Tag: HEK293

Biochemical characterization of the two novel mgCas12a proteins from the human gut metagenome

Discovery of 11 new Cas12a candidates from a human gut microbiome database To discover new members of the Cas12a protein family, we built a pipeline that searches and sorts out new candidates from the National Center for Biotechnological Information (NCBI) human metagenome database. Using this search pipeline (Fig. 1a), we initially…

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Modulation of C3a Activity: Internalization of the Human C3a Receptor and its Inhibition by C5a1 | The Journal of Immunology

The C3a receptor (C3aR) is expressed on most human peripheral blood leukocytes with the exception of resting lymphocytes, implying a much higher pathophysiological relevance of the anaphylatoxin C3a as a proinflammatory mediator than previously thought. The response to this complement split product must be tightly regulated in situations with sustained…

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Ammonium transport properties of HEK293 cells expressing RhCG mutants: preliminary analysis of structure/function by site-directed mutagenesis.

Zidi-Yahiaoui N, Ripoche P, Le Van Kim C, Gane P, D’Ambrosio AM, Cartron JP, Colin Y, Mouro-Chanteloup I Ammonium transport properties of HEK293 cells expressing RhCG mutants: preliminary analysis of structure/function by site-directed mutagenesis. Transfus Clin Biol. 2006 Mar-Apr;13(1-2):128-31. Epub 2006 Apr 3. We have recently shown by monitoring intracellular…

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How To Ensure Batch-To-Batch Media Consistency Before Starting Your Bioprocess

RPMI media is one of the most used chemically-defined and protein-free cell media since it works across a variety of mammalian cell lines (e.g., stem cells, T and B cells, hybridomas, HEK293, THP-1 leukemia cells, etc.) grown in suspensions and as monolayer cultures. RPMI media is usually supplemented with fetal…

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Affinity microfluidics enables high-throughput protein degradation analysis in cell-free extracts

We designed pDOC to support multiple strategies for analyzing protein degradation in vitro, specifically cell-free systems. The device is based on a MITOMI integrated microfluidic chip, originally developed to quantify protein-ligand interactions at equilibrium24,33. The MITOMI module was modified to contain an array of 16 × 64, 32 × 32…

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6 weird food myths that just won’t die

I’ve been noticing a theme on food TikTok lately. There are, of course, hundreds of myths and misconceptions regarding food that are perpetuated all over social media every single day. That’s nothing new, but there are several that, despite being debunked time and time again, just won’t die. They’ve basically…

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Nanobody-based RFP-dependent Cre recombinase for selective anterograde tracing in RFP-expressing transgenic animals

Screening of efficient mCherry-binding protein (MBP) pairs to design Cre recombinase dependent on RFPs First, we aimed to construct Cre recombinase dependent on RFPs based on the reported tool named Cre-DOG30. In this system, N-terminal and C-terminal split Cre recombinase fragments are fused with specific nanobodies for target proteins, and…

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HEK293-Transfection Reagent (BT-000015)

BOC Sciences HEK293-Transfection Reagent is a unique formulation based on polycationic liposomes of non-animal origin. It is designed for the transfection of HEK293 cells grown in suspension culture and is ideal for mammalian protein production. HEK293 cell line was first discovered in 1977, derived from human embryonic kidney cells transformed…

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Causes of ocular inflammation linked to AAV gene therapy

Low-grade inflammation seems to be associated with baseline disease in inherited retinal dystrophies (IRDs), according to Christine N. Kay, MD, a surgeon at Vitreoretinal Associates, in Gainesville, Florida. Examples of XX include zonular instability in cataract surgeries resulting in intraocular lens dislocations in retinitis pigmentosa (RP), presence of anterior vitreous…

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Transcription-independent regulation of STING activation and innate immune responses by IRF8 in monocytes

Reagents, antibodies, viruses and cells LMW-Poly(I:C), LPS, 2′3′-cGAMP and DMXAA (InvivoGen); hydroxyurea, camptothecin and mitomycin C (MCE); GM-CSF, Flt3L (peproTech); lipofectamine 2000 (Invitrogen); polybrene (Millipore); RNAiso Plus (Takara); HT-DNA (Sigma); SYBR (BIO-RAD); dual-specific luciferase assay kit (Promega); ELISA kit for murine Ifn-β (PBL); ELISA kits for murine IP-10 (Biolegend) were…

