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Home / Assay Kits / Drug Target Assays

Human DNA Polymerase Alpha Assay Kits

SKU : BHE16300001

This product includes all the assay kit components except the enzyme for 100 assays in a 384-well plate assay format: 400 ul of 10 x Buffer, 33 ul of 100 x DNA template, 33 ul of 100 x dNTP mix, 1550 ul of 2 x Dye, 1550 ul of 50 mM EDTA. The kit does not include the enzyme.

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Mfr.No.	HDPA100K, HDPA100KE1
Description	Human DNA Polymerase Alpha (Pol α) is responsible for the initiation of DNA replication at origins of replication The Pol α enzyme consists of four subunits: the catalytic subunit POLA1, the regulatory subunit POLA2, and the small and the large primase subunits PRIM1 and PRIM2. The human DNA polymerase alpha assay is based on measurement of the DNA molecules synthesized by the DNA polymerase catalytic subunit. The assay is performed in a 384-well or 96-well plate format. The assay can be used for detection of DNA polymerase alpha activity and high throughput screen of human DNA polymerase alpha inhibitors. The Human DNA Polymerase Alpha Assay Kit (Catalog No. HDPA100K) includes all the assay kit components except the enzyme for 100 assays in a 384-well plate assay format: 400 µl of 10 x Buffer, 33 µl of 100 x DNA template, 33 µl of 100 x dNTP mix, 1550 µl of 2 x Dye, 1550 µl of 50 mM EDTA. The kit does not include the enzyme.
Detection Method	/
Drug target assays	Anticancer drug targets

Product Manual	Datasheet
Protocols Description	Assay protocol The following assay protocol is based on the 384-well plate assay format (plate type: Matrix 4318 or alike). The reaction volume is 30 µl and the final assay volume is 60 µl. For 96-well plate assays (plate type: Costar 3915 or alike), the reaction volume is 60 µl and the final assay volume is 120 µl. 1. Reagent preparation: (1) 10 x DNA: Dilute the 100 x DNA 10-fold with water. Each assay uses 3 µl 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 3 µl of 10 x dNTP. (4) 1 x dye: Dilute the 2 x fluorescence dye 2-fold with 50 mM EDTA. Each assay uses 30 µl of 1 x dye. 2. Reaction: The total volume of each reaction mixture is 30 µl including 18 µl of H2O, 3 µl of 10 x Buffer, 3 µl of 10 x DNA template, 3 µl of 10 x enzyme, 3 µl 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: Mix 30 µl of the 1 x fluorescence dye with 30 µl 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.	Specifications	Size	Your price	Quantity
BHE16300001-1	HDPA100K	enzyme not included	100 assays	$378.19		Add to Cart
BHE16300001-2	HDPA100KE1	enzyme included	100 assays	$848.57		Add to Cart

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