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1. A systematic approach to quantitative Western blot analysis

Author:
Pillai-Kastoori, Lakshmi; Schutz-Geschwender, Amy R.; Harford, Jeff A.
Source:
Analytical biochemistry 2020 v.593 pp. 113608
ISSN:
0003-2697
Subject:
Western blotting,   etc   ; antibodies; electrophoresis; image analysis; proteins;     Show all 5 Subjects
Abstract:
... Attaining true quantitative data from WB requires that all the players involved in the procedure are quality controlled including the user. Appropriate protein extraction method, electrophoresis, and transfer of proteins, immunodetection of blotted protein by antibodies, and the ultimate step of imaging and analyzing the data is nothing short of a symphony. Like with any other technology in life-s ...
DOI:
10.1016/j.ab.2020.113608

2. Application of Toxoplasma gondii GRA15 peptides in diagnosis and serotyping

Author:
Li, Runli; Ma, Yeting; Li, Jin; Zhou, Penglai; Zheng, Fuguo; Liu, Qing; Gao, Wenwei
Source:
Microbial pathogenesis 2020 v.143 pp. 104168
ISSN:
0882-4010
Subject:
Western blotting,   etc   ; Toxoplasma gondii; antibodies; genes; genotyping; peptides; proteins; serodiagnosis;     Show all 8 Subjects
Abstract:
... Previous studies showed that three clonal strain types (I, II, and III) of Toxoplasma gondii can be distinguished using serotyping based on a series of polymorphic proteins. However, to establish a systematic serotyping method with higher resolution even being equal to that of genotyping, more specific peptide markers are needed. The objective of the present study was to determine the possibility ...
DOI:
10.1016/j.micpath.2020.104168

3. PHF23 negatively regulates the autophagy of chondrocytes in osteoarthritis

Author:
Maimaitijuma, Talatibaike; Yu, Jia-Hong; Ren, Ya-Li; Yang, Xin; Liu, Heng; Meng, Zhi-Chao; Wang, Rui; Cui, Yun-Peng; Wu, Hao; Pan, Li-Ping; Jiao, Yang; Chen, Ying-Yu; Cao, Yong-Ping
Source:
Life sciences 2020 v.253 pp. 117750
ISSN:
0024-3205
Subject:
Western blotting,   etc   ; autophagosomes; cell viability; chloroquine; chondrocytes; collagen; elderly; interleukins; lysosomes; mitophagy; osteoarthritis; phosphorylation; staining; surgery;     Show all 14 Subjects
Abstract:
... Osteoarthritis (OA) is the main cause of disability and joint replacement surgery in the elderly. As a crucial cell survival mechanism, autophagy has been reported to decrease in OA. PHF23 is a new autophagy inhibitor which was first reported by us previously. This study aimed to explore the anti-autophagic mechanism of PHF23 to make it a possible therapeutic target of OA.Lentiviral vectors specif ...
DOI:
10.1016/j.lfs.2020.117750

4. Screening anti-TMV agents targeting tobacco mosaic virus helicase protein

Author:
Wang, Dongmei; Huang, Maoxi; Gao, Di; Chen, Kai; Xinxie,; Xu, Weiming; Li, Xiangyang
Source:
Pesticide biochemistry and physiology 2020 v.166 pp. 104449
ISSN:
0048-3575
Subject:
Western blotting,   etc   ; Nicotiana benthamiana; Tobacco mosaic virus; antiviral agents; binding sites; computer simulation; dissociation; enzymes; screening;     Show all 9 Subjects
Abstract:
... Tobacco mosaic virus helicase (TMV-Hel) plays important roles in viral multiplication. TMV-Hel is a potential target of anti-TMV agents. Our previous studies expressed and purified TMV-Hel as target protein for cytosinpeptidemycin. In this study, we preform molecular docking to study the binding sites of commercial antiviral agents with TMV-Hel. Then we verify the interactions between the potentia ...
DOI:
10.1016/j.pestbp.2019.07.017

