A. fumigatus D141, Δasp f3, and asp f3C strains were grown in PD medium at 37 °C for 24 h. The fungal material was separated from culture supernatant by centrifugation at 5,000 g. Proteins were extracted by sonication in presence of protease inhibitor cocktail, and then separated by SDS polyacrylamide electrophoresis (PAGE). 0.5 μg total protein (from ~10 mg dry hyphae) was loaded per lane and recombinant Asp f3 served as a positive control. Protein gels were stained with SimplyBlue SafeStain (Thermo Fisher Scientific) or used for Western blots developed with a previously described rabbit polyclonal anti-Asp f3 antibody and the IRDye 680RD donkey-anti-rabbit antibody (Li-Cor) as the secondary antibody8 (link). The blots were scanned with a Li-Cor Odyssey infra-red Imaging System (Li-Cor Biotechnology, Lincoln, NE).
Irdye 680rd donkey anti rabbit antibody
Manufactured by LI COR
IRDye 680RD donkey-anti-rabbit antibody is a secondary antibody conjugated with the near-infrared fluorescent dye IRDye 680RD. It is designed to detect and quantify rabbit primary antibodies in various immunoassays, such as Western blotting, immunohistochemistry, and ELISA.
Automatically generated - may contain errors
Lab products found in correlation
Simplyblue safestain, by Thermo Fisher Scientific (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Nupage 4 12 bis tris protein gel, by Thermo Fisher Scientific (1 mentions)
Ly294002, by Thermo Fisher Scientific (1 mentions)
Irdye 800cw donkey anti mouse antibody, by LI COR (1 mentions)
Latrunculin b, by Merck Group (1 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Complete edta free protease inhibitor cocktail, by Merck Group (1 mentions)
Sfp synthase, by New England Biolabs (1 mentions)
2 protocols using irdye 680rd donkey anti rabbit antibody
1
Profiling Aspergillus Virulence Factors
2
Insulin Receptor Signaling Pathway Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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CoA-PEG-Nile reds were synthesized as described previously [25] and NR12S was a gift from Dr. Klymchenko [20] . SFP synthase was purchased from NEB. Methyl beta cyclodextrin was purchased from CycloLab. Cholesterol-water soluble, human reconstitute insulin, Complete EDTA-free protease inhibitor cocktails and Latrunculin B were purchased from Sigma-Aldrich. Pierce™ BCA Protein Assay Kit, NuPAGE™ 4-12% Bis-Tris Protein gel and LY294002 was purchased from Thermo Fisher. Phosphatase inhibitor cocktail set V was purchased from Calbiochem. Anti-insulin receptor beta rabbit polyclonal antibody (sc-711) and mouse monoclonal antibody (sc-57342) were purchased from SantaCruz. Anti-Akt mouse monoclonal antibody (Cat. No. 2920), anti-phospho-Akt rabbit monoclonal antibody (Cat. No. 4060) and antiphospho tyrosine rabbit monoclonal antibody (Cat. No. 8954) were purchased from Cell Signaling Tech. Anti-phosphotyrosine mouse monoclonal antibody (PY20) was purchased from BD Transduction Lab. IRDye 800CW donkey antimouse antibody and IRDye 680RD donkey anti-rabbit antibody were purchased from LI-COR biosciences.
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