Frequently Asked Questions

1. Why use ChromaLink Biotin or ChromaLink Digoxigenin labeling kits?
2. Can all antibodies and/or other proteins be labeled using ChromaLink reagents?
3. What’s the difference between All-in-One kits and other antibody conjugation kits?
4. Is the specific activity of the enzymes used in our conjugation kits (AP/HRP) high?
5. What if I want to label more antibody or protein than the kit allows?
6. My antibody sample contains BSA, gelatin, glycine, or Tris. Is that ok?
7. How long is the conjugate stable?  How should I store my conjugate?
8. I have used other kits in the past, why are these kits better?
9. What are the advantages of using high biotin binding capacity bead?
10. Do I need to block my beads?  How?
11. My beads arrived clumped, what should I do?
12. Can I heat the beads to cleave the biotin/streptavidin bond?
13. Do you have beads for direct binding of antibodies, peptides or oligos?
14. How do I know which linker to put on my 2 different biomolecules?
15. What are the advantages of using a quantifiable linker?
16. How long are the linkers stable on my modified biomolecules?

Antibody Labeling Kits (ChromaLink Biotin, ChromaLink DIG, HRP AP and R-PE All-in-One Kits)

Why use ChromaLink Biotin or ChromaLink Digoxigenin labeling kits?

For maximum sensitivity and reproducibility it is important to avoid over labeling, which leads to reduced immuno reactivity, or underlabeling, which leads to weak signal. ChromaLink Biotin and Digoxigenin Labeling Kits are the only kits that allow for tracing and quantification of the incorporated label with a simple UV scan of the labeled protein. By simply measuring the Biotin or DIG labeled protein at 280 and 354nm and then inserting those values into an easy to use calculator, you can determine the exact number of biotins or DIG on your antibody.

Can all antibodies and/or other proteins be labeled using ChromaLink reagents?

Yes, all proteins and antibodies may be labeled with the ChromaLink Biotin or DIG labels. These products label primary amino groups on lysines found in proteins; a cysteine labeling ChromaLink Biotin is also available.

What’s the difference between All-in-One kits and other antibody conjugation kits?

These are the only kits that allow you to isolate virtually 100% pure antibody conjugate without HPLC/FPLC column purification. The All-in-One Kits used for antibody labeling are the only products of their kind that convert 100% of antibody to conjugate, making purification easy with the Solulink mini-purification spin columns. All-in-One kits are the only kits for making high yielding, pure, high-quality antibody conjugate.

Is the specific activity of the enzymes used in our conjugation kits (AP/HRP) high?

Yes, we use the highest quality enzymes so that your conjugates produce good results. The activity of the HRP used in our labeling kits is >300 purpuroglain U/mg and the Alkaline Phosphatase is >7000 DEA U/mg. These highly active enzymes ensure your conjugates perform to your expectations.

What if I want to label more antibody or protein than the kit allows?

We can work with you to provide all of the necessary components to scale up your reactions. Our custom service department can also make your scaled up conjugates for you.

My antibody sample contains BSA, gelatin, glycine, or Tris. Is that ok?

Large carrier proteins such as BSA or gelatin must be removed by Protein A or G columns. Small molecule contaminants such as glycine and Tris are removed with the included desalting columns.

How long is the conjugate stable?  How should I store my conjugate?

The conjugate bond is stable indefinitely. The conjugate storage conditions are indicated in the user manuals for each kit.

I have used other kits in the past, why are these kits better?

Each of Solulink’s products are designed to provide the user with several specific advantages;

• ChromaLink Labeling Kits- Controlled and reproducible labeling results using innovative traceable linkers.
• All-in-One Kits- Easy-to-use kits with purification included, > 60% yields.
• Conjugation Kits and Reagents- Include high yields, fast catalyzed reaction times, controlled and highly efficient conjugation (100% conjugate formation), stable bonds and traceable quantitative linkers.

Other conjugation companies still use old linkers, with all their inherent shortcomings, to label and/or conjugate biomolecules. Solulink is the only company to use new and improved methods for next generation linkers and kits to outperform the competition.

Beads

What are the advantages of using high biotin binding capacity bead?

Higher binding capacity means greater capture with less bead mass. This requires less bead mass for your immobilizations assays resulting in better assay capture and in overall savings.

Do I need to block my beads?  How?

Depending on your application, Solulink recommends blocking Streptavidin beads. If you are capturing non-phosphorylated proteins we recommend blocking with Bead Blocking Solution. If you are capturing DNA or RNA, we recommend blocking with yeast tRNA, salmon sperm or herring sperm DNA at 10mg.mL, if the presence of blocking DNA or RNA are not a factor in your down stream application.

My beads arrived clumped, what should I do?

It is normal for the beads to have mild aggregation after shipping. It is important to re-suspend the beads with light sonication or vortexing. Please follow the General Procedure for Ultrasonication in the Bead Blocking instruction manual.

Can I heat the beads to cleave the biotin/streptavidin bond?

As with all streptavidin beads, excessive heating will lead to dissociation of the streptavidin tetramer and leaching of monomeric subunits. We don’t recommend heating the Streptavidin beads over 40 ᵒC.

Do you have beads for direct binding of antibodies, peptides or oligos?

The NanoLink 4FB Beads are used for direct binding of modified biomolecules. To learn more please visit the NanoLink 4FB product page.

Linkers- for covalent crosslinking

How do I know which linker to put on my 2 different biomolecules?

Please check our product selection guide for a detailed explanation. If you would like recommendations, we welcome you to contact customer support for a free consultation.

What are the advantages of using a quantifiable linker?

With the Solulink linkers, you can easily quantitate the number of linkers you add to each biomolecule. As with labeling, for maximum sensitivity and reproducibility it is important to avoid too many linkers, which leads to reduced immune reactivity, or too few linkers, which leads to weak signal or unconjugated material. Quantitative linkers allow for more control and reproducible steps during the conjugation reaction.

How long are the linkers stable on my modified biomolecules?

4FB linkers are extremely stable on the activated biomolecule and have remained 100% active after over a year at 4ᵒC. The HyNic modified biomolecules should be reacted to the 4FB counterpart within 3 hours of modification.