Sodium dodecyl sulfate-polyacrylamide staining

PCR and in vitro recombination reactions are quite simple and straightforward for generating multiple expression plasmids in parallel, e.g., in a 96-well plate see Fig. 3a). The first preliminary expression experiment was done to evaluate the production level of each GST-fused protein. In this step, we compared the staining patterns of E. coli proteins harboring expression plasmids with the patterns of proteins harboring empty vectors on sodium dodecyle sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under the same culture conditions see Fig. 3b). In addition to... 

Figure 10.7 Polypeptide patterns of material associated with lipid globules of cow s milk. The sodium dodecyl sulfate-polyacrylamide gel in (a) was stained with coomassie blue, and polypeptides in the gel in (b) were visualized with the more sensitive silver stain (Merril ef at. 1981). In both (a) and (b) the left lane contains proteins extracted directly from washed milk...

In Experiment 4, your sample of a-lactalbumin extracted from bovine milk was subjected along with other proteins to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). After staining with the dye Coomassie Blue, deeply colored bands appeared on the gel wherever there was a protein. You suspected that some of the blue bands on the gel were due to a-lactalbumin. If molecular weight standards were included on the slab gel, you were able to estimate the molecular weight for a-lactalbumin and other proteins. SDS-PAGE is indeed a very effective analytical tool to achieve fractionation of protein mixtures, to analyze purity, and to estimate molecular weight, but it provides no experimental data to prove the identity... 

In general, a crude or partially purified extract is electrophoresed on a sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE) then the protein band is lightly stained and cut out. In the simplest method, the acrylamide gel band is reduced to a pulp, mixed with Freund s adjuvant, and injected. Unfortunately, this technique is not always successful. Its failure can probably be attributed to factors such as the difficulty of disaggregating the acrylamide, the difficulty with which the protein diffuses from the gel, the presence of SDS in large quantities resulting in extensive tissue and cell damage, and finally, the toxicity of the acrylamide... 

Femandez-Patron, C., Castellanos-Serra, L., and Rodriguez, P. (1992) Reverse staining of sodium dodecyl sulfate polyacrylamide gels by imidazole-zinc salts sensitive detection of unmodified proteins. Biotechniques 12, 564-573. 

U Tessmer, R Dernick. Preparative separation of poliovirus structural polypeptides by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, copper staining and electroelution, and induction of monospecific antisera. Electrophoresis 10 277-279, 1989. 

Figure 10. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of products of limited proteolysis of the debranching enzyme with trypsin. The molecular weights shown of the various bands were determined by the methodology described previously (26). The ratio of debrancher to trypsin was 100 to 1. The incubation was conducted for 60 minutes at 25°. The gel stain was Coomassie Brilliant Blue and the absorbance was measured at 600 nm using a Gilford gel scanner with a 0-1 O.D. chart scale (36).

Kido, N., Ohta, M., Kato, N. Detection of lipopolysaccharides by ethidium bromide staining after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. J Bacteriol 172 (1990) 1145-1147. 

Figure 6-4. Sodium dodecyl sulfate polyacrylamide gel electrophoresis pattern of normal red cell membrane proteins with Coomassie blue staining.

Salinovich, O. and Montelaro, R. C. (1986) Reversible staining and pepude mapping of proteins transferred to nitrocellulose after separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. AnaL Biochem 156, 341-347. 

Schematic representations of sodium dodecyl sulfate polyacrylamide gel electrophoretic patterns of red blood cell membranes (M) and membrane skeletons (S), based on work by Fairbanks and Steck. Proteins are stained with Coomassie blue (CB) and sialoglycoproteins with periodic acid-Schiff (PAS). GPA, GPB, and GPC are glycophorin A. B, and C, respectively G3PD is glyceraldehyde-3-phosphate dehydrogenase. (GPA)2 and (GPB)2 are dimers, and GPA-GPB is a heterodiraer. [Reproduced with permission from J. B. Stanbury, J. B. Wyngaarden, D. S. Fredrickson, et al. (Eds.), The Metabolic Basis of Inherited Disease, 5th ed. McGraw-Hill, New York, 1983.]...

FIGURE 5 Immunoprecipitation of vimentin by anti-protein kinase G (PKG) from rat aortic smooth muscle cells (SMCs). PKG was immunoprecipitated from low-passaged (passage 1) SMCs the proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose. Vimentin was visualized using mouse antivimentin in a Western blot. The antibody also recognizes isotopes of rabbit anti-bovine PKG, as indicated by the staining of the immunoglobulin G heavy chain. [Data from MacMillan-Crow and Lincoln (1994) with permission of the American Chemical Society.]... 

Polyacrylamide Gel Electrophoresis and Fluorography. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was carried out using a 10-17% linear polyacrylamide gradient slab gel with a 1 cm 5% stacking gel that incorporated the buffer system of Laemmli ( ). Samples were prepared for electrophoresis and gels were run, fixed, and stained for protein as previously described (23). 

Fig. 2. The representative results for checking quality of recombinant proteins checked by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gel. Two microgram of SH2 recombinant proteins were loaded on 10% SDS-PAGE gel, and the gels were stained with Coomassie Brilliant Blue.

Myers, J., et al. (1996). A Method for Enhancing the Sensitivity and Stability of Stains-all for Phosphoproteins Separated in Sodium Dodecyl Sulfate-polyacrylamide Gels, Anal. Biochem. 240 300 302. 

The fixing, staining, and destaining of sodium dodecyl sulfate-polyacrylamide (SDS-PAGE) gels and polyvinylidene-difluoride (PVDF) membranes was one of the first... 

Figure 2. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and silver staining of active fractions from each chromatographic stage of the purification of Drosophila heat shock activator protein. The lanes indicate active chromatographic fractions from 1. heparin-Sepharose. 2. specific DNA-Sepharose (second column the most enriched fraction from the first column shows a slightly more complex protein gel pattern). 3. FPLC Mono S.

Note The elution should be optimized by checking at least two to three different elution solvents. Eluates may be assessed using a one-dimensional (ID) sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel stained with colloidal Coomassie. 

Lin, C. L. Chen, H. J. Hou, W. C. Activity staining of glutathione peroxidase after electrophoresis on native and sodium dodecyl sulfate polyacrylamide gels. Electrophoresis 2002, 23, 513-516. 

---