The Reliability of Molecular Weight Determinations by Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis

Weber, Klaus; Osborn, Mary

doi:10.1016/s0021-9258(18)94333-4

Cited by 19,639 publications

(1,427 citation statements)

References 44 publications

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“…The gels were then incubated for 60 min in this solution at 37°C and then washed and fixed as previously described. According to Weber & Osborn (1969), the staining procedure was performed using a 0.1% coomassie brilliant blue R-250 solution, while distaining was carried out in a 35:10:55 methanolacetic acid-water solution.…”

Section: Electrophoretic Analysesmentioning

confidence: 99%

Partial characterization of digestive proteases in Pacific red snapper Lutjanus peru Nichols & Murphy, 1922 (Perciformes: Lutjanidae)

Peña-Marín

Ibarra‐Castro

Martínez-Brown

et al. 2021

Lat. Am. J. Aquat. Res.

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Pacific red snapper (Lutjanus peru) is an important commercial species in Mexico with great aquaculture potential; however, digestive physiology is still unknown. Therefore, the objective of the present work was to characterize the digestive proteases of L. peru juvenile using biochemical and electrophoretic techniques. Results showed a higher acid protease activity than the alkaline proteases, trypsin, chymotrypsin, and leucine aminopeptidase (LAP). The optimum temperature for acid proteases was between 30 to 40°C. Trypsin activity showed two maximum peaks of temperature (30 and 50°C), while alkaline proteases, chymotrypsin, and LAP had optimum temperatures of 50, 50 to 60, and 40°C, respectively. Moreover, the optimum pH of acid proteases was between 2 and 3. Also, alkaline proteases, trypsin, chymotrypsin showed pH optimums at pH 6, 9, and 5, respectively, although LAP showed two optimum pH values at 6 and 9. Acid protease zymogram showed three isoforms, totally inhibited by pepstatin A. Alkaline protease zymogram revealed six bands (125.4, 67.2, 57.9, 48.6, 29.8, and 26.9 kDa), which were inhibited by specific serine-proteases and metalloproteases inhibitors. In conclusion, the main digestion in L. peru depends on stomach proteases, which are characteristic of carnivorous fish, followed by intestinal digestion supported mainly by chymotrypsin.

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Section: Electrophoretic Analysesmentioning

confidence: 99%

Partial characterization of digestive proteases in Pacific red snapper Lutjanus peru Nichols & Murphy, 1922 (Perciformes: Lutjanidae)

Peña-Marín

Ibarra‐Castro

Martínez-Brown

et al. 2021

Lat. Am. J. Aquat. Res.

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“…Precipitated proteins were separated by 5% SDS-PAGE at pH 6.0 according to Weber and Osborn 44 . The radioactivity associated with the separated Ca 2+ -ATPase was quantified by digital autoradiography as described 45 .…”

Section: Methodsmentioning

confidence: 99%

Angle change of the A-domain in a single SERCA1a molecule detected by defocused orientation imaging

Katoh

Daiho

Yamasaki

et al. 2021

Sci Rep

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The sarcoendoplasmic reticulum Ca2+-ATPase (SERCA) transports Ca2+ ions across the membrane coupled with ATP hydrolysis. Crystal structures of ligand-stabilized molecules indicate that the movement of actuator (A) domain plays a crucial role in Ca2+ translocation. However, the actual structural movements during the transitions between intermediates remain uncertain, in particular, the structure of E2PCa2 has not been solved. Here, the angle of the A-domain was measured by defocused orientation imaging using isotropic total internal reflection fluorescence microscopy. A single SERCA1a molecule, labeled with fluorophore ReAsH on the A-domain in fixed orientation, was embedded in a nanodisc, and stabilized on Ni–NTA glass. Activation with ATP and Ca2+ caused angle changes of the fluorophore and therefore the A-domain, motions lost by inhibitor, thapsigargin. Our high-speed set-up captured the motion during EP isomerization, and suggests that the A-domain rapidly rotates back and forth from an E1PCa2 position to a position close to the E2P state. This is the first report of the detection in the movement of the A-domain as an angle change. Our method provides a powerful tool to investigate the conformational change of a membrane protein in real-time.

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“…Electrophoretic analysis of the purified phycocyanin was carried out on 12% non-denaturing SDS-PAGE [19], while its molecular mass was detected [20]. The proteins were visualized by Silver stain [21].…”

Section: Electrophoretic Analysismentioning

confidence: 99%

C-Phycocyanin isolated from Microcystis aeruginosa Kützing mitigates renal injury induced by potassium dichromate via toll-like receptor-4 down regulation in rats

et al. 2021

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C-Phycocyanin is among the most promising microalgal derived molecules with an array of pharmacological activities. Toll-like receptors (TLRs) have a vital role in regulating inflammatory response in renal injury. The aim of this study was to evaluate the possible renal therapeutic effect of phycocyanin isolated from Microcystis aeruginosa Kützing harvested from high-rate algal pond on potassium dichromate (PD)-induced renal injury in rats. Rats were assigned randomly into: Group I: Normal control group. Group II: Rats injected with a single injection of PD (15 mg/kg; s.c) and served as renal injury group. Group III and IV: Rats received C-phycocyanin (25 & 50 mg/kg body weight; orally), after PD injection, daily for 7 days. Injection of PD induced a marked renal injury, evidenced by a significant increase in kidney functions and renal contents of malondialdehyde (MDA), tumor necrosis factor-alpha (TNF-α), toll-like receptor 4 (TLR4), heat shock protein 70 (HSP70) and insulin growth factor-1 (IGF-1) In addition PD reduced renal content of reduced glutathione (GSH) and exhibited tubular degeneration and necrosis. C-phycocyanin has renal therapeutic effect against renal injury induced by PD through improving kidney function, down regulation of oxidative stress and TLR4/TNF-α/HSP70 inflammatory pathway as well as modulation of IGF-1.

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The Reliability of Molecular Weight Determinations by Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis

Cited by 19,639 publications

References 44 publications

Partial characterization of digestive proteases in Pacific red snapper Lutjanus peru Nichols & Murphy, 1922 (Perciformes: Lutjanidae)

Partial characterization of digestive proteases in Pacific red snapper Lutjanus peru Nichols & Murphy, 1922 (Perciformes: Lutjanidae)

Angle change of the A-domain in a single SERCA1a molecule detected by defocused orientation imaging

C-Phycocyanin isolated from Microcystis aeruginosa Kützing mitigates renal injury induced by potassium dichromate via toll-like receptor-4 down regulation in rats

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