5. A method to study protein biomarkers in saliva using an automated capillary nano-immunoassay platform (Wes™)

Author:
Reyes-Serratos, Eduardo; Marcet-Palacios, Marcelo; Rosenfield, David; Ritz, Thomas; Befus, A. Dean
Source:
Journal of immunological methods 2020 v.479 pp. 112749
ISSN:
0022-1759
Subject:
Western blotting,   etc   ; algorithms; antibodies; automation; biomarkers; calcium-binding proteins; molecular weight; saliva;     Show all 8 Subjects
Abstract:
... Traditional immunoprobing techniques like Western blot continue to play a crucial role in the discovery and validation of biomarkers. This technique suffers from several limitations that affect reproducibility and feasibility for large-scale studies. Modern immunoprobing techniques have addressed several of these limitations. Here we contrast the use of Western blot and an automated capillary nano ...
DOI:
10.1016/j.jim.2020.112749

6. Fast immunoassay for microfluidic western blotting by direct deposition of reagents onto capture membrane

Author:
Arvin, Natalie E.; Dawod, Mohamed; Lamb, Don T.; Anderson, Jon P.; Furtaw, Michael D.; Kennedy, Robert T.
Source:
Analytical methods 2020 v.12 no.12 pp. 1606-1616
ISSN:
1759-9679
Subject:
Western blotting,   etc   ; antibodies; automation; detection limit; electrophoresis; immunoassays; microchip technology; tubulin;     Show all 8 Subjects
Abstract:
... Western blotting is a widely used protein assay platform, but the technique requires long analysis times and multiple manual steps. Microfluidic systems are currently being explored for increased automation and reduction of analysis times, sample volumes, and reagent consumption for western blots. Previous work has demonstrated that proteins separated by microchip electrophoresis can be captured o ...
DOI:
10.1039/d0ay00207k

7. HBXIP: a potential prognosis biomarker of colorectal cancer which promotes invasion and migration via epithelial-mesenchymal transition

Author:
Wang, Xuanyu; Feng, Qi; Yu, Hongnv; Zhou, Xingzhi; Shan, Changliang; Zhang, Qinggao; Liu, Shuangping
Source:
Life sciences 2020 v.245 pp. 117354
ISSN:
0024-3205
Subject:
Western blotting,   etc   ; biomarkers; colorectal neoplasms; hepatitis B; metastasis; patients; prognosis; tissues;     Show all 8 Subjects
Abstract:
... Hepatitis B X-interacting protein (HBXIP) is highly expressed in many cancers, but the correlation between the expression of HBXIP and the clinical significance and underlying molecular mechanisms in colorectal cancer (CRC) is still unclear. We selected 186 specimens from CRC patients for analyzing the relationship between the expression of HBXIP and the clinical-pathological features by immunohis ...
DOI:
10.1016/j.lfs.2020.117354

8. Progesterone induces sperm release from oviductal epithelial cells by modifying sperm proteomics, lipidomics and membrane fluidity

Author:
Ramal-Sanchez, Marina; Bernabo, Nicola; Tsikis, Guillaume; Blache, Marie-Claire; Labas, Valerie; Druart, Xavier; Mermillod, Pascal; Saint-Dizier, Marie
Source:
Molecular and cellular endocrinology 2020 v.504 pp. 110723
ISSN:
0303-7207
Subject:
Western blotting,   etc   ; cattle; confocal microscopy; epithelial cells; insemination; lipids; membrane fluidity; oviducts; progesterone; proteins; proteomics; spermatozoa;     Show all 12 Subjects
Abstract:
... The sperm reservoir is formed after insemination in mammals, allowing sperm storage in the oviduct until their release. We previously showed that physiological concentrations of progesterone (P4) trigger in vitro the sperm release from bovine oviductal epithelial cells (BOECs), selecting a subpopulation of spermatozoa with a higher fertilizing competence. Here, by using Western-Blot, confocal micr ...
DOI:
10.1016/j.mce.2020.110723

