HRP Antibody Conjugation Check Kit (ab236555)
Overview
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Product name
HRP Antibody Conjugation Check Kit
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Product overview
HRP Antibody Conjugation Check Kit (ab236555) is a quick immunochromatography test that allows to confirm the successful conjugation of Horseradish Peroxidase (HRP) to an 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. Both Protein A and Protein G have a high affinity for the Fc region of a variety of IgG molecules. The half strips also contain an absorbent pad to promote and control the flow of sample through the nitrocellulose. 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 appears on the strip.
Benefits:
Quick and easy qualitative test to check success of conjugation
Sensitivity: the range of detection is between 0.5 ng/mL to 10 ng/mL.
Compatible with antibody conjugates generated with Lightning-Link® HRP conjugation kit (ab102890) or any other HRP conjugation technologies available commercially
No costly equipment required
Small sample volumes required (40 μL)
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Notes
This product is manufactured by Expedeon, an Abcam company. It was previously called HRP Check&Go! Antibody Labeling Confirmation Kit. Expedeon product code 4002-0030 is the same as the 30-test size of this product.
Properties
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Storage instructions
Please refer to protocols. -
Components 30 tests ab274295 - GAR-HRP (lyophilized) 1 vial ab274086 - HRP detection solution 1 x 1500µl ab274081 - Protein A/G Strips 1 x 30 tests ab274082 - Running Buffer (10X) 3 x 1500µl -
Research areas
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Relevance
Horseradish Peroxidase (HRP) is an enzyme commonly used as an indicator for chemical reactions which produce peroxide. The enzyme is routinely conjugated to antibodies for use in enzyme-based immunoassay systems. HRP functions in the removal of H2O2 (hydrogen peroxide), oxidation of toxic reductants, biosynthesis and degradation of lignin, suberization, auxin catabolism, response to environmental stresses such as wounding, pathogen attack and oxidative stress. These functions might be dependent on each isozyme/isoform in each plant tissue. -
Cellular localization
Secreted Probable. Vacuole Probable. Note: Carboxy-terminal extension appears to target the protein to vacuoles. -
Alternative names
- Horseradish Peroxidase
- HPRC1
- HRP
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Images
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How The Kit Works
The nitrocellulose membrane contains a Test line of immobilized Protein A and Protein G called a “half strip”. Both Protein A and Protein G have a high affinity for the Fc region of a variety of IgG molecules (see table “Protein A and Protein G affinity for immunoglobulins”). The half strips also contain an absorbent pad to promote and control the flow of sample through the nitrocellulose. 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 appears on the strip.
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HRP Antibody Conjugation Check KitGoat 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.
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HRP Antibody Conjugation Check KitIgG 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).