DNaseAlarm – Lateral Flow DNase Activity Assay Kit

DNaseAlarm Lateral Flow DNase Activity Assay Kit (catalog AU2056) is a DNase activity assay that shows whether active deoxyribonucleases are present in a liquid sample using a simple dipstick readout. Each kit includes 50 lateral flow dipsticks (4.5 mm) and a 96-well plate for running reactions. The workflow is a 20-minute reaction at 37 °C followed by a 15-minute strip run, and results are read by eye, by photograph or with a lateral flow reader.

The test uses a synthetic DNA substrate labeled with a 5′ biotin and a FAM tag. Intact substrate produces a visible test line; DNase activity cleaves the substrate and the test line fades or disappears.

Key Features

• Signal-loss readout: a strong test line means no DNase was detected; a weaker or missing test line indicates DNase activity.
• Defined positive response: according to the kit manual, the test line is expected to disappear at 2 units of DNase.
• Fast workflow: a 20-minute incubation at 37 °C and a 15-minute strip run.
• Kinetic evaluation: substrate cleavage increases over time when DNase is present, so results can be followed kinetically.
• Ready-to-use strips: pre-made dipsticks with an anti-FAM test line, so there is no need to stripe capture antibodies.
• Cost-effective: a low-cost way to screen for a DNase-free environment.
• Simple storage: the kit is stored at 2–30 °C; only the DNase substrate needs refrigeration.

What’s in the Kit

Each DNase activity assay kit includes:

• 50 lateral flow dipsticks, 4.5 mm
• DNaseAlarm Lateral Flow Running Buffer, 10 mL
• DNaseAlarm 1X Reaction Buffer, 10 mL
• DNase substrate containing biotin and FAM, 20 µL (refrigerate)
• One 96-well plate
• Instruction manual

You will also need: a timer, a pipettor and pipette tips (barrier tips are recommended), a way to incubate the plate at 37 °C, a centrifuge if specimens need to be clarified, and extra tubes or microtiter plates for running strips. A DNase I spike solution can be used to make positive controls.

Who It’s For

The DNaseAlarm kit suits:

• Molecular biology and genomics labs confirming that buffers, water and reagents are DNase-free
• Reagent and biologics manufacturers running quality control on production lots and in-process samples
• Enzyme groups analyzing the unit activity of DNase preparations or screening DNase inhibitors
• Core facilities that want a routine, low-cost screen for a DNase-free working environment

How to Use the DNase Activity Assay

1. Add 70 µL of DNaseAlarm Reaction Buffer to a well of the supplied 96-well plate.
2. Add the DNase substrate to the well as directed in the kit manual.
3. Add 5 µL of sample, negative control or positive control to each well. Run a positive and a negative control with each sample set so readings stay comparable across days, times and users.
4. Incubate for 20 minutes at 37 °C.
5. Add 70 µL of Running Buffer and mix well.
6. Place one lateral flow dipstick into each well and let it run for 15 minutes.
7. Read the strip by eye, by photograph or in a lateral flow reader.

The negative control should show both a test line and a control line within 15 minutes. Full instructions, storage conditions and precautions are in the DNaseAlarm kit manual (PDF). Use reagents from a single kit lot, as they are adjusted to work together.

How It Works and Reading Results

The DNA substrate carries a 5′ biotin that binds the streptavidin colloidal gold reporter, and a FAM label that is captured by the anti-FAM antibody at the test line. While the substrate is intact, it tethers gold to the test line and a visible line forms. When DNases are present, they degrade the substrate, the gold can no longer be held at the test line, and the signal is lost.

• Test line: the higher the DNase concentration in the sample, the lower the test line intensity compared with the negative control strip.
• Control line: the control line intensity may decrease as the test line intensity increases.
• Timing: read strips at the specified time. Line intensities can change as strips dry, so readings taken too early or too late may be inaccurate.

Frequently Asked Questions

What does a missing test line mean?

In this DNase activity assay, loss of the test line indicates that DNase in the sample cleaved the substrate. A strong test line, similar to the negative control, means no DNase activity was detected.

How sensitive is the kit?

The kit manual states that the test line is expected to disappear at 2 units of DNase. Below that level, the test line weakens as DNase concentration rises, so compare each strip with the negative control.

Can I use it to study DNase inhibitors?

Yes. The assay is intended for quality control testing and for analysis of the unit activities of DNase and DNase inhibitors. Because cleavage increases over time, results can also be evaluated kinetically.

What samples can be tested?

The DNaseAlarm DNase activity assay is designed for liquid samples such as buffers, reagents and other solutions. Use DNase-free solutions and reagents if you need to dilute samples.

How should the kit be stored?

Store the kit at 2–30 °C and keep the dipsticks in their sealed, desiccated container until use. Refrigerate the DNase substrate.

Do you offer related nuclease tests?

Yes. The RNaseAlarm Lateral Flow RNase Activity Assay Kit uses the same signal-loss format to detect ribonucleases, and the RNase H Lateral Flow Activity Assay Kit measures RNase H activity on an RNA/DNA hybrid substrate. For custom nucleic acid tests, see our nucleic acid lateral flow (NALF) test strips.

For research use only. Not for use in diagnostic procedures. Questions? Call 512-333-1330 or email sales@attogene.com.