ICG Labeling Kit – NH2


– Suitable wavelength for In Vivo imaging
– Quick and Easy Labeling to Antibodies : 1.5hr
– High Recovery Rate : more than 90%

Contents of the Kit:

Storage: 0-5ºC
Shipping Condition: ambient temperature

DescriptionReferencesDataQ & AManualS.D.S

Product Description
ICG Labeling Kit – NH2 is used primarily for the preparation of ICG (Iindocyanin green)-labeled antibody for near-infrared fluorescence imaging. ICG offers two remarkable properties:

1) ICG has a strong near-infrared fluorescence even after a few days under physiological conditions. The excitation and emission wavelength of the ICG-labeled proteins are 774 nm and 805 nm, respectively.
2) ICG has been used in clinical fields such as a hepatic deficiency testing. Therefore, ICG and ICG conjugates are materials suitable for in vivo imaging.

This kit contains all required compornents required for labeling, including storage buffer for conjugates. The labeling process is simple:. Add NH2-reactive ICG to protein solution on a filter membrane, and incubate at 37ºC, for 10 minutes. A filtration tube can remove excess ICG molecules.

Fig. 1 Fluorescein Labeling Process to IgG

♦ If the IgG solution contains other proteins with molecular weight greater than 10,000, such as serum albumin or gelatin, purify the IgG solution before labeling fluorescein with this kit. Commercially available antibody may contain BSA or gelatin as a stabilizer. Dojindo offers IgG Purification Kit-A (AP01-10) and IgG Purification Kit-G (AP02-10) for the purification of the IgG solution

1) M. Ogawa, C. A. S. Regino, J. Seidel, M. V. Green, W. Xi, M. Williams, N. Kosaka, P. L. Choyke and H. Kobayashi, “Dual-Modality Molecular Imaging Using Antibodies Labeled with Activatable Fluorescence and a Radionuclide for Specific and Quantitative Targeted Cancer Detection”Bioconjugate Chem., 2009, 20(11), 2177.
2) M. Ogawa, N. Kosaka, P. L. Choyke and H. Kobayashi, “In vivo Molecular Imaging of Cancer with a Quenching Near-Infrared Fluorescent Probe Using Conjuates of Monoclonal Antibodies and Indocyanine Green”Cancer Res., 2009, 69(4), 1268.
3) N. Kosaka, M. Ogawa, P. L. Choyke and H. Kobayashi, “Clinical implications of near-infrared fluorescence imaging in cancer”Future Oncology2009, 5(9), 1501.
4) S. Ito, N.Muguruma, S. Hayashi, S. Taoka, T. Bando, K. Inayama, M. Sogabe, T. Okahisa, S. Okamura, H. Shibata, T. Irimura, K. Takesako and S. Shibamura, “Development of Agents for Reinforcement of Fluorescence on Near-infrared Ray Excitation for Immunohistological Staining”Bioorg. Med. Chem., 1998, 6, 613.
5) S. Ito, N. Muguruma, Y. Kakehashi, S. Hayashi, S. Okamura, H. Shibata, T. Okahisa, M. Kanamori, S. Shibamura, K. Takesako, M. Nozawa, K. Ishida and M. Shiga, “Development of Fluorescence-Emitting Antibody Labeling Substance by Near-Infrared Ray Excitation”Bioorg. Med. Chem. Lett., 1995, 5, 2689.
6) K. Inayama, S. Ito, N. Muguruma, Y. Kusaka, T. Bando, Y. Tadatsu, M. Tadatsu, K. Ii, S. Shibamura and K. Takesako, “Basic Study of an Agent for Reinforcement of Near-infrared Fluorescence on Tumor Tissue”Digestive and Liver Disease2003, 35, 88.
7) S. Ito, N. Muguruma, S. Hayashi, S. Taoka, T. Bando, Y. Kusaka, M. Yano, S. Ichikawa, A. Hiasa, T. Omoya, H. Honda, I. Shimizu, K. Ii, K. Nakamura, K. Takesako, Y. Goto and S. Shibamura, “Visualization of Human Gastric Cancer with a Novel Infrared Fluorescent Labeling Marker of Anti-carcinoembryonic Antigen Antibody in vitro”Dig. Endosc., 2000, 12, 33.
8) S. Taoka, S. Ito, N. Muguruma, S. Hayashi, Y. Kusaka, K. Ii, K. Nakamura, K. Imaizumi, K. Takesako and S. Shibamura, “Reflected Illumination-type Imaging System for the Development of Infrared Fluorescence Endoscopy”Dig. Endosc.,1999, 11(4), 321.
9) S. Ito, N. Muguruma, S. Hayashi, S. Taoka, A. Tsutsui, T. Fukuda, T. Okahisa, Y. Ohkita, H. Matsunaga, I. Shimizu, K. Nakamura, K. Imaizumi, K. Takesako and S. Shibamura,“Development of an Imaging System Using Fluorescent Labeling Substances Excited by Infrared Rays”Dig. Endosc., 1997, 9, 278.
10) S. Ito, N. Muguruma, Y. Kusaka, M. Tadatsu, K. Inayama, Y. Musashi, M. Yano, T. Bando, H. Honda, I. Shimizu, K. Ii, K. Takesako, H. Takeuchi and S. Shibamura, “Detection of Human Ganstric Cancer of Resected Specimens Using a Novel Infrared Fluorescent Anti-Human Carcinoembryonic Antigen Antibody with an Infrared Fluorescence Endoscope in Vitro”Endoscopy2001, 33(10), 849.
11) N. Muguruma, S. Ito, T. Bando, S. Taoka, Y. Kusaka, S. Hayashi, S. Ichikawa, Y. Matsunaga, Y. Tada, S. Okamura, K. Ii, K. Imaizumi, K. Nakamura, K. Takesako and S. Shibamura, “Labeled Carcinoembryonic Antigen Antibodies Excitable by Infrared Rays: a Novel Diagnostic Method for Micro Cancers in the Digestive Tract”Internal Medicine1999, 38(7), 537.
12) T. Bando, N. Muguruma, S. Ito, Y. Musashi, K. Inayama, Y. Kusaka, M. Tadatsu, K. Ii, T. Irimura, S. Shibamura and K. Takesako, “Basic Study on a Labeled anti-mucin Antibody Detectable by Infrared-fluorescence Endoscopy”J. Gastroenterol., 2002, 37, 260.
13) N. Muguruma, S. Ito, S. Hayashi, S. Taoka, H. Kakehashi, K. Ii, S. Shibamura and K. Takesako, “Antibodies Labeled with Fluorescence-agent Excitable by Infrared Rays”J. Gastroenterol., 1998, 33, 467.
14) W. Aung, A. Tsuji, H. Sudo, A. Sugyo, T. Furukawa, Y. Ukai, Y. Kurosawa and T. Saga, “Immunotargeting of Integrin α6β4 for Single-Photon Emission Computed Tomography and Near-Infrared Fluorescence Imaging in a Pancreatic Cancer Model”Molecular Imaging2016, 15, 1.

