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Clone Cells

Clone cells refer to a population of genetically identical cells derived from a single common ancestor cell.
These cells are widely used in biomedical research, including stem cell studies, cancer research, and genetic engineering.
Clone cells provide a valuable model system for investigating cellular processes, gene expression, and therapeutic development.
Researchers can utilize advanced AI-powered tools, such as PubCompare.ai, to optimize their clone cell experiments by easily locating and comparing the best protocols from literature, preprints, and patents.
These tools enhance reproducibility and streamline the research process, driving forward the field of clone cell biology.

Most cited protocols related to «Clone Cells»

1
Robust Knock-in Mouse Line Generation
Targeting constructs were generated using a combined gene synthesis (GenScript Corp.) and molecular cloning approach. Briefly, to target the Rosa26 locus, a cassette containing the following components was constructed: FRT – LoxP – Stop codons – 3x SV40 polyA – LoxP – EYFP – WPRE – bGH polyA – AttB – PGK promoter – FRT – Neo – PGK polyA – AttP. For most targeting vectors, this cassette was cloned into a Rosa-CAG targeting vector3 (link), downstream of the CAG promoter and upstream of the 3′ arm, to generate the final EYFP targeting vector. Unique restriction sites flanking the EYFP gene were used to replace EYFP with alternative reporter genes. For the Ai2 vector, which lacks the WPRE, the CAG promoter was inserted between the first FRT and LoxP sites, and the cassette was cloned immediately downstream of the 5′ homology arm. The final targeting vectors contained 5′ and 3′ homology arms of 1.1 kb and 4.3 kb, as well as a PGK-DTA cassette for negative selection. Targeting constructs for knock-in Cre lines inserted into other gene loci were constructed in similar ways.
The targeting vectors were linearized and transfected into the 129/B6 F1 hybrid ES cell line G442 (link) using an Amaxa electroporator. G418-resistant ES clones were screened by Southern blot analysis of HindIII digested DNA, which was probed with a 1.1 kb genomic fragment from immediately upstream of the 5′ arm. We observed a recombination rate of about 25% for the four constructs. Positive ES clones were injected into C57BL/6J blastocysts to obtain chimeric mice following standard procedures. Both ES cell transfections and blastocyst injections were performed by the University of Washington Transgenic Resources Program. Due to the robustness of the G4 cells, high-percentage chimeras and high rates of germline transmission were routinely obtained. Chimeric mice were bred with either C57BL/6J mice to obtain germline transmission or various Cre-driver lines for direct characterization.
An Ai9 ES cell clone with strong germline transmission potency was used in subsequent transfections for the Flp-mediated exchange strategy outlined in Supplementary Figure 4 online. Ai9 ES cells were co-transfected using a Bio-Rad electroporator with 100 μg of pCAGGS-FLPe (Open Biosystems) and 40 μg of an incoming replacement vector. After 8 to 10 days of Hygromycin B selection, surviving colonies that also appeared green by fluorescence microscopy were picked and screened by PCR using primer sets designed to confirm a correct insertion of the incoming vector at the 5′ and 3′ FRT recombinase sites.
Madisen L., Zwingman T.A., Sunkin S.M., Oh S.W., Zariwala H.A., Gu H., Ng L.L., Palmiter R.D., Hawrylycz M.J., Jones A.R., Lein E.S, & Zeng H. (2009). A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nature neuroscience, 13(1), 133-140.
Publication 2009
Anabolism Animals, Transgenic antibiotic G 418 Blastocyst Cells Chimera Clone Cells Cloning Vectors Codon, Terminator Embryonic Stem Cells Genes Genes, Reporter Genetic Loci Genome Germ Cells Germ Line Hybrid Cells Hygromycin B Mice, Inbred C57BL Microscopy, Fluorescence Mus N-fluoresceinylphosphatidylethanolamine Oligonucleotide Primers Poly A Recombinase Recombination, Genetic Rosa Simian virus 40 Southern Blotting Transfection Transmission, Communicable Disease
2
Robust Knock-in Mouse Line Generation
