Science method
Western Blot - Science method
Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes.
Publications related to Western Blot (5,665)
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La melatonina regula la estacionalidad reproductiva en la
especie ovina mediante su unión a receptores de membrana,
como MT1. El gen que codifica para este receptor
(MTNR1A) presenta varios polimorfismos, entre ellos el
RsaI (g.17355458C>T), relacionado con una menor estacionalidad.
A nivel nuclear, el RORα (retinoic acid-related
orphan receptor al...
The human interferon-induced transmembrane (IFITM) proteins inhibit the fusion of a broad spectrum of enveloped viruses, both when expressed in target cells and when present in infected cells. Upon expression in infected cells, IFITMs incorporate into progeny virions and reduce their infectivity by a poorly understood mechanism. Since only a few En...
Objectives
To explore the role of dual-specific phosphatase 1 (DUSP1) in the proliferation, migration and invasion of pancreatic cancer (PC).
Methods
TCGA and GTEx databases were used to investigate the relationship between DUSP1 expression and prognosis of PC patients. Expression efficiency of DUSP1 was validated by qPCR and western blotting. The...
Protocol for performing an SDS-PAGE Western blot analysis using the BioRad Criterion Blotter system.
Preparation of Nuclear Extracts using high salt and sonication for western blot applications
We got a great result to detect alpha-synuclein and GCase protein using this WB protocol
The recombinant antibodies AF641, AI954 and AW954 detect different markers of the endoplasmic reticulum by western blot.
Varón de 42 años, ingresa por el servicio de emergencias presentando dificultad respiratoria progresiva, malestar general y fiebre, se decide hospitalizar en esta institución con el diagnóstico de Insuficiencia Respiratoria Aguda por neumonía viral, se requirió descartar COVID-19 por hallazgos sugestivos y se obtuvo prueba molecular negativa. Media...
Relative protein quantification is a well-established technique in the vast majority of molecular biology laboratories. However, western blot standard protocols may not detect proteins of certain sizes. When the molecular weight of protein of interest is out of the 10–250 kDa range, its migration through the gel or transfer to the membrane is compr...
Background. Several studies suggest that the pleiotropic properties of betanin may interfere with different signaling pathways. Our previous studies on human hepatocytes showed that betanin activated the nuclear factor erythroid-2-related factor 2 (Nrf2) signaling pathway. To further understand the exact mechanism of action of betanin, we evaluated...
In our previous study of Hu sheep hair follicles, we found that CRABP2 was highly expressed in DPCs, which suggested that CRABP2 may influence the number of DPCs. In the present study, we aimed to understand the effect of CRABP2 in Hu sheep dermal papilla cells (DPCs). First, we explored the influence of CRABP2 on the ability of Hu sheep DPCs’ prol...
REST (RE1-Silencing Transcription factor) es un factor de transcripción con dedos de zinc que reprime a sus blancos transcripcionales a través de su interacción con la secuencia RE1 (Restrictive Element 1). Aunque algunos metales como el cadmio pueden alterar la función de proteínas al competir con el zinc, no se han realizado estudios a nivel mole...
The protocol describes cell surface biotinylation to identify plasma membrane localized proteins in cell culture via western blotting.
ELISA or Western blot is known as a basic technique to be used for measurement of intracellular proteins, but in some cases, they cannot overcome problems such as normalization between samples or extraneous costs for required commercial kits. In order to address this problem, we developed a rapid and effective method (a hybrid of Western blot and E...
Comparison and assessment between Western blotting and Enzyme-linked Immunosorbent Assay (ELISA) regarding protein analysis.
Subsequently to the publication of this paper, the authors' noticed that the same β‑actin control bands were inadvertently used in the western blots shown in Figs. 1A and 2A. After having examined their original data, the authors realized the control bands were chosen incorrectly for Fig. 1, but were able to identify the data that should have been...
