Horseradish peroxidase (HRP) Rapid Coupling Kit

Catalog Number: UA080470 Brand: UA BIOSCIENCE

Price:

$370 USD

Size: 100μg 500μg 1mg

Quantity:

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Product Details

Product Specification

Synonyms

Rapid HRP Conjugation Kit

Background

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

Features and Benefits:
The kit provides a complete set of reagents required for rapid labeling, and the reagents are easy to store.
100% recovery rate, with a conjugation process completed in 2 hours and hands-on time of approximately 5 minutes.
The conjugated products require no purification and can be stored for extended periods.

Components

100μg can be used to label 100µg of antibody (monoclonal, polyclonal) or other proteins:

Component	Volume	Storage Temperature	Remarks
Reagent A	20µL	-20°C	Antibody/Protein Diluent
Reagent B	10µL	-20°C Store protected from light	Crosslinker
Reagent C	15µL	-20°C Store protected from light	Activated Enzyme

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

Component	Volume	Storage Temperature	Remarks
Reagent A	50µL	-20°C	Antibody/Protein Diluent
Reagent B	30µL	-20°C Store protected from light	Crosslinker
Reagent C	60µL	-20°C Store protected from light	Activated Enzyme

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

Component	Volume	Storage Temperature	Remarks
Reagent A	100µL	-20°C	Antibody/Protein Diluent
Reagent B	60µL	-20°C Store protected from light	Crosslinker
Reagent C	110µL	-20°C Store protected from light	Activated Enzyme

Protocol

100μg Usage Method:
1. Add Reagent A to the antibody or protein to be labeled (concentration range: 3-10mg/mL) (volume ratio of protein to Reagent A is 9:1), mix thoroughly by pipetting repeatedly, avoiding bubbles;
2. Continue by adding Reagent B (5µL) to the antibody or protein to be labeled, mix thoroughly by pipetting repeatedly; avoid bubbles;
3. Incubate at room temperature in the dark with gentle mixing for 1 hour;
4. Continue by adding Reagent C (10µL), mix thoroughly by pipetting repeatedly; avoid bubbles.
Note: Depending on the protein or antibody, the amount of Reagent C may be adjusted as needed.
5. Incubate at room temperature in the dark with gentle mixing for 1 hour.

500μg Usage Method
1. Add Reagent A to the antibody or protein to be labeled (concentration range: 3-10mg/mL) (volume ratio of protein to Reagent A is 9:1), mix thoroughly by pipetting repeatedly, avoiding bubbles;
2. Continue by adding Reagent B (25µL) to the antibody or protein to be labeled, mix thoroughly by pipetting repeatedly; avoid bubbles;
3. Incubate at room temperature in the dark with gentle mixing for 1 hour;
4. Continue by adding Reagent C (50µL), mix thoroughly by pipetting repeatedly; avoid bubbles.
Note: Depending on the protein or antibody, the amount of Reagent C may be adjusted as needed.
5. Incubate at room temperature in the dark with gentle mixing for 1 hour.

1mg Usage Method
1. Add Reagent A to the antibody or protein to be labeled (concentration range: 3-10mg/mL) (volume ratio of protein to Reagent A is 9:1), mix thoroughly by pipetting repeatedly, avoiding bubbles;
2. Continue by adding Reagent B (50µL) to the antibody or protein to be labeled, mix thoroughly by pipetting repeatedly; avoid bubbles;
3. Incubate at room temperature in the dark with gentle mixing for 1 hour;
4. Continue by adding Reagent C (100µL), mix thoroughly by pipetting repeatedly; avoid bubbles.
Note: Depending on the protein or antibody, the amount of Reagent C may be adjusted as needed.
5. Incubate at room temperature in the dark with gentle mixing for 1 hour.

Guidelines

1. This reagent is sensitive to moisture. Before opening the vial, ensure all component vials are fully equilibrated to room temperature to avoid moisture condensation inside the container. 2. Requirements for the antibody/protein to be conjugated: The antibody/protein should be stored in a 10 mM phosphate buffer (1x PBS, pH 7.2-7.4). The recommended concentration range is 3-10 mg/mL. The buffer must not contain any amine groups to avoid competitive reactions with the coupling reagent, which may affect conjugation efficiency. Specifically, avoid primary amines (e.g., Tris, glycine, ammonium salts, EDTA), protein stabilizers (e.g., BSA <0.1%, gelatin, glycerol <20%), and preservatives such as sodium azide. If the sample contains substances that may interfere with labeling, it is recommended to exchange the buffer with PBS and concentrate to the recommended concentration. 3. Repeated freeze-thaw cycles significantly impact this reagent. Any unused thawed reagent should be discarded and not reused. 4. Storage of conjugated products: If the final concentration of the antibody/protein conjugate is less than 0.5 mg/mL, consider concentrating or adding stabilizers. Store at 2-8°C with a shelf life of 3 months. For long-term storage, the conjugated product can be lyophilized or stored at -20°C after adding 50% glycerol.

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Horseradish peroxidase (HRP) Rapid Coupling Kit