Lateral Flow Coupling Kit

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Application: Lateral Flow

Particles: Not included in this kit. This kit is for magnetic and non-magnetic latex and silica particles ranging in size from 200 nm to 500 nm

For Best Results: You will require a sonication bath to use this kit

Base: Water-based Activation Reagent is included in this kit

Product Highlights:

  • Demonstrated to increase the sensitivity of lateral flow assays and reduce variation in results

  • This kit enables scientist to bind any antibody without pre-treatment

  • Hand on time for surface activation and coupling is 60 minutes

  • An alternative to covalent NHS/EDC and passive binding the Anteo Kit is easy to use, while reducing damage to antibodies

  • Easy to use, you simply pipette the Activation Reagent on your surface to activate.

  • Promotes monolayer binding which increases antibody functionality

  • Reduces aggregation

  • Increases uniformity between experiments

Product Description

Reduce labour costs by using our ready-to-use Lateral Flow Coupling Kit for magnetic and latex particles. No messy pre-preparation of reagents required. Simply pipette Anteo's water-based Activation Reagent onto your chosen particles to activate. Hands-on time for surface activation and coupling is 60 minutes.

 

Magnetic particles are easier to use than gold. Magnetic separation removes the need to perform centrifugation and filtration concentration. This Lateral Flow Coupling Kit has successfully given scientists 5 x more sensitive lateral flow assays and less variation in their experiments than covalent EDAC chemistry when using magnetic particles.                       

 

This kit enables scientist to bind any antibody without pre-treatment. Promoting monolayer binding, this kit greatly reduces the risk of antibody damage, leading to less antibodies required for the experiment, increased antibody functionality, reduced interference and increased uniformity between experiments.

Frequently Asked Questions – Lateral Flow Coupling Kit
 

Q: Are particles provided in the kit?

A: No but this kit is for with magnetic and latex particles only. Not gold.

 

 

Q: Can I use this kit with gold particles?

A: No but we are developing an Anteo Gold Coupling Kit for Lateral Flow, sign up to our mailing list to hear when this kit will be available.

 

 

Q: How long does surface activation and coupling take with the Lateral Flow Coupling Kit?

A: Hands on time for surface activation and coupling is 60 minutes.

 

 

Q: What is different about Anteo’s Lateral Flow Kit?

A: The answer lies in Anteo’s Patented Technology:

 

1. Is very strong yet flexible, allowing the user to bind any antibody subclass without pre-treating the antibody and has been demonstrated to achieve 5x more sensitivity than the traditional covalent chemistry method.

 

2. Promotes monolayer binding: meaning antibodies assemble in the correct orientation reducing the amount of damage leading to increased functionality of antibodies. As a result, less antibody is required supporting a cost-efficient solution per sample.

 

3. Reduces aggregation: The activation reagent (included in the kit) contains a small amount of optimised surfactant, which reduces aggregation due to hydrophobic forces. It also creates a positively charged surface improving particle handling.

 

4. It is built upon chelation and coordination chemistry, which finally gives researchers the choice to use another method apart from passive and covalent bindings.

 

 

Q: What particles are compatible with this kit?

A: Lateral Flow Kit compatible with carboxylated magnetic, polystyrene and silica particles ranging from 200-500 nm. The following table shows tested particles:

 

ademtech Carboxyl-Adembeads 200nm

200nm

PS

Magnetic

ademtech Carboxyl-Adembeads 500nm

500 nm

PS

Magnetic

Allrun AllMag (PM3-020)

200 nm

PS

Magnetic

Merck Millipore Estapor (M1-020/50)

200 nm

PS

Magnetic

Merck Millipore PureProteome Carboxy FlexiBind

300 nm

PS

Magnetic

Microparticles Polystyrene particles (PS-MAG-COOH-S2632)

350 nm

PS

Magnetic

ThermoFisher Blue Colored Carboxylate-modified (DB1040CA)

400 nm

PS

Non-Magnetic

 

Q: Is the Lateral Flow Kit easy to use?

