Supernatants of the hybridomas Hermes-1 (9B5, anti-human CD44, used as an isotype-matched control) and M1/9 (anti-mouse CD45) were produced in our own laboratory and used undiluted. Mec13.3 (anti-mouse PECAM1) was a gift of Dr E. Dejana (IFOM, Milan, Italy). A polyclonal rabbit anti-mouse DARC antibody against the C-terminus (amino acids 319–333: LPRQASQMDALAGK) as previously published (Kashiwazaki et al., 2003 (link)) was prepared by Sigma-Genosys, Sigma Aldrich. Purified IgG fractions of the antisera were obtained by protein A affinity chromatography. Specific recognition of DARC by the polyclonal IgG fraction was confirmed by positive immmunoreactivity on spotted peptides, western blots of lymph nodes and specific staining of high endothelial venules of peripheral lymph nodes in frozen tissue sections of wild-type mice and absence of this staining in Darc−/− mice. Secondary antibodies used were Alexa Fluor® 488 goat-anti rat IgG, Cy3 goat anti rat IgG, biotin goat anti-rat IgG combined with AMCA-streptavidin and Cy3-goat anti rabbit IgG (all from Jackson ImmunoResearch Laboratories).
Amca streptavidin
Manufactured by Jackson ImmunoResearch
Sourced in United States
AMCA-streptavidin is a fluorescent conjugate of the protein streptavidin. Streptavidin binds strongly to the small molecule biotin, which is commonly used to label biological targets. The AMCA fluorescent dye is attached to the streptavidin, allowing it to be detected and visualized when bound to biotinylated samples.
Automatically generated - may contain errors
Lab products found in correlation
Cy3 goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Cy3 goat anti rat igg, by Jackson ImmunoResearch (1 mentions)
Sigma genosys, by Merck Group (1 mentions)
Alexa fluor 488 goat anti rat igg, by Jackson ImmunoResearch (1 mentions)
Rotenone, by MP Biomedicals (1 mentions)
T0198, by Merck Group (1 mentions)
2 protocols using amca streptavidin
1
Immunodetection of Mouse DARC Protein
2
Antibodies and Reagents for Studying Alpha-Synuclein
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Reagents and antibodies were obtained from the following sources: anti-αSyn antibody (1:1000; 4B12, GeneTex, Irvine, CA, USA); anti-FABP5 antibody (1:1000; AF3077, R&D Systems, Minneapolis, MN, USA); anti-β-tubulin antibody (1:3000; T0198, Sigma-Aldrich, St Louis, MO, USA); anti-VDAC antibody (1:1000; 4866S, Cell Signaling Technology, Danvers, MA, USA); anti-rabbit or mouse IgG antibody conjugated with horseradish peroxidase (1:5000; Southern Biotech, Birmingham, AL, USA); anti-goat IgG antibody conjugated with peroxidase (1:5000; Rockland Immunochemical, Limerick, PA, USA); Alexa 594-labeled anti-mouse IgG, Alexa 488-labeled anti-Goat IgG and Alexa 594-labeled anti-rabbit IgG antibody (1:500; Invitrogen, Waltham, MA, USA); Biotin-SP anti-Goat IgG antibody (1:500; Jackson ImmunoResearch, West Grove, PA, USA); and AMCA streptavidin (1:500; Jackson ImmunoResearch). These materials were used according to the manufacturers’ datasheets. Rotenone was purchased from MP Biomedicals (150154; Santa Ana, CA, USA). All other reagents were obtained from FUJIFILM Wako Pure Chemicals (Osaka, Japan) unless otherwise noted. The FABP5 ligand (ligand 7) was described in a previous report [24 (link)].
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