Primary antibodies used in western blot, immunofluorescence, and immunohistochemistry (IHC) were listed in Supplementary Table 1 . Secondary antibodies include IRDye 800 CW-conjugated anti-rabbit IgG, IRDye 680 CW-conjugated anti-mouse IgG (LI-COR), and horseradish peroxidase (HRP)-conjugated anti-mouse IgG (CST). Secondary antibodies used in immunofluorescence assay included goat anti-rabbit IgG secondary antibody (Alexa Fluor 488) and goat anti-mouse IgG secondary antibody (Alexa Fluor 647) from Invitrogen.
Irdye 680 cw conjugated anti mouse igg
Manufactured by LI COR
The IRDye 680 CW conjugated anti-mouse IgG is a secondary antibody that binds to mouse primary antibodies. This antibody is labeled with the near-infrared dye IRDye 680 CW, which enables detection and quantification of target proteins in Western blotting and other immunoassay applications.
Automatically generated - may contain errors
4 protocols using irdye 680 cw conjugated anti mouse igg
1
Antibody Staining Protocols for WB, IF, and IHC
2
Immunoblotting for Protein Detection
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Western blot was performed as described previously56 (link). Briefly, the whole-cell extract was resolved by SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes. The membranes were blocked in 5% bovine serum albumin and then incubated with diluted primary antibodies overnight at 4°C. Western blot detection was performed with IRDye 800 CW conjugated anti-rabbit IgG or IRDye 680 CW conjugated anti-mouse IgG secondary antibodies according to the manufacturer's protocols (LI-COR Biosciences, Lincoln, NE). The blots were visualized using an Odyssey infrared imaging system (LI-COR Biosciences, Lincoln, NE).
3
Quantitative Western Blot Analysis
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Western blot was performed as described previously55 (link)56 (link). Briefly, the whole-cell extract was resolved by SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes. The membranes were blocked in 5% bovine serum albumin and then incubated with diluted primary antibodies at 4 °C overnight. Western blot detection was performed with IRDye 800 CW conjugated anti-rabbit IgG or IRDye 680 CW conjugated anti-mouse IgG secondary antibodies according to the manufacturer’s protocols (LI-COR Biosciences, Lincoln, NE). The blots were visualized using Odyssey infrared imaging system (LI-COR Biosciences).
4
Western Blot Detection and Visualization
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Western blot was performed as described previously52 (link)53 (link)54 (link). In brief, equal amounts of cell lysates were resolved by SDS-PAGE and then transferred to polyvinylidene fluoride or nitrocellulose membranes. Membranes were blocked and incubated overnight with primary antibodies at 4 °C. The membranes were incubated with appropriate HRP-conjugated secondary antibodies at room temperature for 1 h, and the blots were visualized with PlusECL (KeyGEN BioTECH) according to the manufacturer’s protocol. Alternatively, blots were detected with IRDye 800 CW conjugated anti-rabbit IgG or IRDye 680 CW conjugated anti-mouse IgG secondary antibodies (LI-COR Biosciences, Lincoln, NE), and visualized using Odyssey infrared imaging system (LI-COR Biosciences).
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