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Targeted inhibition of ubiquitin signaling reverses metabolic reprogramming and suppresses glioblastoma growth

Cell culture Human glioblastoma cells (U87MG and U87MG-Luc) and human embryonic kidney cells (HEK293) were obtained from the American Type Culture Collection (Manassas, Va.). Cells were cultured in Dulbecco’s Modified Eagle Medium supplemented with 10% fetal bovine serum (Gibco™ Fetal Bovine Serum South America, Thermo Scientific Fisher-US), 2 mM l-glutamine, 50 U/ml…

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Researchers Discover Novel Light-Gated Potassium Channel in Neurons

Summary: Researchers report they have identified the first natural light-gated potassium channel-rhodopsins. Source: Baylor College of Medicine A key approach to understanding the brain is to observe the behavioral effects of turning on specific populations of neurons. One of the most popular approaches to controlling neuronal activity in model systems…

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Transitioning from Quantitative PCR to Droplet Digital PCR for HEK293 Host Cell DNA Residual Testing

HEK293 cells are a dominant platform for cell and gene therapy production because they are highly effective at producing large amounts of recombinant viral vectors. However, it is critical to remove trace host cell contamination from therapeutic products to avoid endangering patients. To meet rigorous regulatory guidelines, biopharmaceutical manufacturers need…

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SARS-CoV-2 ORF8 protein binds to dendritic cells and induces a hyper-inflammatory cytokine storm

In a recent study posted to the bioRxiv* pre-print server, researchers examined how the interaction of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) open reading frame 8 (ORF8) protein with host dendritic cells (DCs) induced cytokine storm. ​​​​​​​Study: The unique ORF8 protein from SARS-CoV-2 binds to human dendritic cells…

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NP1 mutants that do not interact with Ku70 and RPA70 in vitro lose the function in facilitating HBoV1 DNA replication in HEK293 cells.

(A) Schematic digram of the NP1 protein. NP1 protein is schematically diagramed with predicated helical strutrues. The amino acid sequences of aa90-219 are shown with indication of each helix. The potential functional motifs (underlined) within each helical struture were predicted with Eukaryotic Linear Motif resource. NP1 mutants that bear mutaions…

Continue Reading NP1 mutants that do not interact with Ku70 and RPA70 in vitro lose the function in facilitating HBoV1 DNA replication in HEK293 cells.

Bispecific antibodies | Scientist Live

Bispecific antibodies: rising stars in antibody therapeutics Monoclonal antibodies (mAbs) are important therapeutic agents for the treatment of many human diseases, such as cancer, autoimmune diseases, cardiovascular diseases, asthma, and viral infections. Unlike monospecific mAbs, bispecific antibodies (bsAbs) are antibodies containing two antigen-binding sites and therefore can simultaneously target two…

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Recombinant Feline EGF Protein, HEK293 Cells, hFc Tag, 62010-U01H

EGF Protein, Feline, Recombinant (ECD, hFc Tag): 产品信息 纯度 > 90 % as determined by SDS-PAGE. 生物活性 Testing in progress 蛋白构建 A DNA sequence encoding the Feline EGF (NP_001009381.1) (Asn969-Arg1020) was expressed with the fused Fc region of human IgG1 at the N-terminus. 表达宿主 HEK293 Cells 分子量 The…

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Researchers identify cellular serpins as host factors that restrict SARS-CoV-2 infection

The mass vaccination programs against coronavirus disease 2019 (COVID-19) received massive media attention and were widely credited for reducing the threat of the disease. However, many countries struggle to obtain enough doses to immunize their population, and some immunocompromised individuals are less protected by the vaccines. There is still a…

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Recent developments in miRNA based recombinant protein expression in CHO

Aguiar TQ, Santos SB, Martins IM, Domingues L, Oliveira C (2019) Production and bioengineering of recombinant pharmaceuticals. Proteins: sustainable source, processing and applications. Elsevier, Amsterdam, pp 259–293 Chapter  Google Scholar  Amadi IM, Agrawal V, Christianson T, Bardliving C, Shamlou P, LeBowitz JH (2020) Inhibition of endogenous miR-23a/miR-377 in CHO cells…

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Recombinant Human ACE2 Protein, HEK293 Cells, hFc Tag, 10108-H02H-B

ACE2 Protein, Human, Recombinant (hFc Tag), Biotinylated: Product Information Purity > 95 % as determined by SDS-PAGE. Activity Testing in progress Protein Construction A DNA sequence encoding the extracellular domain (Met 1-Ser 740) of human ACE2 precursor (NP_068576.1) was expressed with the fused Fc region of human IgG1…

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Full Text Journal Article: Androgen receptor transcriptional activity and chromatin modifications on the ABCB1/MDR gene are critical for taxol resistance in ovarian cancer cells.