9. A sensitive and reversible staining of proteins on blot membranes

Author:
Wang, Jun-Ling; Zhao, Li; Li, Mei-Qi; Chen, Wei-Guang; Xu, Chao-Jin
Source:
Analytical biochemistry 2020 v.592 pp. 113579
ISSN:
0003-2697
Subject:
Western blotting,   etc   ; Congo red; ambient temperature; biotechnology; chemiluminescence; computer simulation; enzyme substrates; fluorides; peroxidase; proteins; staining;     Show all 11 Subjects
Abstract:
... A modified, sensitive and reversible method for protein staining on nitrocellulose (NC) and polyvinylidine fluoride (PVDF) membranes was developed in Western blotting. The method employed Congo red staining to visualize proteins on different blot membranes. Staining of proteins with Congo red dye is more faster procedures. According to the experimental results, approximate 20 ng proteins could be ...
DOI:
10.1016/j.ab.2020.113579

10. Parkin overexpression attenuates Aβ-induced mitochondrial dysfunction in HEK293 cells by restoring impaired mitophagy

Author:
Wang, Hongmei; Zhang, Ting; Ge, Xuhua; Chen, Jingjiong; Zhao, Yuwu; Fu, Jianliang
Source:
Life sciences 2020 v.244 pp. 117322
ISSN:
0024-3205
Subject:
Western blotting,   etc   ; Alzheimer disease; dyes; membrane potential; mitochondria; mitochondrial membrane; mitophagy; therapeutics;     Show all 8 Subjects
Abstract:
... Mitochondrial dysfunction is an early prominent feature of Alzheimer's disease (AD). In the present study, we sought to investigate whether defective mitophagy is tightly related to amyloid-β (Aβ)-induced mitochondrial dysfunction.Immunofluorescence, western blot and transmission electron microscopy were used to examine mitophagy. Mitochondrial membrane potential was assessed using the JC-1 dye. M ...
DOI:
10.1016/j.lfs.2020.117322

11. Uterine and placental specific localization of AQP2 and AQP8 is related with changes of serum progesterone levels in pregnant queens

Author:
Ferré-Dolcet, Lluís; Yeste, Marc; Vendrell, Meritxell; Rigau, Teresa; Rodríguez-Gil, Joan Enric; del Alamo, Maria Montserrat Rivera
Source:
Theriogenology 2020 v.142 pp. 149-157
ISSN:
0093-691X
Subject:
Western blotting,   etc   ; aquaporins; blood serum; cytoplasm; endometrium; epithelium; immunohistochemistry; myometrium; plasma membrane; pregnancy; progesterone; trophoblast;     Show all 12 Subjects
Abstract:
... Aquaporins play vital roles in reproductive physiology. This study evaluates the expression and localization dynamics of AQP1, AQP2, AQP3 and AQP8 in the endometrium and placental transference zone during pregnancy in queens by means of immunohistochemistry and Western blot. Animals were distributed into six groups: non-pregnant queens with low levels of serum progesterone (P4), non-pregnant anima ...
DOI:
10.1016/j.theriogenology.2019.09.049

12. Ezrin T567 phosphorylation regulates migration and invasion of ectopic endometrial stromal cells by changing actin cytoskeleton

Author:
Chen, Chaolu; Ye, Chaoshuang; Xia, Jianmei; Zhou, Yong; Wu, Ruijin
Source:
Life sciences 2020 v.254 pp. 117681
ISSN:
0024-3205
Subject:
Western blotting,   etc   ; actin; cell movement; endometrium; fluorescent antibody technique; immunohistochemistry; microfilaments; phosphorylation; pseudopodia; signal transduction; stromal cells; therapeutics;     Show all 12 Subjects
Abstract:
... The enhanced ability of endometrial cell migration and invasion is the foundation for formation of ectopic lesions in endometriosis. Ezrin has been reported to regulate cell motility by remodeling the cytoskeleton. However, little is known about the mechanisms through which ezrin remodels the cytoskeleton and cell structure to promote cell motility in endometriosis.In our study, expression and dis ...
DOI:
10.1016/j.lfs.2020.117681

13. Effects of inhalation of sevoflurane at different concentrations on TRPV1 in airways of rats at different developmental stages

