| Description | Carboxypeptidase B, also known as peptidyl - L - lysine (L - arginine) hydrolase and arginase, is a metalloprotease that can specifically hydrolyze basic amino acids (lysine, arginine, histidine) at the C - terminus of proteins.This product is a recombinant carboxypeptidase B lyophilized powder Carboxypeptidase B, also known as peptidyl - L - lysine (L - arginine) hydrolase and arginase, is a metalloprotease that can specifically hydrolyze basic amino acids (lysine, arginine, histidine) at the C - terminus of proteins.This product is a recombinant carboxypeptidase B lyophilized powder obtained by isolation and purification from recombinant Pichia pastoris, followed by freeze - drying. It is a biochemical - grade genetic engineering enzyme, free from exogenous virus contamination, and does not contain enzyme inhibitors such as DFP, PMSF, and TLCK. It has the same activity and specificity as natural carboxypeptidase B.Product AdvantagesAnimal - free origin: No animal viruses, no pathogenic substances, no exogenous factor contamination, and high safety.High purity and high specific activity.Production scale is over 1000L.Compliance: Production equipment and production environment meet relevant regulatory requirements and conform to GMP guidelines.Complete quality documents: Relevant regulatory support documents can be provided according to customer needs.Product CharacteristicsAppearance: White or off - white powderEnzyme specific activity: ≥150 U/mg proteinMolecular weight: 36kD ± 3.6kDPurity: ≥90% (SDS - PAGE)Protein content: ≥30%Optimal pH: 7.0 - 9.0Solubility: Soluble in buffer solutionRecommended Usage MethodsPreparation of proteinDissolve the protein in a 25mM Tris - HCl solution with pH 7.6.Preparation of enzyme solutionWeigh the required enzyme powder and dissolve it in 25mM Tris - HCl (pH 7.6) to make the enzyme solution concentration 1 - 10mg/ml.ReactionRecommended enzyme amount: Target protein (mass ratio) = 1:50 - 1:1000, and the optimal pH is 7.0 - 9.0.Product ApplicationsProduction of various proteins or polypeptides, such as insulin and its analogs, beauty peptides, antimicrobial peptides, food flavor peptides, etc.Quality detection of antibodies.Determination of amino acids at the C - terminus of proteins.Storage stabilityThe lyophilized powder can be stably stored for at least 24 months at -15°C or below. After being dissolved in 25 mM Tris-HCl (pH 7.6) and stored at -15°C or below, there is no loss of activity even after 10 cycles of repeated freezing and thawing. It is recommended that the dissolved product be stored at 2–8°C for no more than 7 days, and at -15°C or below for no more than 30 days.Precautions and DisclaimerIts activity is competitively inhibited by arginine and lysine; metal ion chelators (such as EDTA) also have an inhibitory effect on the enzyme activity.This product is hygroscopic and should be equilibrated at room temperature before being taken and used.This product is for research use only and shall not be used for the diagnosis or treatment of animals or humans... Read More | Inquire | Malic Dehydrogenase is a ubiquitous enzyme, which exists in two isoforms in eukaryotic cells.Malic dehydrogenase exists as a dimer with each subunit containing an NAD-binding domain and a substrate-binding carboxy-terminal domain required for activity. Malic dehydrogenase is a cytoplasmic isozyme Malic Dehydrogenase is a ubiquitous enzyme, which exists in two isoforms in eukaryotic cells.Malic dehydrogenase exists as a dimer with each subunit containing an NAD-binding domain and a substrate-binding carboxy-terminal domain required for activity. Malic dehydrogenase is a cytoplasmic isozyme and an important catalyst in the tricarboxylic acid cycle.ReagentsA. 0.1 M Tris-HCl buffer (pH7.8)B. 0.01 M Phosphate buffer (KH2PO4-NaOH, pH 7.0)C. Triton X-100 solution (50 mg/ml)D. 0.01 M Phosphate buffer containing 0.1% Triton X-100 (KH2PO4-NaOH, pH 7.0)Dilute 20 ml of Triton X-100 solution (C) with approx. 800 ml of 0.01M Phosphate buffer (B). Fill up to 1,000 ml with 0.01M Phosphate buffer (B).E. NADH soluton Weigh 9 mg of NADH and dissolve in 0.1M Tris-HCl bufer (A). Fill up to 50 ml with 0.1M Tris-HCl Buffer (A). (Can be used for 5 days if kept refrigerated)F. Substrate solutionWeigh 11 mg of oxaloacetic acid and dissolve in 0.1M Tris-HCl buffer (A). Fill up to 50 ml with 0.1M Tris-HCl buffer (A) (Make a fresh solution for each use.)G. Enzyme solutionWeigh out Malate Dehydrogenase and dissolve in chilled 0.01M Phosphate Bufer containing 0.1% Triton X-100 (D). Enzyme solution should be prepared so that the value of AOD/minute becomes in the range of 0.025 ± 0.010.ProcedurePipette 2.0 ml of NADH solution (E) and 0.90 ml of Substrate solution (F) respectively into a quartz cell (d=10 mm) and keep at 25 + 0.5'℃ for 5 minutes. Then, pipete 0.10 ml of Enzyme solution (G) into the quartz cell and mix well immediately. Keep the reaction mixture at 25 ±0.5'C.Exaclly at 2 minutes and 5 minutes after the addition of Enzyme solution (G), measure the absorbances of the reaction mixture at 340 nm(A2 and A5).As a blank, pipette 0.01M Phosphate buffer (D) into another quartz cel (d=10 mm) instead of the Enzyme solution (G) and follow the same procedure described above (Ab2 and Ab5).CalculationMalate dehydrogenase activity (u/mg)=[(A2-A5)-(Ab2-Ab5)]/3*(1/6.22)*(n/0.1) ApplicationThis enzyme is used for the enzymatic determination of L-malate and gluamate oxalo-acetate transaminase(GOT)in clinical diagnosis... Read More | Purity>97% SDS-PAGE and HPLC analyses. FunctionLA-PF4 stimulates DNA synthesis, mitosis, glycolysis, intracellular cAMP accumulation, prostaglandin E2 secretion, and synthesis of hyaluronic acid and sulfated glycosaminoglycan. It also stimulates the formation and secretion of plasminogen Purity>97% SDS-PAGE and HPLC analyses. FunctionLA-PF4 stimulates DNA synthesis, mitosis, glycolysis, intracellular cAMP accumulation, prostaglandin E2 secretion, and synthesis of hyaluronic acid and sulfated glycosaminoglycan. It also stimulates the formation and secretion of plasminogen activator by human synovial cells. NAP-2 is a ligand for CXCR1 and CXCR2, and NAP-2, NAP-2(73), NAP-2(74), NAP-2(1-66), and most potent NAP-2(1-63) are chemoattractants and activators for neutrophils. TC-1 and TC-2 are antibacterial proteins, in vitro released from activated platelet alpha-granules. CTAP-III(1-81) is more potent than CTAP-III desensitize chemokine-induced neutrophil activation.Post-translationalProteolytic removal of residues 1-9 produces the active peptide connective tissue-activating peptide III (CTAP-III) (low-affinity platelet factor IV (LA-PF4)). Proteolytic removal of residues 1-13 produces the active peptide beta-thromboglobulin, which is released from platelets along with platelet factor 4 and platelet-derived growth factor. NAP-2(1-66) is produced by proteolytical processing, probably after secretion by leukocytes other than neutrophils. NAP-2(73) and NAP-2(74) seem not be produced by proteolytical processing of secreted precursors but are released in an active form from platelets... Read More | Inquire |