Anti IgG (H&L)   DyLight (商標名)

DyLight^TM dyes can be used for two-color western blot detection with low background and high signal. Anti-tubulin was detected using a DyLight^TM 680 conjugate. Anti-TNFa was detected using a DyLight^TM 800 conjugate. The image was captured using the Odyssey^(R) Infrared Imaging System developed by LI-COR.

Western Blot of Anti-Rat IgG (H&L) (RABBIT) Antibody (Min X Human Serum Proteins) (p/n 612-4126). Lane M: 3 μl Molecular Ladder. Lane 1: Rat IgG whole molecule (p/n 012-0102). Lane 2: Rat IgG F(c) Fragment (p/n 012-0103). Lane 3: Rat IgG Fab Fragment (p/n 012-0105). Lane 4: Rat IgM Whole Molecule (p/n 012-0107). Lane 5: Rat Serum (p/n D310-05). All samples were reduced. Load: 50 ng per lane. Block: MB-070 for 30 min at RT. Primary Antibody: Anti-Rat IgG (H&L) (RABBIT) Antibody (Min X Human Serum Proteins) (p/n 612-4126) 1:1,000 for 60 min at RT. Secondary Antibody: Anti-Rabbit IgG (GOAT) Peroxidase Conjugated Antibody (p/n 611-103-122) 1:40,000 in MB-070 for 30 min at RT. Predicted/Obsevered Size: 25 and 55 kDa for Rat IgG and Serum, 25 kDa for F(c) and Fab, 78 and 25 kDa for IgM. Rat F(c) migrates slightly higher.

Dot Blot showing the detection of Rat IgG. A three-fold serial dilution of Rat IgG starting at 200ng was spotted onto 0.45 ?m nitrocellulose. After blocking in Blocking Buffer for Fluorescent Western Blotting (p/n MB-070) 1 Hour at 20C, Anti-Rat IgG (H&L) (RABBIT) Antibody DyLight^TM 800 Conjugated (Min X Human Serum Proteins) (p/n 612-445-026) secondary antibody was used at 1:5000 in Blocking Buffer for Fluorescent Western Blotting (p/n MB-070) and imaged on the LiCor Odyssey imaging system.

Properties of DyLight^TM Conjugates.