Es un inmunoensayo simple y rápido. Requiere la aplicación de una pequeña cantidad de muestra directamente sobre una membrana polivinilo PVDF, en la que se deposita antigeno total de DENV y posteriormente se expone a suero humano diluido en TBS-T 1:200, para posteriormente, ser detectado con anticuerpo IgG anti human marcado con fosfatasa alcalina...
Protocol for detecting Rab7a phosphorylation at S72 by LRRK1 and LRRK1 mutants in HEK 293T cells.
This protocol details list of antibodies used for immunohistochemistry and western blot.
Western Blotting is a technique for the immunodetection of proteins using antibodies with fluorescent or chemiluminescent detection.
Western blotting (immunoblotting) is a powerful and commonly used technique that is capable of detecting or semiquantifying an individual protein from complex mixtures of proteins extracted from cells or tissues. The history surrounding the origin of western blotting, the theory behind the western blotting technique, a comprehensive protocol and th...
Darbeli Elektromanyetik Alan (PEMF) düşük frekanslı elektromanyetik alan olup son yıllarda klinik araştırmalarda tedavi amaçlı uygulanmasına yönelik çalışmalar hız kazanmıştır. Farklı frekans, yoğunluk, dalga boyu ve sürelerde kematerapötik ilaçlarla birlikte uygulanan PEMF maruziyetinin beyin kanseri hücreleri dahil çeşitli kanser hücrelerinde apo...
Background: To explore the biological effects of CASC11 on aggravating diabetic nephropathy (DN) by regulating FoxO1 (forkhead transcription factor O1). Methods: Serum levels of CASC11 and FoxO1 in DN patients were detected. The possibility of CASC11 in predicting the onset of DN was analyzed by depicting ROC curves. Correlation between CASC11 and...
This protocol describes in vitro kinase assay, in which PI3Kc is tested for phosphorylation by TBK1 and ULK1 complex either by western blotting (S29 on ATG14L subunit) or mass spectrometry analysis.
This protocol describes how to detect alpha-synuclein in protein derived from STC-1 cells by western blot using DAB/peroxidase or ECL to visualise the bands.
Introduction
The aim of this study was to investigate the relationship between ferroptosis and myocardial fibrosis in atrial fibrillation (AF).
Material and methods
The patient’s left atrial appendage tissue was obtained in cardiac surgery. Sirius red staining, Prussian blue and Western blot were used to detect staining to assess histological fibr...
circRNAs play an important role in the progression of osteoarthritis (OA). Therefore, we aimed to reveal the mechanism of action of circRNA-ZCCHC14 in OA. OA animal and cell models were constructed, and clinical samples were collected. The expression of circRNA-ZCCHC14 and miR-181a was detected by RT‒qPCR. The chondrogenic differentiation ability o...
Background:
Tumor-produced high molecular weight insulin-like growth factor-II (big insulin-like growth factor-II) is considered to cause non-islet cell tumor hypoglycemia. This paper presents a case of surgically resected retroperitoneal liposarcoma that produced big insulin-like growth factor-II.
Case presentation:
Here, we report the case of...
Se presenta el caso de un paciente varón de 28 años, heterosexual, con VIH negativo que desarrolló erupción cutánea intensa caracterizada por la presencia de vesículas, pústulas y pápulas umbilicadas confluentes en el pene y región púbica con escasa diseminación en el resto del cuerpo, sin adenopatía ni manifestaciones sistémicas, cuatro días despu...
The recombinant antibodies RB393, RB394 and RB395 do not detect by western blot the AplA protein.
Characterizing macromolecular interactions is essential for understanding cellular processes, yet most methods currently used to detect protein interactions from cells are qualitative. Here, we introduce the native holdup (nHU) approach to estimate equilibrium binding constants of protein interactions directly from cell extracts. Compared to other...
Background
At present, the function of numerous individual regulatory elements has been characterized extensively, but the combined effect of various cis-regulatory elements on gene expression in mammalian cells has been less discussed.
Results
In the present study, we examined the function of the combined effect of various cis-regulatory elements...
