage. The volume of serum will increase by 10% after freezing, so when dispensing serum into tubes, leave some space in the tube to avoid tube If the serum will not be used up within a short period of time, dispense after thawing and store aliquot under appropriate conditions for long-term storage. The volume of serum will increase by 10% after freezing, so when dispensing serum into tubes, leave some space in the tube to avoid tube cracking during the freezing.After the serum is thawed completely, the serum can be heated at 56℃ for 30 minutes to inactivate the complement in serum. Unless necessary, heat-inactivation of the serum is not recommended for general use because heat treatment will result in precipitation of serum and decreasing of serum quality. Complement is involved in reactions such as cytotoxicity, contraction of smooth muscle cells, release of histamine from mast cells and platelets, enhancement of phagocytosis, and promotion of chemical chemotaxis and activation of lymphocytes and macrophages.The serum should be thawed slowly and gradually. Thaw the serum stored at -15~-40℃ in 4℃ refrigerator for approximately one day, and make aliquot after it is thawed thoroughly. During the thawing process, shake gently every 2 hours (avoid the formation of air bubbles) to homogenize the component and reduce the occurrence of precipitation. Do not thaw the serum stored at -20℃ or lower temperatures directly in a 37℃ water bath to avoid protein agglomeration and precipitation in serum due to large temperature changes.The floc-like precipitates in serum are mainly denatured fibrin and lipoproteins in serum after long-term storage at 4℃. These flocs do not affect the quality of the serum and can be left untreated. If necessary, the flocs can be removed by centrifugation at 400×g for 5 minutes. However, it is not advisable to remove by filtration as the flocs may block the filter membrane. The precipitates in the serum after heat inactivation will increase significantly. Some of these deposits look like "black dots" under the microscope and are often misinterpreted as microbial contamination because the Brownian motion of these "dots" is magnified under the microscope and appear to be swimming. Usually, these small black spots do not affect cell growth, but if microbial contamination of the serum is suspected, it should be immediately replaced with new serum. Users can dilute the serum with culture medium to 10% concentration and incubate for 1-3 days, and observe whether the small black dots increase sharply, or spread an appropriate amount of serum on LB plate to determine whether there is microbial contamination. Do not leave the serum at 37℃ for too long, as the serum will gradually become cloudy and lose some active ingredients, thus affecting the quality of the serum.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation... 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 |