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SDS-PAGE Hall of Shame
I found this hilarious website from Rice University and had to share!
The comments are so wonderful. I am currently laughing out loud at my desk xD
https://www.ruf.rice.edu/~bioslabs/studies/sds-page/sdsgoofs.html
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So fucking happy to be done with this
I love staining protein gels or membranes. There’s something so fascinating about seeing proteins with your own eyes. Protein electrophoresis We separate proteins by size by loading protein mixtures onto a vertical gel. When electrical current is applied across the gel, proteins (which are negatively charged) will migrate along with the current, travelling through the gel. The gel contains tiny pores which allow proteins to move through it. Small proteins can move easily through these pores but large proteins will lag and move slower due to their size. Therefore, smaller proteins will travel further and end up towards the bottom of the gel, whereas larger proteins will travel a shorter distance and end up towards the top of the gel. The result is this band pattern, where proteins are arranged in decreasing size from the top of the gel to the bottom.
On the outer edges of the membrane (in blue), we’ve loaded a protein ladder. This is a commercial mixture of proteins which has been stained blue and contains multiple proteins of a range of known sizes. These are run and separated on a protein gel along with our protein mixtures so that we can compare the bands in our protein mixtures (stained in red) with the bands in the protein ladder (blue) and therefore calculate the size of our proteins.
March 2, 2017 >> Mixed, poured, set, loaded, ran, and visualized an SDS-PAGE gel all in one lab today 🔬 we loaded three samples from various previous labs to compare their relative amounts of target protein using SDS-PAGE visualization. I was very proud of our gel and babysat it through the entire 4 hour process.
Glutenin, is consider the major storage proteins in the family of Poaceace which is also consider the main contents in wheat cereal of this family, to commercialize the ancient domesticated cereal like barley in the race of industrial crop and determining the uses of barley flour quality and resemblance of barley elite lines elasticity and dough quality, experimental assay was designed in the Laboratory of Plant Molecular Biology Department of Botany, university of Malakand on 100 elite lines of the barley.to detect High Molecular weight Glutenin protein extracted from the Barley seeds on the basis of Molecular weight, the Glu loci in barley 100 lines which score from the gel of polyacrylamide classify into nine different size (120, 105, 90, 80, 65, 45, 30, 20 and 10 KD) bands which further was classified into five different groups on the basis of variability present in selected barley lines and also in their Molecular weight; absence/presence and intensity of bands were observed. Largest group was observed is group 1 have 41% of elite lines in which 120, 105, 90, 80 and 65 KD molecular weight bands were recorded present. And some of the lines were found unknown which are not related any of these five group respectively, which clearly determine the novel allelic subunit and also show variation from the gene pool in these lines, further evaluation of these lines are needed to track these loci in this cash crop.
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The red palm weevil (Rhynchophorus ferrugineus) is one of the most destructive pests affecting economically important palm species worldwide. The present study investigated the insecticidal potential of the ethanol extract of Cocculus hirsutus and evaluated its effects on antioxidant enzyme activity and protein profiles in R. ferrugineus larvae. Leaves of C. hirsutus were collected and extracted using ethanol, and the chemical constituents of the extract were analysed through liquid chromatography–mass spectrometry (LC-MS). A total of 122 compounds were detected, including biologically active molecules such as hirsutine, gallic acid, resveratrol, ferulic acid, and quercetin derivatives. Larvae of R. ferrugineus were exposed to different concentrations (2–10%) of the plant extract. Higher concentrations (8% and 10%) caused rapid mortality of larvae, while the 2% treatment showed negligible effects and was excluded from subsequent biochemical analyses. Antioxidant enzyme assays revealed a marked reduction in catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and polyphenol oxidase (PPO) activities in treated larvae compared with the control, indicating a disruption of oxidative stress defence mechanisms. SDS-PAGE analysis further demonstrated alterations in protein expression, with control larvae showing multiple protein bands, whereas treated larvae exhibited progressively faint bands at higher extract concentrations. These findings suggest that C. hirsutus ethanol extract induces oxidative stress and protein alterations in R. ferrugineus larvae, contributing to their mortality. The study highlights the potential of C. hirsutus as a promising botanical pesticide for the sustainable management of red palm weevil infestations.
The red palm weevil (Rhynchophorus ferrugineus) is one of the most destructive pests affecting economically important palm species worldwide. The present study investigated the insecticidal potential of the ethanol extract of Cocculus hirsutus and evaluated its effects on antioxidant enzyme activity and protein profiles in R. ferrugineus larvae. Leaves of C. hirsutus were collected and extracted using ethanol, and the chemical constituents of the extract were analysed through liquid chromatography–mass spectrometry (LC-MS). A total of 122 compounds were detected, including biologically active molecules such as hirsutine, gallic acid, resveratrol, ferulic acid, and quercetin derivatives. Larvae of R. ferrugineus were exposed to different concentrations (2–10%) of the plant extract. Higher concentrations (8% and 10%) caused rapid mortality of larvae, while the 2% treatment showed negligible effects and was excluded from subsequent biochemical analyses. Antioxidant enzyme assays revealed a marked reduction in catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and polyphenol oxidase (PPO) activities in treated larvae compared with the control, indicating a disruption of oxidative stress defence mechanisms. SDS-PAGE analysis further demonstrated alterations in protein expression, with control larvae showing multiple protein bands, whereas treated larvae exhibited progressively faint bands at higher extract concentrations. These findings suggest that C. hirsutus ethanol extract induces oxidative stress and protein alterations in R. ferrugineus larvae, contributing to their mortality. The study highlights the potential of C. hirsutus as a promising botanical pesticide for the sustainable management of red palm weevil infestations.
The red palm weevil (Rhynchophorus ferrugineus) is one of the most destructive pests affecting economically important palm species worldwide. The present study investigated the insecticidal potential of the ethanol extract of Cocculus hirsutus and evaluated its effects on antioxidant enzyme activity and protein profiles in R. ferrugineus larvae. Leaves of C. hirsutus were collected and extracted using ethanol, and the chemical constituents of the extract were analysed through liquid chromatography–mass spectrometry (LC-MS). A total of 122 compounds were detected, including biologically active molecules such as hirsutine, gallic acid, resveratrol, ferulic acid, and quercetin derivatives. Larvae of R. ferrugineus were exposed to different concentrations (2–10%) of the plant extract. Higher concentrations (8% and 10%) caused rapid mortality of larvae, while the 2% treatment showed negligible effects and was excluded from subsequent biochemical analyses. Antioxidant enzyme assays revealed a marked reduction in catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and polyphenol oxidase (PPO) activities in treated larvae compared with the control, indicating a disruption of oxidative stress defence mechanisms. SDS-PAGE analysis further demonstrated alterations in protein expression, with control larvae showing multiple protein bands, whereas treated larvae exhibited progressively faint bands at higher extract concentrations. These findings suggest that C. hirsutus ethanol extract induces oxidative stress and protein alterations in R. ferrugineus larvae, contributing to their mortality. The study highlights the potential of C. hirsutus as a promising botanical pesticide for the sustainable management of red palm weevil infestations.