Objective
To detect the changes in Nel-like 2 (NELL2) in cerebrospinal fluid (CSF) in the outcome of tuberculous meningitis (TBM) patients and to initially evaluate its potential as a marker.
Methods
We collected the clinical data of patients with suspected TBM in the First People’s Hospital of Zunyi from November 2017 to January 2021 and retained...
Uvod: Lajm neuroborelioza je infektivni poremećaj centralnog i/ili
perifernog nervnog sistema uzrokovan ubodom krpelja roda Ixodes
rici, zaraženog spirohetom Borrelia burgdorferi sensu lato (u
Evropi). Bolest se manifestuje kao meningitis, encefalitis,
meningoradikulitis, vaskulitis, pareza facijalnog nerva i bolne
radikulopatije.
Prikaz slučaja: P...
Protocol to detect ATP13A2 and ATP13A3 via Western Blotting.
Li Y, Wang Q, Wang D, Fu W. Cancer Manag Res. 2021;13:5547–5557.
At the authors request, the Editor and Publisher of Cancer Management and Research wish to retract the published article. Concerns were raised regarding the alleged duplication of western blot and flow cytometry images in Figures 2 and 3 with comparable images from Liu et al, 2021 (ht...
Objectives
Long-term isoflurane anesthesia exposure could result in postoperative cognitive dysfunction (POCD). Preoperative stress is also reported to be a risk factor of POCD. However, it is unknown whether acute stress could impair memory after long-term isoflurane anesthesia.
Methods
In this study, we categorized the mice with acute stress int...
Antisense locked nucleic acid (LNA) technology has been widely used for silencing microRNAs with enhanced specificity and efficiency. In this protocol, we first describe the procedure for targeted intracranial delivery of LNAs to silence microRNAs specifically in the mouse brain. We then detail the steps to isolate RNA and protein from mouse brain,...
The DNA-PK enzyme phosphorylates the Ser15 and Ser37 residues in the TAD domain of p53. Mainly, the Ser15 phosphorylation event prevents p53 from interacting with MDM2 and stabilizes it. It has been reported that Phe19Arg and Leu22Gln-Trp23Ser mutations located close to Ser15 cause tumorigenesis in mouse models. These mutations are also known to re...
At this study positive cases of infected sheep by the parasite were identified, and distinguished from other infected cases caused by other Taenia species. This was done by applying ELISA technique for identifying positive cases and followed by Western-blot analysis for infection confirmation, which will help in correct treatment and prevent the sp...
Results presented on the "Venoms and Toxins 2022" conference in Oxford.
Western blot is a technique used to separate proteins by size followed by detection using antibodies specific to the protein of interest. This protocol describes the basic steps for lysing cells, determining total protein concentration in the lysate, running a precast SDS-PAGE gel, and immunoblotting.
Sarcocystis neurona é o principal agente causador da mieloencefalite protozoária equina. No Brasil, a soroprevalência pode chegar a mais de 90%, variando de acordo com o tamanho amostral e o teste sorológico utilizado. No presente estudo, soropositividade e titulação de anticorpos (IgG) contra S. neurona (SN138) e Sarcocystis sp. (Dal23-CG filogene...
This protocol describes how to perform a west blotting technique using fly heads.
Here, we describe the procedure by which human iPSC-derived neurons or mouse embryonic fibroblasts (MEFs) were lysed and probed for levels of proteins of interest using Western blot.
Background:
Systemic lupus erythematosus (SLE), especially neuropsychiatric SLE (NPSLE), is a complex systemic autoimmune disease, characterized by variable course and multiple organ dysfunction. Our study aimed to identify crucial microRNA (miRNAs) in SLE and NPSLE.
Methods:
Totally 12 cases of serum specimens were collected from General Hospit...
AMAÇ: Anjiyotensin II (Ang II)’nin damar duvarındaki asıl hedefi vasküler düz kas hücreleri (VDKH)’dir. Bu hücrelerin proliferasyonunu uyararak ateroskleroz ve hipertansiyon patogenezine katılır. Yüksek konsantrasyondaki glukoz (YG) da bu hücrelerde proliferasyonu artırarak diyabetlilerde görülen hızlandırılmış ateroskleroz sürecine katkıda bulunur...
