WBAntibody00001621

Antibody against POS-1, provided by Y. Kohara. pos-1

WBAntibody00003047

Guinea pig anti-HTP-3 1:700 (Y. Kim lab). htp-3

WBAntibody00001856

Antibody against emr-1, from Y. Gruenbaum and K. Wilson. emr-1

WBAntibody00000615

Mouse monoclonal antibody against DPY-7. A DPY-7 fragment constituting most of the predicted mature protein was used to immunize mice, and monoclonal cell lines were generated by fusing splenocytes to Sp2/O-Ag14 myeloma cells by using standard methods. The cell line DPY7-5a was selected for use based on the sensitivity and specificity of the antibody for the DPY-7 protein. It reacts to an epitope within the 40 carboxy-terminal non-Gly-X-Y residues of the protein. dpy-7

WBAntibody00002768

The N-terminal portion (aa 4-77) of CYB-1 was cloned into a maltose-binding protein (MBP)-tagged 6xHis vector, pYS, as follows. First, the MBP gene was PCR-amplified from pHIT198 (a gift from H. Tabara, Tsukuba University, Kyoto, Japan) and cloned into pET- 28a(+) (Novagen, 69864-3) by means of the DNA Ligation Kit Mighty Mix (Takara, 6023) to generate pYS. Second, the N-terminal portion (aa 4-77) of CYB-1 was amplified from yk1306b09 (a gift from Y. Kohara, National Institute of Genetics, Mishima, Japan) and cloned into the pYS vector. The CYB-1-MBP-6xHis fusion protein was expressed and affinity-purified using His-Bind Agarose Resin (ELPIS-BIOTECH, EBE-1031). It was then injected into rats as an antigen to raise an anti-CYB-1 antibody (AbFrontier). This antibody was then affinity-purified from the crude serum, using 6xHis-tagged CYB-1-GST. The GST gene was amplified from pDEST15 (Invitrogen, 11802-014) and inserted into pET-28a(+) (Novagen, 69864-3) to generate pYS1. The same N-terminal portion of CYB-1 used to produce the CYB-1-MBP-6xHis fusion protein was cloned into the pYS1 vector. cyb-1