Author:
Liu, Dexing; Yuan, Jie; Fei, Xia; Zhu, Yuhang; Zhou, Yannan; Zhang, Chao; Dong, Liang; Zhu, Zhaoqiong
Source:
Life sciences 2020 v.249 pp. 117472
ISSN:
0024-3205
Subject:
Western blotting,   etc   ; anesthesia; breathing; calcitonin gene-related peptide; developmental stages; lungs; oxygen; sevoflurane; transient receptor potential vanilloid channels;     Show all 9 Subjects
Abstract:
... AimDetermine changes in the expressions of the ion channel—TRPV1—and neuropeptides—NKA, NKB, calcitonin gene-related peptide (CGRP), and SP—in 14-, 21-, and 42-day-old rats after inhaling 1.5% and 2.6% sevoflurane.A small in-house inhalation anesthesia chamber was designed to allow 14-, 21-, and 42-day-old rats inhale 1.5% and 2.6% sevoflurane, and rats in the control group inhaled carrier gas(1 L ...
DOI:
10.1016/j.lfs.2020.117472

14. Protein extraction from Saccharomyces cerevisiae at different growth phases

Author:
Mukherjee, Madhumathan; Nandi, Ankita; Chandra, Krishna; Saikia, Surjya Kumar; Jana, Chandan Kumar; Das, Nilanjana
Source:
Journal of microbiological methods 2020 v.172 pp. 105906
ISSN:
0167-7012
Subject:
Western blotting,   etc   ; Saccharomyces cerevisiae; alkali treatment; bioactive properties; cell walls; developmental stages; genome; glass; hydrophilicity; models; proteins; proteomics; sonication;     Show all 13 Subjects
Abstract:
... Saccharomyces cerevisiae is an established model organism with a well characterized genome. However, this model presents a unique problem due to a very resistant cell wall which develops in the late stationary phase resulting in sub-optimal extraction of proteins from such cells using majority of the cell lysis protocols. In this study, several methods from the literature with modifications thereo ...
DOI:
10.1016/j.mimet.2020.105906

15. Notch signal deficiency alleviates hypertrophic scar formation after wound healing through the inhibition of inflammation

Author:
He, Ting; Bai, Xiaozhi; Jing, Jing; Liu, Yang; Wang, Hongtao; Zhang, Wanfu; Li, Xiaoqiang; Li, Yan; Wang, Luxu; Xie, Songtao; Hu, Dahai
Source:
Archives of biochemistry and biophysics 2020 v.682 pp. 108286
ISSN:
0003-9861
Subject:
Western blotting,   etc   ; chemotaxis; collagen; cytokines; fibrosis; inflammation; knockout mutants; macrophages; mice; models; reverse transcriptase polymerase chain reaction; staining; tissue repair;     Show all 13 Subjects
Abstract:
... Pathological scar is a common complication after wound healing. One of the most important factors that affects scar formation is inflammation. During this process, macrophages play a critical role in the wound healing process, as well as in scar formation. Notch signaling is reported to participate in inflammation and fibrosis; however, whether it affects scar formation is still unclear. In this s ...
DOI:
10.1016/j.abb.2020.108286

16. An easy and simple separation method for Fc and Fab fragments from chicken immunoglobulin Y (IgY)

Author:
Zhou, Xin; Wang, Yanru; Ahn, Dong Uk; Cai, Zhaoxia
Source:
Journal of chromatography 2020 v.1141 pp. 122011
ISSN:
1570-0232
Subject:
Western blotting,   etc   ; agarose; ammonium sulfate; buffers; chickens; chromatography; hydrolysis; immunoglobulin Y; ion exchange; pH; papain; purification methods; sodium chloride;     Show all 13 Subjects
Abstract:
... Antigen-binding (Fab) and crystallizable (Fc) fragments are the active components of yolk immunoglobulin (IgY), which have been widely used in the pharmaceutical field. However, the common purification methods for the Fab and Fc fragments use combinations of multi-columns are complex and time-consuming. The objective of this study was to improve the separation efficiency of the Fab and Fc fragment ...
DOI:
10.1016/j.jchromb.2020.122011