A series of quillaic acid derivatives with different substituents on the 28-carboxyl group were designed and synthesized. Five human cancer cell lines (HCT116, BEL7402, HepG2, SW620, and MCF-7) were evaluated for their antitumor activity in vitro. Some of the tested derivatives showed improved antiproliferative activity compared to the lead compoun...
Introducción: los estudios de prevalencia de VIH en personas trans de Colombia son escasos, es necesario aumentar la investigación en este grupo para mejorar la planeación e implementación de programas de detección, prevención, control y tratamiento. Objetivo: determinar la prevalencia de VIH en personas trans atendidos en una institución prestador...
Li T, Wan Y, Su Z, Li J, Han M, Zhou C. Cancer Manag Res. 2020;12:6477–6491.
The Editor and Publisher of Cancer Management and Research wish to retract the published article. Concerns were raised regarding the alleged similarity in features of the western blots from Figure 8A and B with those from unrelated papers.
The authors responded to our quer...
Objective:
The aim of the study was to explore the relevance of CD40, CD86, and GSTO1 with the pathogenesis of COPD.
Methods:
Patients with acute exacerbation of COPD were contrasted with the healthy and nonsmoking ones and smoking but without COPD ones. The changes of CD40, CD86, and GSTO1 in the peripheral blood, collected from different group...
The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene encodes for a chloride channel defective in Cystic Fibrosis (CF). Accordingly, upregulation of its expression might be relevant for the development of therapeutic protocols for CF. MicroRNAs are deeply involved in the CFTR regulation and their targeting with miRNA inhibitors (inclu...
目的:制备可用于甘蔗花叶病毒（ScMV）E株系（ScMV-E）检测用多克隆抗体。方法:将ScMV-E外壳蛋白（CP）基因连接到pET29a（+）上,经PCR检测、酶切及测序鉴定获得重组质粒pET29a-CP,在大肠杆菌BL21（DE3）中诱导表达重组ScMV-E外壳蛋白;采用His Trap Kit纯化目的蛋白,作为抗原免疫新西兰大白兔,制备特异性抗体;通过间接ELISA、Western blot和组织印迹法检测所制备抗体的特异性。结果:SDS-PAGE分析表明,重组融合蛋白含6个组氨酸标记,相对分子质量约43000;Western blot检测显示所获得的抗体特异性良好,间接ELISA法测得血清的效价为1:81 920;甘蔗叶片的组织印迹检测结果显示杂交效果良好。结论:制备的多克隆抗体可直...
Here we present a general protocol for immunological detection by Western blotting MTOR, MTOR (pS2448), ULK1, ULK1 (pS757), p70S6K, p70S6K (pT389), SQSTM1, CALCOCO2, MAP1LC3B, GABARAP, TFEB, TFE3, PGRN, HSP90, and PCNA
This protocol describes the protein extraction from human alveolar-like organoids followed by western blotting. It gives a detailed description of the preparation of cell lysate from human alveolar-like organoids, followed by exact steps to perform a western blot and the immunostaining to detect ACE2, TMPRSS2, and FURIN as host factors of SARS-CoV-...
An interaction between mitochondrial dynamics, physical activity levels, and COVID-19 severity has been previously hypothesized. However, this has not been tested. We aimed to compare mitochondrial morphology and cristae den- sity of PBMCs between subjects with non-severe COVID-19, subjects with se- vere COVID-19, and healthy controls. Additionally...
Introducción:
La diabetes mellitus (DM) inhibe la biosíntesis de serotonina cerebral mediante cambios en la actividad y expresión de triptófano-5-hidroxilasa (TPH).
Objetivos:
Determinar si los cambios en la expresión de TPH1 y TPH2 cerebral y en el número de neuronas serotoninérgicas causados por la DM retornan a la normalidad en ratas con diabet...
