$0.00
HRP Antibody Conjugation Kit
$300.00
Description
HRP Antibody Conjugation Kit offers an optimized process for rapid conjugation of horseradish peroxidase (HRP) to an antibody with high yields. The HRP Antibody Conjugation Kit includes all necessary reagents (modified HRP and antibody modifier). The conjugation efficiency of HRP to the antibody can reach up to 95%, while significant coupling is observed within one hour. Due to the high coupling efficiency, generally, purification of the product is not required. Thus, there is no loss of materials. In addition, the handling time is less than one minute.
HRP Antibody Conjugation Protocol
Advantages
•No loss of materials
•Less than one minute of handling time
•Very high coupling efficiency
•Modified HRP with high enzymatic activity (> 250 U/mg), ready for conjugation
•A long hydrophilic spacer (46 atoms) between antibody and HRP
•No homo-crosslinking
•Very stable and permanent conjugation
•Coupling at a pH range of pH7.0-9.0
•No reducing reagents
•No catalysts
Product specification
| Name | HRP Antibody Conjugation Kit |
| Modified HRP (D-HRP) | 3 vials |
| Antibody modifier | 3 vials |
| Recommended antibody | 3 x 100 μg (up to 3 x 400 μg) |
| Coupling group | Amines |
| Compatible pH | 7.0-8.0 |
| Compatible buffer | neutral PBS, HEPES, MES, MOPS |
| Application | In vitro research use only |
Delivery and storage
| Delivery | Cold packs |
| Short Term Storage | -20°C |
| Long Term Storage | -20°C |
| Other Notes | n/a |
Protocol
1. Bring the antibody modifier and modified HRP (D-HRP) to room temperature.
2. Prepare the antibody at 2.0 mg/ml in an appropriate buffer (pH 7.0-7.4), such as PBS or HEPES. Avoid BSA, sodium azide, and amine-containing reagents, such as glycine, tris, etc.
3. Add 1.0 ul antibody modifier to 50 ul (e.g. 100 ug) antibody solution and incubate for one hour at room temperature. Discard the residues of the antibody modifier. Flick the tube to mix the reaction every 15 minutes.
4. Add one vial of modified HRP (D-HRP) to the above reaction and incubate for 3 hours at room temperature. Frequently flick the tube to mix the reaction.
Significant conjugation can be observed within an hour, while a longer incubation period (up to overnight) could improve the coupling efficiency.
5. The conjugate is ready to use.
FAQ
1. How to prepare the labeling antibody?
The labeling antibody must be purified and have reactive amino (NH2) groups that are provided by lysine residues or the N-terminal amino acid. For optimum results, the antibody at 2.0 mg/ml should be prepared in an amine-free neutral buffer (PBS, MES, MOPS, HEPES, etc.). Sodium azide and protein additives (BSA, gelatin, etc.) should be avoided.
2. How much antibody can be conjugated in each reaction?
Each kit contains 3 vials of modified HRP (D-HRP) that is sufficient for 3 coupling reactions of 100 µg antibodies for the optimum result. However, up to 400 μg antibodies may be conjugated.
3. Is the purification of the conjugate required?
No. Although a small amount of unconjugated antibody and horseradish peroxidase may be present, in most applications, they do not interfere with the assay.
4. Can I use the HRP Antibody Conjugation Kit to conjugate a protein other than an antibody?
Yes. However, the coupling reaction may require to be optimized.
5. Can you make the HRP antibody conjugate for me?
Yes, we offer the custom HRP antibody conjugation service. Please click here to contact us or email us at in**@*******os.com.
