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Home / Enzymes, Inhibitors & Substrates / Human Enzymes

Human DNA Polymerase Alpha

SKU : BHE16300226

This product includes 7 ul of 4 uM human DNA polymerase alpha catalytic subunit.

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Mfr.No.	HDPA1-100, HDPA1-1000
Description	Protein construct: Wild-type human DNA polymerase alpha catalytic subunit purified from a baculovirus insect protein expression system. MW: 170 kDa Enzyme concentration: 4 µM DNA polymerase assay: The DNA polymerase beta activity is measured by using the Human DNA Polymerase Assay Alpha kit (Catalog No. HDPA100K). Storage temperature: -20 or -80°C. Do not freeze-and-thaw repeatedly. Enzyme dilution: Use the 1 x assay to dilute the enzyme just before the assay. Do not store diluted enzyme solution. Human DNA Polymerase Alpha – for 100 assays (Catalog No: HDPA1-100 ) includes 7 l of 4 µM human DNA polymerase alpha catalytic subunit. It is sufficient for 100 assays.

Product Manual	Datasheet
Protocols Description	Assay Protocol using Human DNA Polymerase Alpha Assay kit (Catalog No. HDPA100K) The following assay protocol is based on the 384-well plate assay format (plate type: Matrix 4318 or alike). The reaction volume is 30µl and the final assay volume is 60µl. For 96-well plate assays (plate type: Costar 3915 or alike), the reaction volume is 60µl and the final assay volume is 120µl. 1. Reagent preparation: (1) 10 x DNA: Dilute the 100 x DNA 10-fold with water. Each assay uses 3µl of 10 x DNA. (2) 10 x enzyme: The recommended final enzyme concentration is 4 to 8 nM in the assay. (3) 10 x dNTP: Dilute the 100 x dNTP (10 mM) 10-fold with water. Each assay uses 3µl of 10 x dNTP. (4) 1 x dye: Dilute the 2 x fluorescence dye 2-fold with 50 mM EDTA. Each assay uses 30µl of 1 x dye. 2. Reaction: The total volume of each reaction mixture is 30µl including 18µl of H2O, 3µl of 10 x Buffer, 3µl of 10 x DNA template, 3µl of 10 x enzyme, 3µl of 10 x dNTP. Incubate the reaction mixture at 37°C for 60 min. Note: The enzyme reaction rate will be slower if the assay is at a lower temperature. 3. Detection: 30µl of the 1 x fluorescence dye with 30µl of the reaction mixture for 5 min. Measure the fluorescence intensity at 535 nm using the excitation wavelength at 485 nm.

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Combination SKU	Supplier No.	Size	Your price	Quantity
BHE16300226-1	HDPA1-100	100 assays	$503.79		Add to Cart
BHE16300226-2	HDPA1-1000	1000 assays	$2123.68		Add to Cart

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