17. Optimization of blocking conditions for fluorescent Western blot

Author:
Cui, Yan
Source:
Analytical biochemistry 2020 v.593 pp. 113598
ISSN:
0003-2697
Subject:
Western blotting,   etc   ; antibodies; bovine serum albumin; cost effectiveness; fish; fluorescence; gelatin; milk; polysorbates; polyvinylpyrrolidone; sodium dodecyl sulfate;     Show all 11 Subjects
Abstract:
... Blocking conditions are critical for reducing background and non-specific signals in Western blot (WB). For fluorescent WB, however, blockers may bring in additional autofluorescence, and optimal blocking conditions have been less well established. Here, the autofluorescence of Tween 20 is clarified, which is negligible when imaged wet, but could be significant when Tween 20 is dried on blots. In ...
DOI:
10.1016/j.ab.2020.113598

18. Development of Recombinant Immunoglobulin G-Binding Luciferase-Based Signal Amplifiers in Immunoassays

Author:
Eom, Soomin; Bae, Yoonji; Kim, Sunghwan; Choi, Hyukjun; Park, Jongnam; Kang, Sebyung
Source:
Analytical chemistry 2020 v.92 no.7 pp. 5473-5481
ISSN:
1520-6882
Subject:
Western blotting,   etc   ; animal use replacement; antibodies; chemical bonding; enzyme-linked immunosorbent assay; immunoglobulin G; luciferase; mice; rabbits;     Show all 9 Subjects
Abstract:
... In general immunoassays, secondary antibodies are covalently linked with enzymes and bind to the Fc region of target-bound primary antibodies to amplify signals of low-abundant target molecules. The antibodies themselves are obtained from large mammals and are further modified with enzymes. In this study, we developed novel recombinant immunoglobulin G (IgG)-binding luciferase-based signal amplifi ...
DOI:
10.1021/acs.analchem.0c00222

19. JAK2-STAT5 signaling is insensitive to porcine growth hormone (pGH) in hepatocytes of neonatal pig

Author:
Yu-Jiang, Yang; Xin, Zheng; Hai-Nan, Lan
Source:
Animal cells and systems 2020 v.24 no.2 pp. 69-78
ISSN:
2151-2485
Subject:
Western blotting,   etc   ; cell nucleus; hepatocytes; models; non-specific protein-tyrosine kinase; precipitin tests; signal transduction; somatotropin; somatotropin receptors; swine; transactivators;     Show all 11 Subjects
Abstract:
... Porcine growth hormone (pGH) is most important hormone which is involved in the growth and development of pig. However, a series of studies have indicated that neonatal pig is insensitive to pGH; the reason for this phenomenon is still not fully understood. In this work, we try to investigate this issue from the angle of intracellular signaling induced by pGH. In the present study, porcine hepatoc ...
DOI:
10.1080/19768354.2020.1735518

20. A natural anthraquinone derivative shikonin synergizes with AZD9291 against wtEGFR NSCLC cells through reactive oxygen species-mediated endoplasmic reticulum stress

Author:
Hu, Xiu; Zhang, Zuo-yan; Wu, Lin-wen; Zeng, Ling-hui; Chen, Hui; Zhu, Hua-jian; Zhang, Jian-kang; Shao, Jiaan; Zhang, Chong; Li, Yang-ling; Lin, Neng-ming
Source:
Phytomedicine 2020 v.68 pp. 153189
ISSN:
0944-7113
Subject:
Western blotting,   etc   ; anthraquinones; antineoplastic activity; apoptosis; endoplasmic reticulum stress; lung neoplasms; mutation; oxygen; patients; propidium; shikonin; staining; survival rate; therapeutics;     Show all 14 Subjects
Abstract:
... NSCLC is the major type of lung cancer and the survival rates of NSCLC patients remain low. AZD9291 is a third-generation EGFR-TKI and approved to treat NSCLC patients harboring EGFR T790M mutation and common targetable activating EGFR mutations, but it has a limited effect for wtEGFR NSCLC.The current study investigated whether shikonin could enhance the antitumor effect of AZD9291 in wtEGFR NSCL ...
DOI:
10.1016/j.phymed.2020.153189