Anti IgG (H&L)  

Immunofluorescence of anti rat IgG antibody Tissue: Normal mouse spleen Fixation: user optimized Antigen retrieval: user optimized Primary antibody: rat anti-mouse primary antibody (clone BM8 from eBioscience) against F4/80 (1:100 dilution) Secondary antibody: ATTO dye 488 anti-rat IgG (612-152-120) 1:200 dilution) Localization: macrophage populations throughout the spleen of a normal mouse Staining: antibody as green fluorescent signal with a DAPI blue nuclear counterstain. (Rebecca Evans, The University of Pennsylvania School of Medicine)

Immunofluorescence of anti rat IgG antibody Tissue: Normal mouse spleen Fixation: methanol frozen Antigen retrieval: user optimized Primary antibody: AbD Serotec's Rat anti-mouse CD3 antibody (KT3 clone). (1mg/mll) Secondary antibody: ATTO dye 488 anti-rat IgG (612-152-120) 1:200 dilution) Localization: CD3+ populations throughout the spleen of a normal mouse Staining: antibody as green fluorescent signal with a DAPI blue nuclear counterstain (Rebecca Evans, The University of Pennsylvania School of Medicine)

ATTO R dyes can be used for multicolor immunofluorescent detection with low background and high signal. Examples shown are: A. Tubulin in PtK2- male Rat Kangaroo Kidney Epithelial Cells was detected using ATTO 532 labeled secondary antibody. B. Muscle alpha-actin was stained with a mouse primary antibody and ATTO 488 anti-mouse IgG (green) while Cytokeratin was stained with polyclonal rabbit anti-cytokeratin and ATTO 647N anti-rabbit IgG (red). C. HUVEC (Human umbilical vein endothelial cells were stained with anti- Vimentin-ATTO 532 (green), anti-E-Cadherin-ATTO 655 (red) and DAPI (blue). D. Rat colon sections were stained with Anti-Aquaporin 3-ATTO 594 antibody. Hoechst 33342 (blue) is used as counterstain. Images provided courtesy of Dr. Jorg Reichwein, ATTO-TEC GmbH

Western Blot of Anti-Rat IgG (H&L) (GOAT) Antibody (Min X Bv Ch Gt GP Ham Hs Hu Ms Rb & Sh Serum Proteins) (p/n 612-101-120). 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) (GOAT) Antibody (Min X Bv Ch Gt GP Ham Hs Hu Ms Rb & Sh Serum Proteins) (p/n 612-101-120) 1:1,000 for 60 min at RT. Secondary Antibody: Anti-Goat IgG (DONKEY) Peroxidase Conjugated Antibody (p/n CUST10) 1:40,000 in MB-070 for 30 min at RT. Predicted/Observed 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.