A: Yes. Simply pipette Anteo’s Activation Reagent onto your chosen surface. The reagent is water based and all of the buffers you need from surface activation to coupling, blocking and storage are provided. Hands on time for surface activation and coupling is 60 minutes.

 

 

Q: What type of linkage is formed?

A:  Our technology is built upon chelation and coordination chemistry with a binding strength equivalent to traditional covalent methods. The resulting activated surface functions as a double-sided velcro, whereby it strongly binds to your selected surface while allowing the gentle, yet strong binding of antibodies to the activated surface.

 

 

Q Can antibodies from different species be used?

A: Yes. Our Lateral Flow Kit has been tested with antibodies from a variety of species including mouse, rabbit.

 

Q: What proteins are compatible with this kit?

A range of proteins are compatible with this kit from Antibodies to Protein A. The kit has been tested with the following proteins: Mouse IgG, Mouse IgM, Rabbit IgG, Human IgG, Human IgA, Protein A, Protein G, Streptavidin.

 

The biggest advantage when using our kit is that you don't need to know anything about your antibody subclass to get accurate results and remove lengthy R&D steps before running your experiment. The flexibility to bind any antibody to your chosen bead without pre-treating your Antibody is finally here!

 

 

Q: Can monoclonal and polyclonal antibodies be used?

A: Yes. Our Lateral Flow Kit has been developed and tested for use with both monoclonal and polyclonal antibodies.


 

Q: What protein concentration should be used?

The Anteo protocol requires less antibody (at times using half the amount of antibody) to achieve 5 times more sensitivity that covalently conjugated bead based magnetic particles.

 

The suggested coupling range for this kit is 25 to 100 µg/mg of particles. The particles are activated and coupled at 10 mg/mL so this would mean that the final antibody concentration range is 250 µg/mL to 1,000 µg/mL. It is suggested that the optimum concentration is determined for your antibody.

 

 

Q: What should be done if aggregation is observed?

A: The activation reagent (included in the kit) contains a small amount of optimised surfactant which reduces aggregation due to hydrophobic forces. It also creates a positively charged surface improving particle handling.

 

In a case where aggregation is observed, vortex to resuspend and sonicate for 5 min. If aggregation is still observed repeat the process.  Sonication times and amount required will depend on type and power of sonication.  We recommend a bath sonicator with at least 60W power for 5 mins.  The user may have to optimise this step for their particles and equipment.  We also suggest a method of observing the aggregation has been dispersed, such as microscopy or DLS measurement.

 

 

Q: What sort of sonication can be used?

A: For the volumes in the IFU use a bath sonicator with fresh distilled and degassed water (minimum power 60W) For larger volumes a probe may be used. A probe sonicator would have to be optimised but general settings might be e.g.Sonics VC-750 Vibracell tip-sonicator 40% amplitude for 30 seconds.

 

 

Q: What other chemicals can be used during coupling?

Refer to chemical compatibility table in the instructions for use for information on what can and cannot be used during coupling. It is suggested to use the included Coupling Buffer to dilute the antibody and if the antibody cannot be diluted more than 25% then buffer exchange may be required.

 

 

Q: Do the particles require blocking after coupling and what blockers do you recommend?

It is highly recommended to block these particles using the Blocking Buffer included with the kit.  Other blockers may be assessed on an individual basis by addition to the provided Coupling buffer.


 

Q: What is the recommended buffer for drying the particles (conjugate) onto the conjugate pad?

The drying buffer should be optimised as this will be determined by the particle type and assay. For example when using 200 nm magnetic particles, we have used TBS buffer, pH8 containing 1% BSA, 5% Sucrose, 1% Tween20 and 0.05% Triton X-100.

 

 

Q: Can surfactants be used during the lateral flow assay (e.g. in the running buffer and conjugate drying buffer)?

Surfactants may be used during the assay and are recommended to reduce non specific binding. Refer to the chemical compatibility table in the instructions for information about surfactants during the activation and coupling steps.

 

 

Q: Are particles coupled with this kit compatible with fluorescent markers using in lateral flow?

Reduction in fluorescent signals has not been observed when using this kit and are therefore compatible.

 

 

Q: What reagents need to be avoided during storage and coupling?