Fig. 3 Fluorescent Property of ICG Dye

Can I use this kit for F(ab')2?

Yes, please follow the labeling protocol for IgG. The recovery of the conjugate should be over 80%.

Can I use this kit for other proteins or peptides?

Yes, if the molecular weight is greater than 50,000.

Can I use this kit to label oligopeptides or oligonucleotides?

No. Oligonucleotides and oligopeptides may be too small to retain on the membrane filter of Filtration Tube.

What is the minimum amount of IgG that can be labeled with LK31-10?

The minimum amount is 50 μg. There is no significant difference in sensitivity and background between 50 μg and 200 μg of IgG. Though 10 μg IgG can still be labeled using this kit, the background will be higher.

How many ICG molecules are introduced to protein?

The number of conjugated ICG depends on the protein. In the case of rabbit IgG, one molecule of ICG conjugate to each protein molecule.

Do I have to use a Filtration tube prior to labeling the protein?

If the protein solution does not contain small molecules with an amino group and the concentration of the protein is 10 mg per ml, or about 70 μM, there is no need to use the Filtration tube. Mix 10 μl of the sample solution with 90 μl of Reaction buffer and add 8 μl NH2-reactive ICG (prepared at step 3) to the mixture, and follow the protocol starting at step 4.

Do I have to use Storage buffer included with the kit?

No, you don’t have to use Storage buffer from the kit. You can choose any kind of buffer appropriate for your experiment.

My sample contains small insoluble material. What should I do?

Spin the sample and use the supernatant for labeling.

Related Categories
Labeling Chemistry

Related Products

COVID-19 Announcement

Due to rising concerns regarding the COVID-19 (Coronavirus 2019) outbreak, Dojindo Molecular Technologies, Inc will be taking the necessary steps to prioritize public health and safety.

Scroll to Top