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Horseradish peroxidase (HRP) Rapid Coupling Kit

Horseradish peroxidase (HRP) Rapid Coupling Kit

Catalog Number: UA080470 Brand: UA BIOSCIENCE
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Regular price $440 USD
Regular price Sale price $440 USD
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Product Details

Product Specification


Synonyms Horseradish Peroxidase (HRP) Rapid Conjugation Kit

Background

This kit provides a simple and rapid conjugation protocol, specifically designed for conjugating horseradish peroxidase (HRP) to antibodies, proteins, peptides, or other ligands containing free amino groups. Compared with traditional methods, our conjugation reaction offers higher efficiency and yield, and the conjugates are more stable and less likely to dissociate. The reagent reacts effectively with primary amino groups (-NH₂) in buffer solutions with a pH of 7-9, forming stable amide bonds. Proteins, including antibodies, typically have several primary amines on the side chains of lysine (K) residues and at the N-terminus of each polypeptide, which can serve as targets for reagent labeling.

Functions and Features:
This kit provides a complete set of reagents required for rapid labeling, and the reagents are easy to store.

100% recovery rate, coupling process takes 2 hours, and hands-on time is about 5 minutes.

The coupling product does not require purification and can be stored for a long time.

Components

100 μg is sufficient for labeling 100 µg of antibody (monoclonal, polyclonal) or other proteins.

Components

Volume

Storage temperature

Remark

Reagent A

20µL

-20°C

Antibody/Protein Diluent

Reagent B

10µL

 --20°C store in the dark

coupling agent

Reagent C

15µL

Store at 20°C away from light

activase

500μg can be used to label 500µg of antibodies (monoclonal antibodies, polyclonal antibodies) or other proteins:

Component

loading capacity

Storage temperature

Note

Reagent A

50µL

-20°C

Antibody/Protein Diluent

Reagent B

30µL

Store at -20°C away from light

coupling agent

Reagent C

60µL

Store at -20°C away from light

activase

1mg can be used to label 1mg of antibodies (monoclonal antibodies, polyclonal antibodies) or other proteins


component

loading capacity

Storage temperature

Note

 Reagent A

100µL

-20°C

Antibody/Protein Diluent

Reagent B

60µL

Store at -20°C away from light

coupling agent

Reagent C

110µL

Store at -20°C away from light

activase


Protocol

Usage of 100μg:
1. Add Reagent A to the antibody or protein to be labeled (with a concentration range of 3-10mg/mL) at a volume ratio of 9:1 between the protein to be labeled and Reagent A. Pipette repeatedly to mix well and avoid generating bubbles.


2. Continue to add 5µL of Reagent B to the antibody or protein to be labeled. Pipette repeatedly to mix well and avoid generating bubbles.


3. Mix and react at room temperature in the dark for 1 hour.


4. Continue to add 10µL of Reagent C. Pipette repeatedly to mix well and avoid generating bubbles.


Note: Due to differences in proteins or antibodies, the amount of Reagent C added can be appropriately adjusted according to specific circumstances.


5. Mix and react at room temperature in the dark for 1 hour.

Usage Instructions for 500μg
1. Add Reagent A to the antibody or protein to be labeled (with a concentration range of 3-10mg/mL) at a volume ratio of 9:1 between the protein to be labeled and Reagent A. Pipette repeatedly to mix well, avoiding the generation of bubbles.


2. Continue to add 25µL of Reagent B to the antibody or protein to be labeled, and pipette repeatedly to mix well; avoid generating bubbles.


3. Mix and react at room temperature in the dark for 1 hour.


4. Continue to add 50µL of Reagent C, and pipette repeatedly to mix well; avoid generating bubbles.


Note: Due to differences in proteins or antibodies, the amount of Reagent C added can also be appropriately adjusted according to specific circumstances.


5. Mix and react at room temperature in the dark for 1 hour.

Usage of 1mg
1. Add Reagent A to the antibody or protein to be labeled (with a concentration range of 3-10mg/mL) at a volume ratio of 9:1 between the protein to be labeled and Reagent A. Pipette repeatedly to mix well, avoiding the generation of bubbles.


2. Continue to add 50µL of Reagent B to the antibody or protein to be labeled. Pipette repeatedly to mix well, avoiding the generation of bubbles.


3. Mix and react at room temperature in the dark for 1 hour.


4. Continue to add 100µL of Reagent C. Pipette repeatedly to mix well, avoiding the generation of bubbles.


Note: Due to differences in proteins or antibodies, the amount of Reagent C added can also be appropriately adjusted according to specific circumstances.


5. Mix and react at room temperature in the dark for 1 hour.

Guidelines

1.This reagent is sensitive to moisture. Before opening the bottle, all component vials must be fully equilibrated to room temperature to prevent moisture condensation inside the container.

2. Requirements for the buffer and concentration of the antibody/protein to be coupled: The antibody/protein should be stored in 10mM phosphate buffer (1xPBS, pH 7.2-7.4) with a recommended concentration range of 3-10mg/mL. The buffer must not contain amino components to avoid competitive reactions with the coupling agent, which would affect the coupling effect. Specifically, it includes primary amines (such as Tris, glycine, ammonium salts, EDTA), protein stabilizers (such as BSA should be <0.1%, gelatin, glycerol should be <20%), and preservatives such as sodium azide. If the sample contains substances that may interfere with labeling, it is recommended to replace the buffer with PBS and concentrate to the recommended concentration.


3. Repeated freezing and thawing have a significant impact on this reagent. Any unused thawed reagent must be discarded and cannot be reused.


4. Storage of coupled products: If the final concentration of the antibody/protein label is less than 0.5mg/mL, consider concentrating or adding a stabilizer, and store at 2-8°C with a validity period of 3 months. For long-term storage, the coupled product can be lyophilized or stored at -20°C after adding 50% glycerol.