This protocol details the procedure to assess mitophagy by analysing COXII degradation via Western blotting.
Background: Diet-induced obesity is associated with several cardiovascular diseases and myocardial apoptosis. This study aimed to investigate the effect of high intensity interval training (HIIT) on the myocardial apoptosis in the diet-induced obese rats. Methods: Thirty-two male rats, after an obesity induction period, were randomly selected and a...
Introducción: La lesión traumática de la médula espinal es la principal causa de discapacidad motora en el mundo, y representa una prioridad para la Organización Mundial de la Salud. Se estudió, a nivel estructural y bioquímico, el efecto de la hipotermia sobre la expresión de la CIRBP (proteína activada por frío) en el asta anterior de la médula d...
目的
探讨阻断p21蛋白激活激酶1（P21 Activated Kinase 1，PAK1）活性对急性巨核细胞白血病（AMKL）细胞株（CHRF和CMK）增殖、分化和细胞凋亡的影响。
方法
采用细胞计数法检测PAK1抑制剂（IPA-3和G5555）作用前后AMKL细胞增殖及集落形成能力；应用流式细胞术检测PAK1抑制剂对AMKL细胞周期的影响；Western blot法检测细胞周期蛋白cyclin D1和细胞凋亡相关蛋白Cleaved caspase 3的表达；使用慢病毒介导的shRNA转染技术干扰AMKL细胞中PAK1的蛋白表达水平，应用流式细胞术检测敲低PAK1激酶的活性对AMKL细胞中多倍体DNA形成能力和细胞凋亡的影响。
结果
PAK1抑制剂以剂量依赖性的方式抑制AMKL细胞的增...
The high sensitivity of the automated tests used for Toxoplasma gondii serology can yield false-positive IgM results due to aspecific reactions. On the other hand, specific therapy can delay IgG production and, therefore, the diagnosis of seroconversion. There is a need for confirmation tests to early detect seroconversions during pregnancy. We con...
The AK423 antibody, raised against Dictyostelium actin, recognizes both Dictyostelium and human actin by western blot. However, it recognizes Dictyostelium actin with higher affinity than human actin.
La equinococosis quística (EQ) es una zoonosis parasitaria causada por Echinococcus granulosus sensu lato. Técnicas inmunodiagnósticas como Western blot (WB) o el enzimoinmunoensayo (ELISA), con distintos antígenos, pueden ser aplicadas al diagnóstico de la EQ ovina con fin tales de vigilancia epidemiológica en programas de control. Sin embargo, ha...
Az emlődaganatok kezelése sokat fejlődött, a relapszusok és a rossz prognózisú esetek előfordulása azonban új terápiás megközelítéseket igényel. A primer és metasztatikus daganatsejtek fontos jellemzője a metabolikus folyamataik átrendeződése, ezt befolyásolják a genetikai, mikrokörnyezeti változások és a tumorszövet heterogenitás is. A bioenergeti...
AF397, AK652, AN193 and AV442 antibodies against the GFP protein recognize a GFP-tagged human TAC protein by western blot.
Although bortezomib (BTZ) displays efficacy in treating multiple myeloma (MM), BTZ resistance in MM patients has been reported. Meanwhile, treating BTZ resistant MM cells with β-catenin inhibitors have demonstrated the ability to reserve BTZ resistance. Thus, the present study aimed to investigate the synergistic effect of the β-catenin inhibitors,...
Pada penelitian awal didapatkan diet restriksi vitamin B12 menyebabkan hiperhomosisteinemia dan resistensi insulin, yang ditandai oleh hiperglikemia dan meningkatnya nilai Homeostatic Model Assessment for Insulin Resistance (HOMA-IR). Dengan menggunakan sampel jaringan biologik tersimpan, hati tikus Spraque-Dawley dari penelitian tersebut, peneliti...
The AK423 antibody, raised against Dictyostelium actin, recognizes human actin by western blot; AE765, AK692, AO233 and AO234 do not.