PDF.MPro – Full Text Journal Article: Androgen receptor transcriptional activity and chromatin modifications on the ABCB1/MDR gene are critical for taxol resistance in ovarian cancer cells. Home / Search / Abstract 30317630 (Page 1) Advertisement Find full text journal articles PMID: 30317630 (view PubMed database entry)DOI: 10.1002/jcp.27535 (read at publisher’s…

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Study explores SARS-CoV-2 Omicron variant and infected colon epithelial cells

In a recent study posted to the bioRxiv* preprint server, researchers assessed the impact of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variant on infected human cells and organs. Study: SARS-CoV-2 Omicron BA.1 variant infection of human colon epithelial cells. Image Credit: PHOTOCREO Michal Bednarek/Shutterstock Various studies have…

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Recombinant Influenza B Hemagglutinin/HA Protein, HEK293 Cells, His Tag, 40862-V08H

The influenza viral Hemagglutinin (HA) protein is a homotrimer with a receptor binding pocket on the globular head of each monomer.HA has at least 18 different antigens. These subtypes are named H1 through H18.HA has two functions. Firstly, it allows the recognition of target vertebrate cells, accomplished through the…

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Does Norepinephrine Metabolite DOPEGAL Turn Tau Toxic?

08 Apr 2022 What makes tau toxic? In the March 24 Nature Structural & Molecular Biology, researchers led by Keqiang Ye at Shenzhen Institute of Advanced Technology, China, suggest a new culprit, a norepinephrine metabolite known as DOPEGAL, which is produced only in the locus coeruleus (LC). The authors found…

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Human GITR HEK293 Cells Overexpression Lysate, 13643-H08HL

制备方法 Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min…

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The Ribosomal Protein RpL22 Interacts In Vitro with 5′-UTR Sequences Found in Some Drosophila melanogaster Transposons

Mobility of eukaryotic transposable elements (TEs) are finely regulated to avoid an excessive mutational load caused by their movement. The transposition of retrotransposons is usually regulated through the interaction of host- and TE-encoded proteins, with non-coding regions (LTR and 5′-UTR) of the transposon. Examples of new potent cis-acting sequences, identified…

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Transcriptional kinetics and molecular functions of long noncoding RNAs

Ethical compliance The research carried out in this study has been approved by the Swedish Board of Agriculture, Jordbruksverket: N343/12. Cell culture Mouse primary fibroblasts were derived from adult (>10 weeks old) CAST/EiJ × C57BL/6J or C57BL/6J × CAST/EiJ mice by skinning, mincing and culturing tail explants (for at least 10 d) in DMEM high…

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Study on the Clinical and Functional Characterization of Ryanodine Receptor 2 Variants Implicated in Calcium-Release Deficiency Syndrome

To enhance CRDS insight through international cooperation. In the multicenter observational trial, individuals with unexplained life-threatening arrhythmic events and an ultrarare RyR2 mutation were identified. The effects of excitotoxicity-related variants were investigated in HEK293 cells treated with caffeine to determine their functional impact. The researchers reviewed data from September 1,…

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Hypoxia-induced HIF1alpha activation regulates small extracellular vesicle release in human embryonic kidney cells

This article was originally published here Sci Rep. 2022 Jan 27;12(1):1443. doi: 10.1038/s41598-022-05161-7. ABSTRACT Extracellular vesicles (EVs) are membrane enclosures released by eukaryotic cells that carry bioactive molecules and serve to modulate biological responses in recipient cells. Both increased EV release and altered EV composition are associated with the development…

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The Fabrication and Operation of a Continuous Flow, Micro-Electroporation System with Permeabilization Detection

Current therapeutic innovations, such as CAR-T cell therapy, are heavily reliant on viral-mediated gene delivery. Although efficient, this technique is accompanied by high manufacturing costs, which has brought about an interest in using alternative methods for gene delivery. Electroporation is an electro-physical, non-viral approach for the intracellular delivery of genes…

