SDS-PAGE gel electrophoresis v2

2021 ◽  
Author(s):  
Steven J Burgess ◽  
Lynn Doran
Keyword(s):  
Gel Electrophoresis ◽  
Protein Extraction ◽  
Extraction Procedure ◽  
Leaf Tissue ◽  
Leading Edge ◽  
Residual Liquid ◽  
Sample Tube ◽  
Coomassie Blue ◽  
Sds Page ◽  
Pipette Tip

SDS-PAGE gel electrophoresis protocol for analyzing samples from plant leaf tissue via immunofluorescence. In this protocol no Coomassie blue is added to samples, the reason is that this interferes with the fluorescent signal during immunoblot. Instead, samples have already been prepared in Laemmli buffer (minus coomassie, protein extraction procedure), the leading edge of samples can be visualized due to the presence of chlorophyll. Note - When using 15 well, 0.75 mm comb, try to limit the volume loaded to 10 μL to minimize the risk of spillover of protein between wells. - Ensure that accurate volume is pipetted by removing sample stuck to the outside of the pipette tip by wiping the tip on the rim of the sample tube to remove any residual liquid. Literature: http://www.bio-rad.com/webroot/web/pdf/lsr/literature/Bulletin_6040.pdfhttps://www.bio-rad.com/webroot/web/pdf/lsr/literature/10026447.pdf

scholarly journals Evaluation of effective protein extraction procedure to profile petiole of Moringa oleifera

2020 ◽  
Vol 7 (2) ◽  
pp. 214
Author(s):  
Zetty Amirah Zulkifli ◽  
Zaidah Rahmat
Keyword(s):  
Moringa Oleifera ◽  
Protein Extraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Extraction Procedure ◽  
Electrophoretic Pattern ◽  
Antioxidant Property ◽  
Extraction Methods ◽  
Protein Profiling ◽  
Sds Page

Moringa oleifera is widely known as multipurpose tree since all of its parts confer multiple functions. The leaf is highly favourable among consumers while the petiole is mostly wasted. There are numerous studies on the flavonoid and antioxidant property of the stem and twig. However, study on the petiole has never been done. There-upon, this study was conducted to develop protein profiling of the petiole. In this study, 6 different protein extraction methods were tested on the fresh petiole before its protein quantity and quality were checked via Bradford assay and Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) respectively. The in-solution digestion was then done prior to LC-MS/MS analysis. The protein electrophoretic pattern from the SDS-PAGE proves that method 6 using Tris HCl buffer with incorporation of dithiothreitol (DTT) and phenylmethylsulfonyl fluoride (PMSF) confers the best quality of protein. It produced the highest number of visible individual bands compared to other methods. Meanwhile, 93 proteins were successfully identified via LCMS analysis where the protein, signal response and carbohydrate metabolism categories confer the highest percentage. High quality and content of the protein extracted from the petiole including the antioxidant, anticancer and antidiabetic protein identified suggested that consuming this part of the plant could enhance nutrients of human body.

A Study on the Recovery Following Freeze–Thaw Injury in Onion and Chrysanthemum

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 454b-454
Author(s):  
Yiwei Jiang ◽  
Chon C. Lim ◽  
Rajeev Arora
Keyword(s):  
Protein Extraction ◽  
Leaf Tissue ◽  
Fold Increase ◽  
Microsomal Protein ◽  
The Other ◽  
Ion Leakage ◽  
Dendranthema Grandiflora ◽  
Freeze Thaw ◽  
Sds Page ◽  
Leaf Tissues

Onion (Allium cepa L. cv Sweet Sandwich) and Chrysanthemum (Dendranthema grandiflora `Sunny Denise') tissues were used to investigate protein changes associated with recovery from freeze–thaw injury. Medium-sized onions were slowly frozen to either –4 or –9 °C, subsequently thawed, and divided in two halves. One half was used immediately for ion leakage (IL) measurements and total and microsomal protein extraction, whereas the other half was allowed to recover at 6 to 8 °C in the dark for 4 to 5 days. Chrysanthemum leaves were frozen to –3.75 °C, and allowed to recover first at 6 to 8 °C in the dark (1 d) and then under 12-h photoperiod at 18 °C (3–4 d). Results indicate a 1.4- and 2.5-fold higher IL, compared to control, from onion tissues frozen to –4 or –9 °C, respectively. IL in –4 °C-treated tissues was the same as respective control following recovery; however, it was further enhanced to 3.6-fold in –9 °C-treated samples. Chrysanthemum leaf tissue exhibited a 1.6-fold increase in ion leakage following injury, but completely recovered to control levels after 4 to 5 d. SDS-PAGE profiles revealed an absence of a 25-kDa microsomal protein in the injured onion tissues but, its up regulation during recovery only in reversibly injured tissues. Data also indicated an accumulation of 36-kDa soluble protein in chrysanthemum leaf tissues during recovery. Experiments are underway to further characterize these protein changes.