It is recommended to use the included Coupling and Storage Buffer. For more information refer to chemical compatibility table in the instructions for use.

 

 

Q: Do I have to desalt the antibodies if they come in PBS?

If the antibody cannot be diluted more than 25% (final phosphate concentration ≤ 2.5 mM) then buffer exchange or desalting will be required.  Phosphate is a metal chelator and can interfere with the coupling process.

 

 

Q: If so, what method do you suggest?

There are a number of options to buffer exchange, desalt, or dialyse the antibody solution. Zeba™ Spin Desalting Columns (Thermo Fisher) work well without loss or dilution of the antibody. Use the included instructions for buffer exchange.

 

 

Q: What magnetic separators do you use?

The magnetic separator used during the development of this kit is the DynaMag-2 available from Life Technologies (Cat No. 12321D). Other magnets may be suitable as long as the supernatant achieves complete functional clearing.

 

 

Q: Can the particles be stored frozen for later use?

It is not recommended to freeze the particles as this can cause irreversible clumping or clustering. Repeat freezing and thawing particles may create cracks on the surface of the beads that result in release of iron from the beads and contamination of the sample.


 

Q: Can the activated particles be stored lyophilized (freeze dried) for later use?

After using our technology your activated particles can be lyophilised. A small number of magnetic microparticles have been lyophilised and subsequently resuspended and coupled including particles validated with this kit.


 

Q: How can the drying of particles be prevented?

Drying can be prevented by storing the particles in the supplied buffer in an airtight container at 2-8ºC.

 

 

Q: What is the stability of the activated particles?

The activated particles are stable for up to one year, when stored at 2-8ºC.

 

 

Q: What is the stability of antibody- coupled particles?

Stability of the antibody-coupled particles will depend on the antibody itself. Generally coupled particles are stable for up to one year, when stored in Storage Buffer at 2-8ºC.

 

 

Q: What is the stability of the particles dried on the conjugate pad?

Stability of the antibody coupled particles when dried will depend on the antibody itself and the dry buffer used and would have to be determined.

 

 

Q: Do the particles require blocking after coupling and what blockers do you recommend?

It is highly recommended to block these particles using the Blocking Buffer included with the kit.

 

 

Q: Do the particles require quenching after blocking/coupling?

As our technology uses metal chelation as opposed to active esters, like NHS/EDC, Tosyl or Epoxy, quenching is not required.

 

 

Q: What is the procedure for co-coupling two or more proteins to the particles?

A mixture of protein for co-coupling can be added in the same way as for a single antibody, within the coupling concentrations recommended to produce multifunctional particles, or mix with a blocker to reduce the amount and space out a coupled antibody, for example. As different types of protein will bind at varying rates, it is recommended to titrate all proteins, keeping the total concentration the same. See table below for example.  It is recommended to still block the particles after the coupling step.

 

[Antibody 1]

[Antibody 2]

[Total]

0 µg/mL

200 µg/mL

200 µg/mL

25 µg/mL

175 µg/mL

200 µg/mL

50 µg/mL

150 µg/mL

200 µg/mL

75 µg/mL

125 µg/mL

200 µg/mL

100 µg/mL

100 µg/mL

200 µg/mL

125 µg/mL

75 µg/mL

200 µg/mL

150 µg/mL

50 µg/mL

200 µg/mL

175 µg/mL

25 µg/mL

200 µg/mL

200 µg/mL

0 µg/mL

200 µg/mL

 

In This Kit:

  • Particle Pre-treatment Solution
  • Particle Activation Solution
  • Particle Intermediate Solution
  • Coupling Buffer
  • Blocking Buffer
  • Storage Buffer

Particles: Not included in this kit. This kit is for magnetic and latex particles

For Best Results: You will require a sonication bath to use this kit

Shipping: Ambient

Storage Temp: 2 – 8°C

Shelf life: 12 months from date of manufacture

SKU VMPLFMP
Brand Anteo
Shipping Weight 0.2500kg
Shipping Width 0.170m
Shipping Height 0.080m
Shipping Length 0.160m
Shipping Cubic 0.002176m3

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