The Attogene Fluorescent Universal Lateral Flow Assay Kit (Quantum Dots), AU2037, is a ready-to-use fluorescent lateral flow assay kit for quickly and cost-effectively developing dipstick assays that detect labeled DNA and RNA. Each kit provides 50 pre-made 4.5 mm dipsticks with a streptavidin quantum dot conjugate pad, 10 mL of nuclease-free running buffer, labeled control nucleic acid and a 96-well plate.
The fluorescent particles have a broad UV excitation range (100 nm to 400 nm) and emit at 655 nm, so results can be read by eye with an ordinary black light (UV flashlight) or quantified in a fluorescent lateral flow reader. Fluorescent lateral flow assays have been described as 2 to 100 times more sensitive than gold-based tests. Most runs are complete in 10 to 15 minutes.
Key Features
- Universal format: Detects amplified DNA products from PCR, LAMP and RPA reactions, or any nucleic acid carrying biotin plus a second label.
- No striping required: Capture lines are already sprayed on the strips, so there is no need to stripe capture antibodies.
- Simple readout: Read with a black light by eye, or use a fluorescent lateral flow reader for numerical results and the highest sensitivity.
- Multiplex options: Available in 2-line, 3-line and 4-line configurations for single or multiplex detection.
- Fast: Results in about 10 to 15 minutes.
- Cost-effective screening: A low-cost way to screen designs before downstream lateral flow assay development.
What’s in the Fluorescent Lateral Flow Assay Kit
Each fluorescent lateral flow assay kit includes:
- 50 fluorescent lateral flow dipsticks, 4.5 mm (store at room temperature)
- 10 mL Nucleic Acid Lateral Flow Running Buffer, nuclease-free (room temperature)
- 20 µL control nucleic acid containing biotin and FAM (all versions; refrigerate or freeze)
- 20 µL control nucleic acid containing biotin and DIG (3-line and 4-line versions)
- 20 µL control nucleic acid containing biotin and DNP (4-line version only)
- One 96-well plate and manual
Store the kit at 2 °C to 30 °C and keep strips in the sealed foil pouch until use.
You will also need: a timer, a centrifuge for preparing clear specimens, a pipettor and tips, tubes or microtiter plates, and your molecule of interest labeled with biotin plus FITC/FAM, DIG or DNP (depending on version).
Line Configurations
- 2-line (01 version): Test line anti-FITC/FAM; control line biotin. Requires a biotin- and FAM-labeled product.
- 3-line (02 version): Test line 1 anti-FITC/FAM, test line 2 anti-DIG, control line biotin.
- 4-line (03 version): Test line 1 anti-DNP, test line 2 anti-FITC/FAM, test line 3 anti-DIG, control line biotin.
When generating products by amplification, design the primers so the final product carries both labels.
Who It’s For
This kit is designed for researchers and assay developers who want to move a nucleic acid amplification workflow onto a fluorescent lateral flow readout without investing in striping or cutting equipment. It suits groups evaluating isothermal (LAMP, RPA) or PCR-based detection, teams comparing fluorescent and colloidal gold formats, and developers screening primer and label combinations before building a custom test.
How to Use the Fluorescent Lateral Flow Assay Kit
- Add 150 µL of Nucleic Acid Lateral Flow Running Buffer to a well of the 96-well plate.
- Set up sample, positive and negative wells. For samples, add a designated amount of product (1 to 5 µL is a good starting point; try large dilutions on a first run and keep the volume below 15 µL if possible). For the positive control, add 5 µL of control oligonucleotide. Leave the negative control well with buffer only.
- Mix each well completely by pipetting up and down several times.
- Add one dipstick to each well, arrows facing up.
- Incubate for 10 to 15 minutes.
- Analyze by eye under a black light, or read in a fluorescent lateral flow reader.
Full instructions are in the AU2037 kit manual (PDF). For background data on detection limits and particle-to-oligo ratios, see the Attogene technical article (PDF).
Reading Results and Optimizing Your Assay
Labeled nucleic acid binds the streptavidin quantum dots and is captured at the matching test line. The higher the concentration of labeled product, the brighter the test line compared with the buffer-only strip. Free streptavidin particles bind the biotin control line, so control line intensity decreases as the test line increases. This makes the strip useful for ratio analysis.
The stoichiometry between labeled nucleic acid and particles matters. If no test line appears, a serial dilution may be needed because of the hook effect. If you need higher analytical sensitivity, increase the volume or concentration of nucleic acid added. Positive samples can be confirmed by agarose or acrylamide gel analysis.
Prefer a visible colorimetric readout? See the gold Universal Lateral Flow Assay Kit. Other fluorescent labels are available in the europium version and the R-phycoerythrin version.
Frequently Asked Questions
What labels does my nucleic acid need?
Every product needs biotin plus a second label: FITC/FAM for the test line in all versions, DIG for the 3-line and 4-line versions, and DNP for the 4-line version.
Do I need a special reader?
No. This fluorescent lateral flow assay kit can be read by eye with a standard black light. A fluorescent lateral flow reader gives numerical readings and is needed to reach the 2 to 100 times sensitivity gains described in the literature.
How long does a run take?
Generally 10 to 15 minutes. The positive control should give test and control line signal within 15 minutes.
Why is my control line getting weaker?
That is expected. As more labeled product binds the particles, fewer free particles reach the biotin control line.
Can the kit be customized?
Yes. The fluorescent lateral flow assay kit can be configured with alternative or custom streptavidin fluorescent particles and other line setups. Attogene also offers custom lateral flow assay development services.
For research use only. Not for use in diagnostic procedures. Questions? Call 512-333-1330 or email sales@attogene.com.
