HRP Check&Go! is a quick immunochromatography test that allows you to confirm the successful conjugation of Horseradish Peroxidase (HRP) to your antibody without the need for any specialized or costly equipment.
The key component of the kit is a nitrocellulose membrane containing a Test line of immobilized Protein A and Protein G called a half strip. The HRP-antibody conjugate is run on the Protein A/G Strip. The conjugate binds the Protein A and Protein G concentrated on the Test line.
After the addition of the HRP detection solution, a visible line on the strip appears.
Features and benefits:
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- Quickly check the success of your conjugation (only 25 minutes!)
- Easy to use
- Sensitive – range of detection is between 0.5ng/ml-10ng/ml
- No costly equipment required
- Compatible with antibody conjugates generated using our Lightning-Link® HRP antibody labeling kits, or any other HRP conjugation technologies available commercially
- Requires only small volumes (40ul of diluted conjugate)
- Unique to Expedeon
Figure 1. Goat anti-rabbit IgG was conjugated with Lightning-Link® HRP and run at different concentrations on Protein A/G strips. After the addition of HRP Detection Solution, a blue signal developed at the T-line. The range of detection to obtain a clear, strong result is from 0.5ng/ml to 10ng/ml of antibody conjugate.
Figure 2. IgG antibodies from mouse, rabbit and goat were conjugated with Lightning-Link® HRP and run at different concentrations on Protein A/G strips. After the addition of HRP Detection Solution, a blue signal developed at the T-line. There is a strong visible signal for the IgG of all three species within the suggested range (0.5ng/ml to 10ng/ml of antibody-conjugate).
1.What are the best storage conditions for the kit?
The kit is shipped at ambient temperature and its components need different storage temperatures. Upon receipt, the plastic pot containing the strips and the HRP detection solution should be stored at 4⁰C. The running buffer and the lyophilized control should be stored at -20⁰C.
The lyophilized control vial must be stored at -20⁰C; once reconstituted with dH2O, the GAR-HRP conjugate is stable at 4⁰C for up to 1 month.
2. Can antibodies from different species be used?
Yes, the HRP Check&Go! strips have been tested with antibodies from a variety of species including mouse, rabbit and goat, providing they are conjugated to HRP. The binding properties of Protein A and G, immobilized on the Test line of the strips, differ among species and subclasses of IgG. A low or no affinity for a specific IgG subclass may lead to low/no signal but the conjugate may be still fine.
3. Can conjugates from antibody fragments be checked?
The protein A and protein G immobilized on the Test line have a high affinity for the Fc region of IgG molecules, and therefore for the Fc fragments as well.
4. Can I use HRP Check&Go to confirm successful conjugation of my antibody to other labels?
HRP Check&Go! is only suitable for testing successful conjugation of your antibody to HRP, however we also provide Biotin Check&Go! for verifying successful biotinylation, and Conjugate Check&Go! for testing the success of conjugation to colored labels, such as fluorescent dyes and proteins, latex beads and gold nanoparticles.
Please see here for ordering & shipping information.