| Description | This product is a premixed system composed of GoldStar Best DNA Polymerase, Mg2+, dNTPs, PCR stabilizers and enhancers, with a concentration of 2 ×. It has the advantages of simple and fast operation, high sensitivity, strong specificity, and good stability, which can minimize human error and This product is a premixed system composed of GoldStar Best DNA Polymerase, Mg2+, dNTPs, PCR stabilizers and enhancers, with a concentration of 2 ×. It has the advantages of simple and fast operation, high sensitivity, strong specificity, and good stability, which can minimize human error and pollution. This polymerase has 5 '-3' DNA polymerase activity, 5 '-3' exonuclease activity, and 3 '-5' exonuclease activity. Under normal PCR conditions, compared with GoldStar DNA polymerase, it has excellent performance of high amplification efficiency and low mismatch rate. The chemically modified enzyme does not exhibit polymerase activity at room temperature, effectively avoiding non-specific amplification caused by non-specific binding of primers and templates or primer dimers at room temperature. Enzyme activation requires incubation at 95 ℃ for 10 minutes, which can be integrated into existing PCR thermal cycling programs. The optimized buffer system maximizes the effectiveness of the enzyme, achieving high fidelity, specificity, amplification efficiency, and sensitivity for the target fragment. This product does not contain dyes. After the PCR program is completed, an appropriate amount of sample buffer can be added as needed for electrophoresis operation. Most PCR products obtained from amplification have an "A" base attached to the 3 'end, making them suitable for direct use in T/A cloning. Suitable for routine PCR reactions and gene cloning experiments that require high fidelity.Note: 2 x GoldStar Best MasterMix contains GoldStar Best DNAPolymerase, 3.4 mM MgCl2, and 400 µ M each dNTP.G665561Component1 mL5 mL25 mLStorageG665561A2×GoldStar Best MasterMix1 mL5×1 mL5×5 mL-20℃.Avoid freeze/thaw cycle. G665561BddH2O 1 mL5×1 mL5×5 mL-20℃.Avoid freeze/thaw cycle. Notice: 2×GoldStar Best MasterMix contains GoldStar Best DNA Polymerase, 3.4 mM MgCl2 and 400 µM each dNTP.Quality control:After testing, there was no exogenous nuclease activity; PCR method for detecting residual DNA without host; Can effectively amplify single copy genes from multiple genomes; Storage at 2-8 ℃ for three months showed no significant change in activity.Usage:The following is an example of a PCR reaction system and reaction conditions for amplifying a 1 kb fragment using human genomic DNA as a template. In practical operation, corresponding improvements and optimizations should be made based on the template, primer structure, and target fragment size.1. PCR reaction systemReagent50 µlReaction systemFinal concentration2×GoldStar Best MasterMix（Dye）25 µl1×Forward Primer，10 µM2 µl0.4 µMReverse Primer，10 µM2 µl0.4 µMTemplate DNA<0.5 µg<0.5 µg/50 µlddH2Oup to 50 µl/Attention: The primer concentration should be between 0.1 and 1.0 as the final concentration µ M serves as a reference for setting the range. In the case of low amplification efficiency, the concentration of primers can be increased; When non-specific reactions occur, the primer concentration can be reduced to optimize the reaction system.2. PCR reaction conditionsStepTemperatureTime/Pre denaturation95℃10 min/Denaturation94℃30 s30-40 cyclesAnneal55-65℃30 s30-40 cyclesExtend72℃60 s30-40 cyclesFinally extended72℃5 min/Attention:1) In general experiments, if the annealing temperature is 5 ℃ lower than the melting temperature Tm of the amplification primer, and the ideal amplification efficiency cannot be achieved, the annealing temperature should be appropriately reduced; When non-specific reactions occur, increase the annealing temperature to optimize the reaction conditions.2) The extension time should be set according to the size of the amplified fragment. The amplification efficiency of GoldStar Best DNA Polymerase contained in this product is 1-2 kb/min.3) The number of cycles can be set based on the downstream application of the amplification product. If the number of cycles is too small, the amplification amount is insufficient; If there are too many cycles, the probability of mismatches will increase, and non-specific backgrounds will be severe. So, while ensuring product yield, the number of cycles should be minimized as much as possible.4) This product must achieve enzyme activation under pre denaturation conditions of 95 ℃ and 10 minutes... Read More | Inquire | Product DescriptionEndo F2 cleaves N-linked (asparagine-linked) biantennary oligosaccharides from glycoproteins. It also will cleave high mannose glycans but at a 40x reduced rate. It cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, Product DescriptionEndo F2 cleaves N-linked (asparagine-linked) biantennary oligosaccharides from glycoproteins. It also will cleave high mannose glycans but at a 40x reduced rate. 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.Endoglycosidase F2 is less sensitive to protein conformation than PNGase F and is therefore more suitable for deglycosylation of native proteins. However, for optimal results, denaturation of the glycoprotein is recommended.Contents60 µl aliquot of enzyme (0.3 U) in 10 mM sodium acetate 25mM NaCl, pH 4.5Included with 20 µL and 60 µL pack sizes:5x Reaction Buffer – 250 mM sodium acetate, pH 4.5Molecular weight 32,000 daltonsSpecific Activity Defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 micromole of denatured porcine fibrinogen in 1 minute at 37°C, pH 5.5. Cleavage is monitored by SDS-PAGE (cleaved fibrinogen migrates faster).Formulation The enzyme is provided as a sterile-filtered solution in 10 mM sodium acetate, 25mM NaCl, pH 4.5Specificity Endo F2 cleaves Asparagine-linked biantennary and high mannose glycans (at a 40X reduced rate). 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. Endoglycosidase F2 is less sensitive to protein conformation than PNGase F and is therefore more suitable for deglycosylation of native proteins. However for optimal results, denaturation of the glycoprotein is recommended.Quality & Purity Endo F2 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.Stability Several days exposure to ambient temperatures will not reduce activity. Stable at least 12 months when stored properly.Directions for use 1. 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 4.5 3. Add 2.0 µl of Endo F2 to the reaction. Incubate 1 hour at 37°C. Monitor cleavage by SDS-PAGEThe production host strain has been extensively tested and does not produce any detectable glycosidases... Read More | Inquire | Parathyroid Hormone (1-34), human, biotinylated is a probe for the parathyroid hormone receptor, can be used for analyzing the interaction between parathyroid hormone and parathyroid hormone receptors in living cells and for purifying hormone-receptor comAppearance:SolidBiological Activity:Parathyroid Hormone (1-34), human, biotinylated is a probe for the parathyroid hormone receptor, can be used for analyzing the interaction between parathyroid hormone and parathyroid hormone receptors in living cells and for purifying hormone-receptor comAppearance:SolidBiological Activity:Parathyroid Hormone (1-34), human, biotinylated is a probe for the parathyroid hormone receptor, can be used for analyzing the interaction between parathyroid hormone and parathyroid hormone receptors in living cells and for purifying hormone-receptor com... Read More |