scholarly journals Haemophilus influenzaesubtyping by SDS-PAGE of whole-cell polypeptides

1987 ◽  
Vol 99 (1) ◽  
pp. 179-189 ◽  
Author(s):  
A. J. Paterson ◽  
K. F. Macsween ◽  
T. H. Pennington
Keyword(s):  
Haemophilus Influenzae ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Type B ◽  
Dice Coefficient ◽  
Whole Cell ◽  
Coomassie Blue ◽  
Sds Page

SUMMARYStrains ofHaemophilus influenzaeisolated from patients in N.E. Scotland between 1983 and 1986 have been subtyped by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of whole-cell polypeptides. Gels were stained with Coomassie blue and polypeptide profiles were analysed using the Dice coefficient of similarity.Type b strains were all closely related, the 19 strains analysed being grouped at a 90% similarity level into one large (13 strains) and one small (3 strains) cluster with 3 strains being ungrouped. Thirty-six non-typable, epidemiologically unrelated strains were subtyped; one pair of strains had indistinguishable polypeptide profiles. The polypeptide profiles of the remaining strains showed much heterogeneity, although groups of strains isolated from the same patient over short periods showed indistinguishable profiles.

Detección inmunoquímica de la adulteración de chorizo de cerdo con proteínas de soja

1998 ◽  
Vol 4 (4) ◽  
pp. 257-262 ◽  
Author(s):  
A.F. González-Córdova ◽  
A.M. Calderón de la Barca ◽  
M. Cota ◽  
B. Vallejo-Córdoba
Keyword(s):  
Gel Electrophoresis ◽  
Protein Extraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Electrophoretic Pattern ◽  
Soy Flour ◽  
Nacl Solution ◽  
Standard Curve ◽  
Sds Page ◽  
Polyclonal Antisera

Rabbit polyclonal antisera were produced against soy flour proteins extracted with 0.5 M NaCl solution and purified by affinity chromatography to detect adulteration in pork chorizo (sausage) with an enzyme-linked immunoabsorbent assay (ELISA). To detect adulteration the following saline extracts were prepared: 100% pork and 100% soy chorizo and mixtures of these two pro ducts (90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, 10:90), and extracts of two commercial chorizos labeled as pork. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that the salt solution was the most suitable for protein extraction compared to water and a 1% SDS solution. The commercial chorizos did not have the characteristic electrophoretic pattern for pork chorizos. The antibodies were specific for soy and its products, as there was no response to other vegetable and animal proteins by gel immunodiffusion assay or ELISA. The standard curve obtained with the ELISA results of the chorizo mixtures gave a correlation coefficient of r2 = 0.988. The two commercial chorizo values resulted in a 32 and 40% soy substitution. Total time for ELISA was optimized to 4 h, and 2 h if the plates were previously activated with the antigen. Variation coefficients of ELISA readings for replicates of the same extract in one plate and variation of different assays on different days were 2.3 and 8% respectively.

Heterozygous Abnormal Fibrinogen Osaka III with the Replacement of γ Arginine-275 by Histidine Has an Apparently Higher Molecular Weight γ-Chain Variant

1992 ◽  
Vol 68 (05) ◽  
pp. 534-538 ◽  
Author(s):  
Nobuhiko Yoshida ◽  
Shingi Imaoka ◽  
Hajime Hirata ◽  
Michio Matsuda ◽  
Shinji Asakura
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Tetraacetic Acid ◽  
Fibrin Monomer ◽  
Sds Page ◽  
Thrombotic Tendency ◽  
Chain Variant

SummaryCongenitally abnormal fibrinogen Osaka III with the replacement of γ Arg-275 by His was found in a 38-year-old female with no bleeding or thrombotic tendency. Release of fibrinopeptide(s) by thrombin or reptilase was normal, but her thrombin or reptilase time in the absence of calcium was markedly prolonged and the polymerization of preformed fibrin monomer which was prepared by the treatment of fibrinogen with thrombin or reptilase was also markedly defective. Propositus' fibrinogen had normal crosslinking abilities of α- and γ-chains. Analysis of fibrinogen chains on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the system of Laemmli only revealed the presence of abnormal γ-chain with an apparently higher molecular weight, the presence of which was more clearly detected with SDS-PAGE of fibrin monomer obtained by thrombin treatment. Purified fragment D1 of fibrinogen Osaka III also seemed to contain an apparently higher molecular weight fragment D1 γ remnant on Laemmli gels, which was digested faster than the normal control by plasmin in the presence of [ethy-lenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA).

