Product Information
Fluorescently-labeled antibodies can be used for a variety of immunoassays such as immunohistochemistry, immunocytochemistry, flow cytometry, and fluorescent Western blot.
This antibody conjugation kit is designed to covalently conjugate FITC Plus to antibodies in 1-2 hours. Conjugated antibodies can be used immediately or stored at 4°C for up to 12 months. This kit is suitable for three reactions of labeling 100 ug primary antibody.
Antibody requirements: This kit is compatible with IgGs at 1 mg/mL in carrier-free buffer (PBS) with <0.1% BSA. Antibody buffer must be azide and glycerol-free.
Components
Component | Catalog No. | Size |
Vial A: Antibody Modifier Reagent | PK3003X-A | 3*100 ug |
Vial B: Lyophilized FITC Plus | PK30032-B | 3*100 ug |
Storage Vial C | PK3003X-C | 3*100 ug |
Quenching Reagent | PK3003X-Q | 200 uL |
Storage
Store at room temperature (20-25℃) in the dark.
Usage
Note: Before you begin, equilibrate primary antibody to room temperature. Primary antibody should be in carrier-free buffer (PBS) at 1 mg/mL with <0.1% BSA. Spin down Vial A (Antibody Modifier Reagent) and Vial B (lyophilized dye) to ensure that lyophilized contents are at the bottom of the tube.
Protocol:
1. Add 100 ug of primary antibody to Vial A (lyophilized Antibody Modifier Reagent). Pipette to mix.
2. Incubate for 15 min at RT.
3. Transfer entire contents of Vial A to Vial B (lyophilized dye). Pipette to mix.
4. Rotate for 30 minutes to 1 hour at RT. Protect from light. Spin down Vial B.
5. Add 20 uL of Quenching Reagent to Vial B. Transfer entire contents of Vial B to Storage Vial C. Pipette to mix.
6. Incubate for 10 minutes. Conjugated antibody is now ready to use.
Picture Protocol:
Additional Information
1. Titrations of the conjugated antibody are recommended to find the optimal antibody dilution for your application.
2. Store conjugated antibody in Vial C sealed at 4°C. Blank labels are provided to label Vial C.
3. It is recommended to add a preservative such as sodium azide to the conjugated antibody for long-term storage.
4. The estimated final concentration of the conjugated antibody is 0.83 mg/mL.
Cited in Article as
pk30032, ChemBright FITC Plus Antibody Conjugation Kit, Proteintech, IL, USA