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Product Overview
The 96-Well DNA-Binding Plates are polystyrene plates coated with positive charges that bind nucleic acids (DNA or RNA). They are used to study specific interactions between a bound nucleic acid and a second molecule, either another nucleic acid or a protein.
The nucleic acid is first bound to the plate through charge-charge interactions. After blocking, the plate is incubated with the second molecule, and any second molecule that binds to the immobilized nucleic acid is detected by biochemical or biophysical methods. The 96-well format can also be used to screen libraries of nucleic acids or proteins for binding to the immobilized nucleic acid.
Each pack contains 4 plates. Black plates give a low background for fluorescence detection. Transparent plates are for detection by light absorbance.
Example data: binding signal on a DNA-binding plate (left) and a protein-binding plate (right).
Product Description
| Option | Black (fluorescence detection) | Transparent (absorbance detection) |
|---|---|---|
| Catalog # | NBP96B | NBP96T |
| Plate Color | Black | Transparent |
| Detection | Fluorescence | Light absorbance |
| Contents | 4 plates | 4 plates |
Specifications
| Format | 96-well plates |
|---|---|
| Plate Color | Black or transparent (two options; see Product Description) |
| Pack Size | 4 plates per pack |
| Material | Polystyrene |
| Surface | Positively charged; binds DNA and RNA |
| DNA Binding Capacity | About 1.1 ng/mm² (measured with a 1,000-nt DNA oligo at 2 µg/µl) |
| Stability of Bound DNA | Stable below 200 mM salt, and with non-ionic detergents such as Triton X-100 and Brij-35 below their CMC; SDS interferes with DNA binding |
| Catalog # | NBP96B, NBP96T |
| Storage | Please contact us to confirm storage conditions. |
| Shipping | Shipped at room temperature. |
Protocol Outline
- Add 200 µl of 10 µg/ml DNA in a low-salt buffer (10 mM buffer, 10 mM NaCl, 0.01% Tween-20, 1 mM EDTA) to each well and incubate at 4°C for 2 hours to overnight; overnight binding gives more complete binding.
- Rinse, then block with 300 µl of blocker for 30 min: 1% BSA when the second molecule is a protein, or 1× Nucleic Acid Binding Blocker when it is a nucleic acid.
- Add 200 µl of the second molecule in low-salt buffer and incubate for 1 hour, then rinse.
- Detect the bound second molecule, for example by enzyme activity, a fluorescent or radioactive label, or SYBR Green fluorescence (Ex 485 nm / Em 535 nm) for nucleic acids.
The full ProFoldin protocol “96-well DNA–binding plates” is linked under Documents.
Notes
- The Nucleic Acid Binding Blocker used for nucleic acid partners is sold separately.
- The second molecule concentration depends on its binding affinity; about 0.1 mg/ml is typical for proteins, lower for antibodies, and micromolar for DNA oligos.
- Detergents may affect protein function in the binding step.
- When the second molecule is a nucleic acid detected with SYBR Green, include wells with the bound DNA alone as background, because the dye also stains the immobilized DNA.
Safety & Handling
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Arora S, Kothandapani A, Tillison K, Kalman-Maltese V, Patrick SM (2010) Downregulation of XPF-ERCC1 enhances cisplatin efficacy in cancer cells. DNA Repair (Amst), 9(7):745-753. 10.1016/j.dnarep.2010.03.010
Sawant A, Kothandapani A, Zhitkovich A, Sobol RW, Patrick SM (2015) Role of mismatch repair proteins in the processing of cisplatin interstrand cross-links. DNA Repair (Amst), 35:126-136. 10.1016/j.dnarep.2015.10.003
