DNA Polymerase I, Large (Klenow) Fragment

Name: DNA Polymerase I, Large (Klenow) Fragment
Brand: BetaLifeSci
SKU: ENY-104
Availability: InStock

Collections: Featured enzyme molecules, Tool enzymes & other reagents

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Product Overview

Description	DNA Polymerase I, Large (Klenow) Fragment (about 68 kD) is a proteolytic product of E. coli DNA Polymerase I which retains polymerization and 3'→5' exonuclease activity, but has lost 5'→3' exonuclease activity. Klenow retains the polymerization fidelity of the holoenzyme without degrading 5' termini. It is applicable to DNA sequencing by the Sanger dideoxy method, fill-in of 5' overhangs to form blunt ends, removal of 3' overhangs to form blunt ends, second strand cDNA synthesis and second strand synthesis in mutagenesis protocols. Theoretical protein molecular weight is 68000 daltons. Error Rate: ~ 18x10-6 bases
Source	An E.coli strain that contains the E. coli polA gene that has had its 5'→3' exonuclease domain removed.
Applications	Blunting, DNA Sequencing , Polymerases for DNA Manipulation, RT-qPCR, RT-PCR and cDNA Synthesis, RT-PCR & cDNA Synthesis, PCR
Size	200 U / 1000 U / 5000 U
Concentration	5,000 U/ml
Components	DNA Polymerase I, Large (Klenow) Fragment (5,000 U/ml); 10X ABuffer B
Unit Definition	One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 37°C.
Usage	For Research Use Only
Storage	Storage condition: 25 mM Tris-HCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.4 @25°C. Store the DNA Polymerase I, Large (Klenow) Fragment at -20°C. Please avoid repeated freeze-thaw cycles.

FAQs

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Proteins are sensitive to heat, and freeze-drying can preserve the activity of the majority of proteins. It improves protein stability, extends storage time, and reduces shipping costs. However, freeze-drying can also lead to the loss of the active portion of the protein and cause aggregation and denaturation issues. Nonetheless, these adverse effects can be minimized by incorporating protective agents such as stabilizers, additives, and excipients, and by carefully controlling various lyophilization conditions.

Commonly used protectant include saccharides, polyols, polymers, surfactants, some proteins and amino acids etc. We usually add 8% (mass ratio by volume) of trehalose and mannitol as lyoprotectant. Trehalose can significantly prevent the alter of the protein secondary structure, the extension and aggregation of proteins during freeze-drying process; mannitol is also a universal applied protectant and fillers, which can reduce the aggregation of certain proteins after lyophilization.

Our protein products do not contain carrier protein or other additives (such as bovine serum albumin (BSA), human serum albumin (HSA) and sucrose, etc., and when lyophilized with the solution with the lowest salt content, they often cannot form A white grid structure, but a small amount of protein is deposited in the tube during the freeze-drying process, forming a thin or invisible transparent protein layer.

Reminder: Before opening the tube cap, we recommend that you quickly centrifuge for 20-30 seconds in a small centrifuge, so that the protein attached to the tube cap or the tube wall can be aggregated at the bottom of the tube. Our quality control procedures ensure that each tube contains the correct amount of protein, and although sometimes you can't see the protein powder, the amount of protein in the tube is still very precise.

To learn more about how to properly dissolve the lyophilized recombinant protein, please visit Lyophilization FAQs.