Product Overview
EnkiLife HRP labeling kit contains multiple components for labeling antibody , such as the activated lyophilized HRP, labeling buffer, and other chemical reagents, enabling easy and reliable antibody labeling.
Product Features
- Activated HRP with extended arm:The cross-linker used in the kit has an extended arm to ensure that the application effect is not affected by protein steric hindrance.
- Site-specific conjugation:Optimized site-specific conjugation technology ensures uniform antibody labeling and improves efficiency.
- Rapid process:The simple procedure allows for efficient labeling completion in as fast as 3 - 4 hours.
- Highly active HRP：The HRP configured in this kit is of high purity and high activity, Enzyme activity is 120 - 200 U/mg，freeze-dried powder can be stably stored at -20°C for at least 12 months.
- User-friendly operation:Each HRP reagent is pre-optimized for corresponding antibody amounts. No complex calculations are required if the antibody sample meets the labeling conditions. The solid HRP ensures batch stability.
- Comprehensive reagents: The kit includes a labeling preservation solution, covering the entire process from labeling to preservation. No additional reagents are needed, making the process worry-free and convenient.
Kit Specifications
| Product Components | Component Content in Different Specifications | Storage Temperature | ||
|---|---|---|---|---|
| 80μg Antibody | 400μg Antibody | 4mg Antibody | ||
| Activated PerCP | 1 tube | 5 tube | 5 tubes | -20°C after opening, protected from light |
| Labeling Buffer H | 5 mL | 5 mL | 5 mL | 4 °C, away from light |
| Antibody Modification Reagents | 60 μL | 60 μL | 60 μL×2 | -20 °C, away from light |
| Ultrafiltration Tube * ,50K MWCO | 1 set ** | 1 set ** | 1 set ** | RT |
| Blocking reagent | 300 μg | 300 μg | 300 μg×4 | -20 °C, away from light |
| DMSO (dissolve blocking reagent) | 50 μL | 50 μL | 200 μL | -20 °C, away from light |
| Labeled protein storage solution | 400μL | 2 mL | 10 mL | 2-8 °C |
| Recommended amount of labeled antibody | Each tube dyes for labeling 20-80μg antibody Recommended: 20μg antibody | Each tube dyes for labeling 20-80μg antibody Recommended: 20μg antibody | Each tube dyes for labeling 200-800μg antibody Recommended: 200μg antibody | 2-8 °C |
* * Ultrafiltration tube molecular weight cutoff: 50KDa
** 1 set = 1 ultrafiltration tube and collection tube
Labeling Principle
This kit utilizes the cysteine characteristics of the antibody and the free amino groups on the dye to covalently couple the antibody molecule with the dye using a directed docking coupling technique.
Standard Labeling Procedure
- STEP 1：Ultrafiltration buffer exchange and concentration (optional)
- STEP 2：Antibody activation
- STEP 3：Labeling reaction
- STEP 4：Blocking and purification
- STEP 5：Storage
Data of HRP labeling kit application
Western Blot(WB) Analysis
Use the HRP labeling kit (RE80004p) to label two kinds of goat anti-mouse IgG antibodies, and then conduct WB detection to evaluate the effect.
- Sample: Mouse IgG, samples 1–6 are 500 ng, 50 ng, 5 ng, 500 ng, 50 ng, and 5 ng, respectively.
- Antibody:
HRP-labeled goat anti-mouse IgG 0.5 mg/ml (diluted 1:2000):
1–3: HRP-goat anti-mouse IgG (H+L)
4–6: HRP-goat anti-mouse IgG, F(ab)₂ fragment
Conclusion: The antibody staining showed clear signals with a expectant signal-to-noise ratio. RE80004P kit is suitable for labeling goat anti-mouse IgG.
Immunohistochemistry(IHC) Analysis
Use the HRP labeling kit (SKU: RE80004p), label goat anti-mouse IgG,then conduct IHC detection (DAB staining solution）to evaluate the effect.
- Sample: Human liver tissue paraffin sections
- Antibody:
Primary antibody: Vimentin mouse monoclonal antibody(diluted 1:100)
Secondary antibody:HRP-goat anti-mouse IgG preparaed with HRP labeling kit (SKU: RE80004p) with a 1:200 dilution
Conclusion: The Secondary antibody preparaed with HRP labeling kit staining showed clear signals with a expectant nuclear expression.