Identifikasi Daging Tikus Pada Produk Baso Dengan Metode Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE)

2018 ◽  
Vol 26 (2) ◽  
pp. 058
Author(s):  
Anna P. Roswiem ◽  
Triayu Septiani
Keyword(s):  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Sds Page

<em>Bahan<strong> </strong>baku untuk membuat baso adalah daging hewan, pada umumnya dari daging sapi, ayam, ikan dan babi. Di beberapa daerah di Indonesia terjadi kasus baso tikus. Tujuan penelitian ini adalah menguji ada tidaknya kandungan daging tikus pada produk baso yang dijual di pasar Cempaka Putih-Kecamatan Kramat Jakarta Pusat dan di pedagang baso atau mie baso di sekitar kampus Universitas YARSI Jakarta. Daging adalah protein salah satu metode untuk mengidentifikasi protein adalah metode Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE).<strong> </strong>Hasil penelitian menunjukkan bahwa dari 6 sampel baso terindikasi ada 2 sampel baso dengan nomor 1 dan 5 yang dibuat dari campuran daging sapi dan tikus; ada 1 sampel baso dengan nomor 6 yang terbuat dari daging tikus; dan 2 sampel baso dengan nomor 2 dan 3 yang terbuat dari campuran sapi  dan babi, dan hanya 1 sampel baso dengan nomor sampel 4 yang benar-benar terbuat dari daging sapi.</em>

scholarly journals Protocol Optimization of Proteomic Analysis of Korean Ginseng (Panax ginseng Meyer)

Separations ◽  
2021 ◽  
Vol 8 (4) ◽  
pp. 53
Author(s):  
Clarissa Braccia ◽  
Bhakti Prinsi ◽  
Mara Colzani ◽  
Alessandra A. Altomare ◽  
Luca Espen ◽  
...  
Keyword(s):  
Panax Ginseng ◽  
Proteomic Analysis ◽  
Metabolic Pathways ◽  
Extraction Procedure ◽  
Biotic Stresses ◽  
Phenol Extraction ◽  
Ms Analysis ◽  
Sds Page ◽  
Protein Properties ◽  
Korean Ginseng

The benefits of ginseng have been mainly attributed to its triterpenoids, called ginsenosides. Recent genome sequencing of the Panax ginseng has paved the way for in-depth proteomic studies of this medicinal plant. The current study was conducted to deepen the proteomic information on the root proteome of Korean ginseng. Proteomic workflow was optimized by testing two different strategies, characterized by the phenol extraction procedure, the presence or the absence of SDS-PAGE fractionation step, and nano-scale liquid chromatographic tandem mass spectrometry (nLC-MS/MS) analysis. The results highlighted an evident improvement of proteome extraction by the combination of phenol extraction with SDS-PAGE before the nLC-MS/MS analysis. In addition, a dramatic impact of the steaming process (the treatment to produce red ginseng from ginseng) on protein properties was observed. Overall, the analyses of Korean ginseng permitted the characterization of a total of 2412 proteins. A large number of identified proteins belonged to the functional categories of protein and carbon/energy metabolism (22.4% and 14.6%, respectively). The primary and secondary metabolisms are major metabolic pathways, which emerged from the proteomic analysis. In addition, a large number of proteins known to play an important role in response to (a)biotic stresses were also identified. The current proteomic study not only confirmed the previous transcriptomic and proteomic reports but also extended proteomic information, including the main metabolic pathways involved in Korean ginseng.

scholarly journals Characterizing Wool Keratin

2009 ◽  
Vol 2009 ◽  
pp. 1-5 ◽  
Author(s):  
Jeanette M. Cardamone ◽  
Alberto Nuñez ◽  
Rafael A. Garcia ◽  
Mila Aldema-Ramos
Keyword(s):  
Hydrogen Peroxide ◽  
Gel Electrophoresis ◽  
Water Soluble ◽  
Alkaline Solutions ◽  
Biodegradable Material ◽  
Alkaline Hydrogen Peroxide ◽  
Cortical Cells ◽  
Sds Page ◽  
Fibrous Matrices ◽  
Wool Keratin

Keratin from wool is a reactive, biocompatible, and biodegradable material. As the biological structural component of skin (soft keratins) and of nails, claws, hair, horn, feathers, and scales (hard keratins) pure keratin comprises up to 90% by weight of wool. Wool was treated in alkaline solutions to extract from 68% to 82% keratin within 2 to 5 hours of exposure at . The keratin products were water-soluble and were confirmed to contain intermediate filament and microfibrillar component-proteins of fractured, residual cuticle, and cortical cells. Oxidation of wool by peroxycarboximidic acid in alkaline hydrogen peroxide produced keratin products with distinct microcrystalline structures: descaled fibers, fibrous matrices, and lyophilized powders. Morphology and confirmation of peptide functionality were documented by SEM, Amino Acid Analysis, SDS-PAGE gel electrophoresis, MALDI-TOF/TOF, and FTIR analyses. The reactivity of keratin from wool models the reactivity of keratin from low-value sources such as cattle hair.

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