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A cell-based phenotypic library selection and screening approach for the de novo discovery of novel functional chimeric antigen receptors

Cell lines and culture Unless stated otherwise, all tumor target cell lines were sourced from the ATCC via LGC Standards. Human endogenous mesothelin (MSLN)-positive target cell lines H-226 (lung carcinoma, MSLN+, ATCC® CRL-5826™) and AsPC-1 (ATCC CRL-1682), and MSLN-negative Raji control cells (ATCC CCL-86) were cultured in RPMI-1640 Glutamax (Life Technologies,…

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Recombinant Human CD94/KLRD1 Protein, HEK293 Cells, His Tag, 12649-H07H

CD94/KLRD1 Protein, Human, Recombinant (His Tag): 产品信息 纯度 > 95 % as determined by SDS-PAGE 生物活性 Testing in progress 蛋白构建 A DNA sequence encoding the human KLRD1 (Q13241-1) (Lys32-Ile179) was expressed with a polyhistidine tag at the N-terminus. 表达宿主 HEK293 Cells 分子量 The recombinant human KLRD1 consists of…

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A novel antagonist of TRPM2 and TRPV4 channels: Carvacrol

This article was originally published here Metab Brain Dis. 2022 Jan 6. doi: 10.1007/s11011-021-00887-1. Online ahead of print. ABSTRACT The overload cytosolic free Ca2+ (cCa2+) influx-mediated excessive generation of oxidative stress in the pathophysiological conditions induces neuronal and cellular injury via the activation of cation channels. TRPM2 and TRPV4 channels…

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Addressing the Challenges of AAV Production

Adeno-associated virus (AAV) vectors are a versatile and appealing gene therapy delivery platform, capable of targeting a wide range of cell types. However, AAV must be produced in a living system, and issues with scalability and high production costs have so far limited the widespread adoption of the technology. Technology…

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Sod1 integrates oxygen availability to redox regulate NADPH production and the thiol redoxome

Significance Cu/Zn superoxide dismutase (Sod1) is a key antioxidant enzyme, and its importance is underscored by the fact that its ablation in cell and animal models results in oxidative stress; metabolic defects; and reductions in cell proliferation, viability, and lifespan. Curiously, Sod1 detoxifies superoxide radicals (O2•−) in a manner that…

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Figure 3 | BK Polyomavirus Activates HSF1 Stimulating Human Kidney Hek293 Cell Proliferation

BKPyV induces Hek293 cell proliferation at late time of infection. Hek293 cells were infected with BKPyV for 1 h. (a, b) Cells were counted by Trypan blue exclusion after 24 h. Data are expressed as (; ; ). (c) Cell proliferation was determined through immunofluorescence detection of incorporated BrdU. Data are expressed…

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NPSR1 / NPSR / GPR154 Protein Over-Expression Lysate

Western blot Western validation with an anti-DDK antibody * L: Control HEK293 lysate R: Over-expression lysate Western blot Western validation with an anti-DDK antibody * L: Control HEK293 lysate R: Over-expression lysate Western blot Western validation with an anti-DDK antibody * L: Control HEK293 lysate R: Over-expression…

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Single-cell delineation of lineage and genetic identity in the mouse brain

STICR lentiviral library preparation and validation We synthesized a high-complexity lentivirus barcode library that encodes approximately 60–70 million distinct oligonucleotide RNA sequences (STICR barcodes). STICR barcodes comprised three distinct oligonucleotide fragments cloned sequentially into a multicloning site within the 3′ UTR of an enhanced green fluorescent protein (eGFP) transgene under…

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WT1 regulates HOXB9 gene expression in a bidirectional way

This article was originally published here Biochim Biophys Acta Gene Regul Mech. 2021 Sep 8:194764. doi: 10.1016/j.bbagrm.2021.194764. Online ahead of print. ABSTRACT The homeoboxB9 (HOXB9) gene is necessary for specification of the anterior-posterior body axis during embryonic development and expressed in various types of cancer. Here we show that the…

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CRISPR Applications in Medicine Depend on Minimizing Off-Target Editing

By Rolf Turk, PhD Genome editing technologies, such as CRISPR, are being applied to better understand basic biological systems as well as to research new kinds of gene and cell therapies. The CRISPR-Cas9 system comprises a Cas9 endonuclease protein that forms a complex with a guide RNA (gRNA) molecule, which…

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