Qdot 705 antibody conjugation kit

To obtain a highly pure monoclonal anti-PAR1 antibody, cloned hybridoma cells producing monoclonal anti-PAR1 antibody were injected into the abdominal cavity of a severe combined immunodeficient (SCID) mouse (Charles River), which is an immunodeficient mouse that lacks immunoglobulins. Ascites were prepared from the mouse, and anti-PAR1 antibody was purified from the ascites using protein G-Sepharose (Amersham Biosciences). The purified antibody was monomerized, mixed with QDs in a molar ratio of approximately 3:1 (antibody: QDs), and then applied to a preparation of anti-PAR1-QDs using a Qdot 705 Antibody Conjugation Kit (Life Technologies), where the number indicates the emission wavelength, as in our previous study28 (link).

We prepared two kinds of quantum dot (QD)‐conjugated antibody complexes for IHC with QDs (IHC‐QDs). One was a QD‐conjugated trastuzumab (Chugai Pharmaceuticals Co. Ltd., Tokyo, Japan) 26, the other was a QD‐conjugated human IgG (Beckman Coulter, Inc., Fullerton, CA). These antibody were monomerized, mixed with QDs in a molar ratio of approximately 3: 1 (antibody: QDs), and then applied to a preparation of antibody‐conjugated QDs using a Qdot 705 Antibody Conjugation Kit (Life Technologies, Waltham, MA), where the number indicates the emission wavelength, as in our previous study.