711-0001 Fluoro-Trap Fluorescein Labeling Kit

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1 kit / €260.00


Properties for Fluoro-Trap Fluorescein Labeling Kit

Product Category ELISA Kits
Quantity 1 kit
Shipping to Not USA/Canada
PDF datasheet View Datasheet
Manufacturer Novus Biologicals Inc.

Datasheet Extract

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This kit contains 3 x 100ug antibody.


2.1 Storage and components

The kit is shipped at ambient temperature in a tamper-evident polypropylene container. Store at 4C upon receipt.
Kit contents:
Three tubes of Fluorescein label
Three tubes of Fluoro-TrapTM resin
1 tube of Fluoro-TrapTM reaction buffer.
1 tube of Fluoro-TrapTM quencher
1 tube of Fluoro-TrapTM neutralizer
1 protocol

2.2 Considerations before use

2.2.1 Sample buffer
Ideally, the 100mg of antibody to be labeled should be in 10-50mM amine-free buffer, pH range 6.5 to 8.5. However, some aminecontaining buffers can be tolerated.

2.2.2 Sample volume
For the best results, the sample volume should be 50-200ml, which means that the concentration of antibody ideally should be 0.5mg/ml or more.

2.3 Setting up conjugation reactions

2.3.1. Before you add the solution of antibody to the Fluorescein label, add 2ml of Fluoro-Trap(TM) reaction buffer for each 10ml volume of antibody to be labeled. Mix gently.
2.3.2. Remove the screw cap from the vial of Fluorescein and pipette the antibody sample (with added Fluoro-Trap(TM) reaction buffer) directly onto the lyophilised material. Resuspend gently by withdrawing and re-dispensing the liquid a few times using a pipette.
2.3.3. Place the cap back on the vial and leave the vial standing for 3 hours at room temperature (20-25C).
2.3.4. After incubating for 3 hours, add 1ml of Fluoro-Trap(TM) Quencher for every 10ml of antibody used.
2.3.5. Incubate with the quencher for 1 hour (a longer period of incubation is okay).
2.3.6. Transfer the reaction mix to the tube insert which contains Fluoro-Trap(TM) resin. Agitate the contents periodically or place on an end-overend or other rotary mixing device. The Fluoro-Trap(TM) resin will turn orange as the free dye is absorbed from the solution of conjugate. This process is complete after 10 minutes.
2.3.6. Microfuge the collecting tube (with its tube insert) for ~20 seconds to collect the purified conjugate in the bottom of the collecting tube. The free dye remains associated with the Fluoro-Trap(TM) resin in the tube insert.
2.3.7. Discard the tube insert and spent resin and cap the collecting tube.
2.3.8. Add 2ul of neutralizing solution for each 10ul of conjugate.

2.4 Storage of conjugates

For any new conjugate, initial storage at 4C is the recommended option. A preservative may be desirable for long-term storage. Other storage conditions (e.g. frozen at -70C in small aliquots or stored in liquid form at -20oC with 50% glycerol) may also be satisfactory. The best conditions for any particular conjugate must be determined by experimentation.
Avoid buffer components that are nucleophilic, as these may react with the Fluorescein dye. Although Tris is an amine-containing buffer it has little effect on conjugation efficiency as long as the concentration is 50mM or less.
Azide causes significant interference and ideally should be removed or at least reduced in concentration to <0.01%.

Frequently asked questions

Q1. What functional groups do I need on my protein?
Fluoro-Trap(TM) systems requires amine groups on the biomolecule to be labeled. All antibodies have multiple amine functions.

Q2. Do I have to label 100mg of antibody?

No. The amount does not have to be exactly 100mg but this quantity gives a molar ratio of antibody to dye that leads to good labeling for most antibodies. Varying the amount of antibody using a fixed amount of dye (as provided in the kit) can be used to generate conjugates with a varying average number of dye molecules per antibody. Experimentation in the range 50-200mg antibody per tube may be useful in fine tuning the performance of conjugates for specific applications.

Storage The kit is shipped at ambient temperature in a tamper-evident polypropylene container. Store at -20C upon receipt.
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