Figure 2: Anti-RAINBOW antibody detects multi-colour Remazol prestained molecular weight markers. | Scientific Reports

Figure 2: Anti-RAINBOW antibody detects multi-colour Remazol prestained molecular weight markers.

From: Anti-RAINBOW dye-specific antibodies as universal tools for the visualization of prestained protein molecular weight markers in Western blot analysis

Figure 2

(a) Presence of antibodies against four Remazol dyes in mouse sera. Immune sera of mice (3rd bleed) immunized with different prestained proteins were tested with a Miniblotter 28 channel unit against Remazol dye stained phosphorylase b (orange), γ-globulin heavy chain (red), ADH (blue), and γ-globulin light chain (yellow). M1, M2, M3, M4: mouse 1, 2, 3, 4. 20× more dye stained proteins were loaded on the colorimetric marker lane than on the lanes used for Western blot analysis with the mouse sera. (b) The anti-RAINBOW monoclonal antibody shows equal detection of Remazol-dye stained proteins: phosphorylase b (red), BSA and ADH (blue), chymotrypsin (yellow) and lysozyme (orange). 10× more dye stained proteins were loaded on the colorimetric lane shown in the left panel than on the lane incubated with anti-RAINBOW antibody. (c) anti-RAINBOW monoclonal antibody detects all marker bands in a variety of commercial multicolor protein marker mixtures. CST: Cell Signaling Technology, Color-coded Prestained Protein Marker, High Range (12949P); NEB: New England Biolabs, ColorPlus Prestained Protein Marker (P7709); Thermo: Thermo Scientific: PageRuler Plus Prestained Protein Ladder (26619). 5× more of each protein marker mixture was loaded on the colorimetric lanes with the visible marker bands than on the lanes incubated with anti-RAINBOW antibody. (d) anti-RAINBOW monoclonal antibody does not cross-react with unstained cellular proteins. 1: E. coli; 2: A. thaliana; 3: S. cerevisiae; 4: C. elegans; 5: D. melanogaster; 6: Chicken follicle; 7: CHO, Chinese hamster ovary; 8: Rat1, rat fibroblasts; 9: N2a, mouse neuroblastoma; 10: CV1, Green monkey epithelial; 11: Hela, human epithelial; M: homemade prestained rainbow marker (RF3B-phosphorylase b, RBBR-BSA, RO16-ADH, RGY-γ-globulin light chain). (e) anti-RAINBOW monoclonal antibody cross-reacts with Remazol dyes not used for immunization. Lane 1: RBBR-ADH (blue), lane 2: RF3B-phosphorylase b (red), lane 3: RO16-ADH (orange), lane 4: RGY-γ-globulin light chain (yellow), lane 5: RB5-phosphorylase b (black), lane 6: RV5R-γ-globulin heavy chain (violet). 5× more Bio-Rad marker proteins and 10× more of the Remazol stained proteins were loaded on the upper blot (colorimetric image) than on the lower blot incubated with anti-RAINBOW antibody.

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