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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
Can’t find the exact format you need? We can help you source or customize this human DNA polymerase alpha activity assay — including larger pack sizes, bulk reagents, bulk enzyme, or a protocol adapted to your samples. Click Talk to a Scientist, email support@biohippo.com, or explore our Research Services. Our team will be in touch shortly.