Human DNA Polymerase Alpha Assay Kit

SKU:BHT16300001
Overview
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Fluorescence-based kit for measuring human DNA polymerase alpha activity and screening its inhibitors. Newly synthesized DNA is read at Ex 485 nm / Em 535 nm in 384-well or 96-well plates. Choose the Kit without enzyme or the Kit Plus with enzyme, each for 100 assays.
Assay Target Human DNA polymerase alpha activity
Detection Method Fluorescence
Detection Wavelength Ex 485 nm / Em 535 nm
Plate Format 384-well or 96-well plate
Assay Time About 65 min
Options selector
Catalog no. Format
HDPA100K Kit (enzyme not included)
HDPA100KE1 Kit Plus (enzyme included)
Available Options

Select the variant that best fits your experiment. Availability and lead time may vary by option.

  • Options: Format: Kit (enzyme not included), Kit Plus (enzyme included)
  • Lead time: options listed in "Availability Content"; other statuses may take longer.
  • Storage: Please contact us to confirm storage conditions.
  • Shipping: Shipped frozen.
  • Upon receipt: store at recommended temperature as soon as possible.
  • Sales terms and conditions: Please review prior to ordering.
Field Specification
Organism Human
Applications
  • Enzyme Activity
  • Compound Profiling
Assay type
  • Enzyme Activity Assay
  • Inhibitor Screening Assay
Assay target Human DNA polymerase alpha activity
Assay area Anticancer Drug Discovery
Target type DNA Polymerase
Detection method
  • Fluorescence
Detection wavelength Ex 485 nm / Em 535 nm
Instrument required Fluorescence microplate reader
Plate format 384-well or 96-well plate
Assay time
  • About 65 min
Storage Please contact us to confirm storage conditions.
Shipping Shipped frozen.
Catalog no. (Mfr.) HDPA100K | HDPA100KE1
Main SKU BHT16300001
Drug Target Assay Kit

Human DNA polymerase alpha (Pol α) starts DNA replication at replication origins. It is a four-subunit complex: POLA1 is the catalytic subunit, POLA2 is regulatory, and PRIM1 and PRIM2 form the small and large primase subunits. Pol α is an anticancer drug target.

Kit vs Kit Plus

Both formats contain reagents for 100 assays in a 384-well plate format. The only difference is the enzyme: the Kit (HDPA100K) has no enzyme, while the Kit Plus (HDPA100KE1) adds 30 µl of 100 x human DNA polymerase alpha (4 µM).

Kit (enzyme not included), 100 assays (HDPA100K) 100 assays
Kit Plus (enzyme included), 100 assays (HDPA100KE1) 100 assays

Kit Components

Component Kit (enzyme not included), 100 assays (HDPA100K) Kit Plus (enzyme included), 100 assays (HDPA100KE1)
10 x Buffer 400 µl 400 µl
100 x DNA template 33 µl 33 µl
100 x dNTP mix 33 µl 33 µl
2 x Dye 1550 µl 1550 µl
50 mM EDTA 1550 µl 1550 µl
100 x human DNA polymerase alpha (4 µM) Not included 30 µl

Assay Principle

The catalytic subunit copies a DNA template using dNTPs. After the reaction, a fluorescent DNA dye diluted in EDTA is added, and the fluorescence (Ex 485 nm / Em 535 nm) reflects the amount of newly made DNA, so polymerase inhibitors lower the signal.

Specifications

Assay Target Human DNA polymerase alpha activity
Target Type DNA Polymerase
Organism Human
Assay Type Enzyme Activity Assay; Inhibitor Screening Assay
Detection Method Fluorescence
Detection Wavelength Ex 485 nm / Em 535 nm
Plate Format 384-well or 96-well plate
Instrument Required Fluorescence microplate reader
Assay Time About 65 min (60 min reaction + 5 min detection)
Size 100 assays
Storage Please contact us to confirm storage conditions.
Shipping Shipped frozen.

Protocol

The manufacturer's protocol describes the procedure and reaction volumes.

  • Human DNA Polymerase Alpha Assay (PDF, see Documents): Covers catalog # HDPA100K, HDPA100KE1.

Safety & Handling

For Research Use Only (RUO). Not for use in diagnostic or therapeutic procedures. The Human DNA Polymerase Alpha Assay Kit is shipped frozen. Follow standard laboratory safety practices when handling the reagents. Please contact us to confirm storage conditions.
Q.What does this kit measure?
A.It measures the activity of human DNA polymerase alpha by detecting the DNA made by its catalytic subunit with a fluorescent dye. It is also used for high-throughput screening of Pol α inhibitors.
Q.What is the difference between the options?
A.Both formats contain reagents for 100 assays in a 384-well plate format. The only difference is the enzyme: the Kit (HDPA100K) has no enzyme, while the Kit Plus (HDPA100KE1) adds 30 µl of 100 x human DNA polymerase alpha (4 µM).
Q.Is a protocol available?
A.Yes. The manufacturer's protocol (Human DNA Polymerase Alpha Assay, linked under Documents) covers catalog # HDPA100K, HDPA100KE1.
Q.How long does the assay take?
A.About 65 min (60 min reaction + 5 min detection). See the protocol for the exact steps.
Q.How should the kit be stored?
A.The Human DNA Polymerase Alpha Assay Kit is shipped frozen. Please contact us for long-term storage conditions.

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