SDF2L1 is a new type of endoplasmic reticulum stress inducible protein, which is related to poor prognosis of various cancer, we initially studied the low expression level of SDF2L1 in NPC, but the molecular mechanism of SDF2L1 in NPC needs further elucidation. To identify phosphorylated proteins regulated by SDF2L1 in nasopharyngeal carcinoma (NPC...
The N-linked glycoprotein profile of GCase trapped in the ER makes it susceptible to digestion by the glycosidase EndoH. Western blotting of GCase following EndoH digestion will yield an additional lower molecular weight band that corresponds to the GCase that is trapped in the ER.
Hipótesis: La diabetes tipo 1 (DT1) es una enfermedad crónica caracterizada por un ataque autoinmune a las células β pancreáticas. En el transcurso de la enfermedad, las células α pancreáticas están expuestas al mismo ambiente proinflamatorio, pero mientras que éstas sobreviven, la masa de células β se pierde progresivamente. Entre los mecanismos m...
(1) Background: When red blood cells are centrifuged in a continuous Percoll-based density gradient, they form discrete bands. While this is a popular approach for red blood cell age separation, the mechanisms involved in banding were unknown. (2) Methods: Percoll centrifugations of red blood cells were performed under various experimental conditio...
Voltage-gated calcium channel (VGCC) subunits have been genetically associated with autism spectrum disorders (ASD). The properties of the pore-forming VGCC subunit are modulated by auxiliary β-subunits, which exist in four isoforms (Ca V β 1-4 ). Our previous findings suggested that activation of L-type VGCCs is a common feature of Ca V β 2 subuni...
The recombinant antibodies AI334, AQ806, AR222, AR249, AS274, AS702, AS708, RB590, RB591 and RB596 detect by western blot the spike S protein from SARS-CoV-2.
This is a protocol for detection of overexpressed and endogenous LPPR3 from N1E-115 cells and primary hippocampal neurons.
Here we present a general protocol for immunological detection by Western blotting of APP and proteins of the endolysosomal system, including EEA1, RAB5, PSEN1, LAMP1, LAMP2, TMEM192, and BACE1.
Since epsilon-toxin (ETX) is necessary for the virulence of C. perfringens type D and, likely, type B strains, understanding the regulation of ETX production is important. In 2011, we reported that an agrB null mutant of type D strain CN3718 produces less ETX than its wild-type parent.
This protocol describes the protein extraction from human alveolar-like organoids followed by western blotting. It gives a detailed description of the preparation of cell lysate from human alveolar-like organoids, followed by exact steps to perform a western blot and the immunostaining to detect ACE2, TMPRSS2, and FURIN as host factors of SARS-CoV-...
Introdução:Os kits empregados no diagnóstico do vírus da imunodeficiência humana (HIV) devem cumprir requisitos da RDC n° 36, de 26 de agosto de 2015, para registro junto à Agência Nacional de Vigilância Sanitária e da Lei n° 6.360, de 23 de setembro de 1976, para comercialização no país. Uma das etapas do registro é a análise prévia laboratorial d...
The recombinant antibodies AF394, AF395, AF397, AK142, AK652, AN193 and AV441 detect the GFP protein by western blot.
![The overall architecture of CSLN [1]. Get the molecular fingerprint...](publication/358344382/figure/fig1/AS:1119593221357573@1643943233132/The-overall-architecture-of-CSLN-1-Get-the-molecular-fingerprint-through-MACCSkeys_Q320.jpg)
Background
Chinese herbal medicine is made up of hundreds of natural drug molecules and has played a major role in traditional Chinese medicine (TCM) for several thousand years. Therefore, it is of great significance to study the target of natural drug molecules for exploring the mechanism of treating diseases with TCM. However, it is very difficul...
Duan Y, Zhao M, Jiang M, Li Z, Ni C. Onco Targets Ther. 2020;13:2701–2710.
At the authors request, the Editor and Publisher of OncoTargets and Therapy wish to retract the published article. The authors have informed the journal that they have concerns with the reliability of the published results. It was also found that several western blot images...