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Invitrogen™ pHrodo™ Antibody Labeling Kits
Description
Elevate biopharmaceutical research with pHrodo Antibody Labeling kits, designed to characterize antibody internalization, a critical step during antibody-drug conjugate research and development. Kits provide a simple, detailed protocol and advanced purification columns to easily label and purify antibodies and other large proteins. Amine-reactive pHrodo dyes covalently bind to antibodies and other proteins to produce bright, soluble conjugates whose fluorescence increases in acidic organelles including endosomes and lysosomes. Antibody internalization can be evaluated using flow cytometry, fluorescence microscopy, and high-content screening.
The pHrodo Red Antibody Labeling Kit and pHrodo Green Antibody Labeling Kit provide the reagents needed to easily label three purified protein or antibody samples with pHrodo Red or pHrodo Green dye. These kits contain amine-reactive, pH-sensitive pHrodo Red or Green STP ester dye that covalently binds to free lysines anywhere they occur in an antibody or protein.
The dyes included in this kit are improved versions of pHrodo Red and pHrodo Green dyes. They are optimized for the creation of bright and soluble bioconjugates that are used in the study of antibody internalization, endocytosis, phagocytosis, and for development of antibody-drug conjugates. These dyes are also more soluble than the original pHrodo Red and pHrodo Green dyes, making them useful for the labeling of antibodies that may otherwise precipitate out of solution during conjugation. pHrodo Red and pHrodo Green dyes are non-fluorescent outside the cell, but fluoresce bright red or bright green as the pH of their surroundings becomes more acidic, such as those found inside of phagosomes and endosomes.
The pHrodo Red and Green Microscale Labeling Kits offer the following benefits:
• Convenience—Labeling typically takes one hour, with less than 30 minutes of hands-on time
• Versatility—Labelling can be performed on 12-150 kDa molecular weight proteins
• Customization—Create your own probes for studying antibody internalization, phagocytosis, and endocytosis
Specifications
Specifications
| For Use With (Application) | Flow Cytometry, Fluoresence Microscopy |
| Product Type | Antibody Labeling Kit |
| Quantity | 3 x 1 mg |
| Sufficient For | 3 x 1 mg Reactions |
| Content And Storage | Store refrigerated. |
| Detection Method | Fluorescence |
| Excitation/Emission | 505/525 nm |
| Labeling Scale | 1 mg |
| Product Line | pHrodo |
| Chemical Reactivity | Amine |
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Frequently Asked Questions (FAQs)
We do not recommend storing antibodies or other proteins labeled with the pHrodo Deep Red Antibody Labeling Kit (Cat. Nos. P35355, P35356) in solutions containing sodium azide, as pHrodo Deep Red is not stable in the presence of sodium azide. Consider using an alternative, non-azide-based anti-microbial.
A DOL determination is required to determine if a suitable amount of dye is attached to the protein. Without a DOL determination, you would not be able to determine if the protein was adequately labeled. A low fluorescent signal may be the same for a protein that is under-labeled or over-labeled, as over-labeling may cause some level of dye-dye quenching.
Yes. The pHrodo Deep Red Antibody Labeling Kit (Cat. Nos. P35355, P35356) was optimized to label either 100 µg or 1 mg of a ~150 kDa protein (molecular weight of a whole IgG) but can be used to label other proteins. Optimize the mass of protein to label based on the molecular weight of the protein relative to the molecular weight of a whole IgG (~150 kDa).