Seeing gel wells well

Smith, Ivor; Cromie, Ruth; Stainsby, Karen

doi:10.1016/0003-2697(88)90297-7

Cited by 17 publications

(4 citation statements)

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“…Electrophoresis was carried out at 100 V for 2.5 hours. The gels were stained with Coomassie Brilliant Blue R-250 (Smith et al 1988) and visualised and/or transferred to a Polyvinylidene Fluoride (PVDF) membrane (450 nm, USA) for Western blotting. Immunoblotting was carried out under a constant current of 0.8 mA per square cm of the membrane for 3 h, following Towbin and Gordon (1984) with slight modifications.…”

Section: Methodsmentioning

confidence: 99%

Optimisation of an extraction technique of fish allergens suitable for detection and diagnosis

Ma¹,

Ramesh²,

Li³

et al. 2017

Czech J. Food Sci.

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Ma J., Pavase T., Li Z., Lin H. (2017): Optimisation of an extraction technique of fish allergens suitable for detection and diagnosis. Czech J. Food Sci., 35: 24-31.An optimised protocol for the extraction of allergenic proteins in fish was developed. Twelve existing or modified extraction buffers were evaluated based on the amount and quality of obtained 12 kDa IgG-binding protein (major fish allergen parvalbumin) by SDS-PAGE, immunoblotting, and ELISA. The results indicated that using DL-Dithiothreitol during the extraction process, the stability and functionality of the final extract were significantly increased. Solution 3 (containing Tris-HCl, Glycine, and DL-Dithiothreitol) yielded the highest amount of parvalbumin. The extract also induced a higher allergenic reactivity to the three tested human sera by immunoblotting. The results indicated that solution 3 provided better results in fish allergen detection and diagnosis. In conclusion, our study discovered and established solution 3 (Tris-HCl + Glycine + DTT) as the most potential buffer to overcome the conventional allergen extraction buffer and therefore it is highly recommended as a substitute optimised extraction buffer.

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Section: Methodsmentioning

confidence: 99%

Optimisation of an extraction technique of fish allergens suitable for detection and diagnosis

Ma¹,

Ramesh²,

Li³

et al. 2017

Czech J. Food Sci.

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show abstract

“…Samples (50 lg per well) were mixed 4:1 with loading buffer (2% SDS, 25% glycerol, 14.4 mM b-mercaptoethanol, and 0.1% bromphenol blue in 1 M Tris-HCl, pH 6.8), heated at 100°C for 6 min, electrophoresed in 12% analytical SDS-polyacrylamide gels employing a vertical electrophoresis system (BIO-RAD) according to the manufacturer's recommendations, and either stained with Coomassie Brilliant Blue R-250 (Smith, Cromie, & Stainsby, 1988) or transferred to NC (nitrocellulose membrane, 0.45 nm, Pall-Gelman, USA). For immunoblot, proteins were electrophoretically transferred from the gels to NC by applying a constant current of 8.0 mA/cm 2 for 2 h at room temperature, essentially according to the method of Towbin and Gordon (1984).…”

Section: Protein Electrophoresis and Immunoblottingmentioning

confidence: 99%

The influence of gamma irradiation on the allergenicity of shrimp (Penaeus vannamei)

Hong

Cao

et al. 2007

Journal of Food Engineering

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“…Denaturing protein electrophoresis (SDS-PAGE) was performed according to the method of Laemmli (1970). Samples (50 µg per well) were mixed 4:1 with loading buffer (2% SDS, 25% glycerol, 14.4 mmol/L β-mercaptoethanol, and 0.1% bromphenol blue in 1 mol/L Tris-HCl, pH 6.8), heated at 100 °C for 6 min, electrophoresed in 12% analytical SDS-polyacrylamide gels employing a vertical electrophoresis system (Bio-Rad) according to the manufacturer's recommendations, and either stained with Coomassie Brilliant Blue R-250 (Smith et al, 1988) or transferred to NC (nitrocellulose membrane, 450 nm, Pall-Gelman, USA). For immunoblot, proteins were electrophoretically transferred from the gels to NC by applying a constant current of 8.0 mA/cm 2 for 2 h at room temperature, essentially according to the method of Towbin and Gordon (1984).…”

Section: Electrophoresis and Immunoblottingmentioning

confidence: 99%

Effect of high intensity ultrasound on the allergenicity of shrimp

Lin

Cao

et al. 2006

J. Zhejiang Univ. - Sci. B

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The tropomyosin fraction of shrimp proteins is potentially responsible for allergic reaction in individuals with genetic predisposition to allergy. However, there are no efficient and safe methods to reduce its allergenicity. High intensity ultrasound is known to change the structure of proteins. This study is aimed at assessing high intensity ultrasound's effect on the allergenicity of shrimp allergen. Shrimp and purified shrimp allergen were treated with high intensity ultrasound for 30-180 min. Extracts of treated samples were analyzed by enzyme-linked immunosorbent assay (ELISA) with pool serum of shrimp allergy patients and polyclonal anti-allergen antibodies and by immunoblotting after polyacrylamide gel electrophoresis. Shrimp treated with high intensity ultrasound showed a decrease in allergenicity measured with ELISA. A linear relationship between the immune response induced by treated shrimp allergen and the applied treatment time was observed. The decrease in allergenicity was confirmed by immunoblot assays with shrimp allergic patients serum. Allergenicity of shrimp allergen extracted from treated shrimp was higher than that of purified shrimp allergen with the same treatment time. Gel-filtration HPLC was applied for analysis of shrimp allergen after treatment with high intensity ultrasound. Some fractions were appeared with increasing treatment time. The results suggested that high intensity ultrasound could be used to reduce the allergenicity of shrimp.

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Seeing gel wells well

Cited by 17 publications

References 0 publications

Optimisation of an extraction technique of fish allergens suitable for detection and diagnosis

Optimisation of an extraction technique of fish allergens suitable for detection and diagnosis

The influence of gamma irradiation on the allergenicity of shrimp (Penaeus vannamei)

Effect of high intensity ultrasound on the allergenicity of shrimp

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