Product Overview
Biotin conjugation/ labeling in <20 mins with 30 secs hands-on time using Biotinylation Kit/ Biotin Conjugation Kit. Designed for full antibody recovery (100%).
Kit Components
| 1 mg | Quencher reagent (1 x 200 µL); Biotin Conjugation Mix (1 x 1 mg); Modifier reagent (1 x 200 µL) |
|---|---|
| 100 µg | Biotin Conjugation Mix (1 x 100 µg); Modifier reagent (1 x 200 µL) |
| 3 x 10 µg | Biotin Conjugation Mix (3 x 10 µg); Modifier reagent (1 x 200 µL); Quencher reagent (1 x 200 µL) |
| 3 x 100 µg | Quencher reagent (1 x 200 µL); Biotin Conjugation Mix (3 x 100 µg); Modifier reagent (1 x 200 µL) |
Storage
| Shipping | Ambient – Cannot Ship with Ice |
|---|---|
| Short-term storage | -20ºC |
| Long-term storage | -20ºC |
Applications and Use
Biotinylation Kit/ Biotin Conjugation Kit uses a simple and quick process for biotinylation/ biotin labeling of antibodies. It can also be used to conjugate other proteins or peptides.
Add modifier to antibody and incubate for 15 mins. Add quencher and incubate for 5 mins. The conjugated antibody can be used immediately in WB, ELISA, IHC etc. No further purification is required and 100% of the antibody is recovered for use.
The Biotinylation Kit/ Biotin Conjugation Kit is optimized to produce conjugates for assays in which the biotinylated protein is captured by streptavidin immobilized on a surface (e.g., plates, nitrocellulose, magnetic beads etc). Custom size conjugation kits up to 100 mg are available on demand.
Amount and volume of antibody for conjugation to Biotin
| Kit Size | Recommended amount | Maximum amount | Maximum antibody volume |
|---|---|---|---|
| 3 x 10 μg | 3 x 10 μg | 3 x 20 μg | 3 x 10 μL |
| 100 μg | 1 x 100 μg | 1 x 200 μg | 1 x 100 μL |
| 3 x 100 μg | 3 x 100 μg | 3 x 200 μg | 3 x 100 μL |
| 1 x 1 mg | 1 x 1 mg | 1 x 2 mg | 1 x 1 mL |
Using the maximum amount of antibody may result in less labelling per antibody. Ideal antibody concentration is 1 mg/ml. 0.5 – 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 2 mg/ml or < 0.5 mg/ml should be diluted /concentrated.
Buffer Requirements for Conjugation
Buffer should be pH 6.5-8.5. If a concentration is shown, then the constituent should be no more than the concentration shown. If several constituents are close to the limit of acceptable concentration, then this can inhibit conjugation.
| Compatible constituents | 50mM / 0.6% Tris; 0.1% BSA; 50% glycerol; 0.1% sodium azide; PBS; potassium phosphate; sodium chloride; HEPES; sucrose; sodium citrate; EDTA; trehalose. |
|---|---|
| Incompatible constituents | Thiomerosal; Proclin; Glycine; Arginine; Glutathione; DTT. |
Tris buffered saline is almost always ≤ 50 mM / 0.6%. BSA can also interfere with the use of the conjugated antibody in tissue staining. If a constituent of the buffer containing your antibody or protein is not listed above, please contact us. Only purified antibodies are suitable for use, i.e. where other proteins, peptides, or amino acids are not present: antibodies in ascites fluid, serum or hybridoma culture media are incompatible.
Storing and handling conjugation kits
Lyophilized components are hygroscopic. Kits are intentionally shipped at ambient temperature with silica gel to avoid exposure to moisture. Upon receipt, store the kit frozen and protect from moisture. Before opening the outer container, allow the lyophilized components to reach room temperature to minimize condensation.
For research use only, not for clinical diagnosis.
