Easily Switch Fluorescence wavelength on your primary antibody
1. Y. Kubota, Y. Oike, S. Satoh, Y. Tabata, Y. Niikura, T. Morisada, M. Akao, T. Urano, Y. Ito, T. Miyamoto, N. Nagai, G. Y. Koh, S. Watanabe and T. Suda, Cooperative Interaction of Angiopoietin-like Proteins 1 and 2 in Zebrafish Vascular Development , Proc. Natl. Acad. Sci. USA, 2005, 102, 13502.
2. T. Yamabuki, A. Takano, S. Hayama, N. Ishikawa, T. Kato, M. Miyamoto, T. Ito, H. Ito, Y. Miyagi, H. Nakayama, M. Fujita, M. Hosokawa, E. Tsuchiya, N. Kohno, S. Kondo, Y. Nakamura, and Y. Daigo, Dikkopf-1 as a Novel Serologic and Prognostic Biomarker for Lung and Esophageal Carcinomas, Cancer Res., 2007, 67, 2517.
3. H. Kohara, Y. Omatsu, T. Suhiyama, M. Noda, N. Fujii and T. Nagasawa, Development of Plasmacytoid Dendritic Cells in Bone Marrow Stromal Cell niches Requires CXCL12-CXCR4 Chemokine Signaling, Blood, 2007, 110, 4153.
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Can I use this kit for other proteins?
Yes, if the molecular weight is greater than 50,000.
Do I have to use a Filtration tube prior to labeling the protein?
If the protein solution does not contain small molecules with amino groups and the concentration of the protein is 10 mg per ml or about 70 μM, there is no need to use the Filtration tube. Just mix 10 μl of the sample solution with 90 μl of Reaction buffer and add the mixture to a vial of NH2-reactive Biotin. After the reaction, transfer all of the reaction mixture to a Filtration tube, and then follow the protocol starting at step 6.
Do I have to use WS buffer to store the biotin-labeled protein?
You don’t have to use WS buffer. You can choose any kind of buffer according to your experiment.
My sample contains small insoluble material. What should I do?
Spin the sample and use the supernatant for the labeling.
How long is the biotin-labeled protein stable?
If you store the biotin-labeled protein at 0-5ºC, it is stable for 2 months. For longer storage, add 100% volume of glycerol, aliquot, and store at -20ºC (if the protein can be frozen). However, please note that stability depends on the protein itself.
What is the minimum amount of IgG that can be labeled by this kit?
The minimum amount is 10 μg IgG; simply follow the protocol. The labeling ratio remains the same for 10 μg to 100 μg of IgG.