Targeting constructs were generated using a combined gene synthesis (GenScript Corp.) and molecular cloning approach. Briefly, to target the Rosa26 locus, a cassette containing the following components was constructed: FRT – LoxP – Stop codons – 3x SV40 polyA – LoxP – EYFP – WPRE – bGH polyA – AttB – PGK promoter – FRT – Neo – PGK polyA – AttP. For most targeting vectors, this cassette was cloned into a Rosa-CAG targeting vector3 (link), downstream of the CAG promoter and upstream of the 3′ arm, to generate the final EYFP targeting vector. Unique restriction sites flanking the EYFP gene were used to replace EYFP with alternative reporter genes. For the Ai2 vector, which lacks the WPRE, the CAG promoter was inserted between the first FRT and LoxP sites, and the cassette was cloned immediately downstream of the 5′ homology arm. The final targeting vectors contained 5′ and 3′ homology arms of 1.1 kb and 4.3 kb, as well as a PGK-DTA cassette for negative selection. Targeting constructs for knock-in Cre lines inserted into other gene loci were constructed in similar ways.
The targeting vectors were linearized and transfected into the 129/B6 F1 hybrid ES cell line G442 (link) using an Amaxa electroporator. G418-resistant ES clones were screened by Southern blot analysis of HindIII digested DNA, which was probed with a 1.1 kb genomic fragment from immediately upstream of the 5′ arm. We observed a recombination rate of about 25% for the four constructs. Positive ES clones were injected into C57BL/6J blastocysts to obtain chimeric mice following standard procedures. Both ES cell transfections and blastocyst injections were performed by the University of Washington Transgenic Resources Program. Due to the robustness of the G4 cells, high-percentage chimeras and high rates of germline transmission were routinely obtained. Chimeric mice were bred with either C57BL/6J mice to obtain germline transmission or various Cre-driver lines for direct characterization.
An Ai9 ES cell clone with strong germline transmission potency was used in subsequent transfections for the Flp-mediated exchange strategy outlined in Supplementary Figure 4 online. Ai9 ES cells were co-transfected using a Bio-Rad electroporator with 100 μg of pCAGGS-FLPe (Open Biosystems) and 40 μg of an incoming replacement vector. After 8 to 10 days of Hygromycin B selection, surviving colonies that also appeared green by fluorescence microscopy were picked and screened by PCR using primer sets designed to confirm a correct insertion of the incoming vector at the 5′ and 3′ FRT recombinase sites.
Madisen L., Zwingman T.A., Sunkin S.M., Oh S.W., Zariwala H.A., Gu H., Ng L.L., Palmiter R.D., Hawrylycz M.J., Jones A.R., Lein E.S, & Zeng H. (2009). A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nature neuroscience, 13(1), 133-140.
Publication 2009
Anabolism Animals, Transgenic antibiotic G 418 Blastocyst Cells Chimera Clone Cells Cloning Vectors Codon, Terminator Embryonic Stem Cells Genes Genes, Reporter Genetic Loci Genome Germ Cells Germ Line Hybrid Cells Hygromycin B Mice, Inbred C57BL Microscopy, Fluorescence Mus N-fluoresceinylphosphatidylethanolamine Oligonucleotide Primers Poly A Recombinase Recombination, Genetic Rosa Simian virus 40 Southern Blotting Transfection Transmission, Communicable Disease
3
Analyzing Genotypic Diversity Using Poppr
Once data is imported into R, the user can dynamically access and manipulate the population hierarchy with the function splitcombine(), subset the data set by population with popsub(), and check for cloned multilocus genotypes using mlg(). For data sets that include clones, the poppr function clonecorrect() will censor clones with respect to any level of a population hierarchy. In the case of missing data we use the commonly implemented, most parsimonious approach of treating missing states as novel alleles. This inherently makes analysis sensitive to missing data and genotyping error, but the user has tools available such as missingno() to filter out missing data at a per-individual or per-locus level. The user can also decide how uninformative loci (e.g., alleles occurring at minor frequencies; monomorphic loci; fixed heterozygous loci) are treated using the function informloci(). Thus, the user can specify a frequency for removal of uninformative loci. The user is encouraged to conduct analysis with and without missing data/uninformative loci to assess sensitivity to these issues when making inferences. A full list of functions available in poppr is provided in Table 1.
Typical analyses in poppr start with summary statistics for diversity, rarefaction, evenness, MLG counts, and calculation of distance measures such as Bruvo’s distance, providing a suitable stepwise mutation model appropriate for microsatellite markers (Bruvo et al., 2004 (link)). Poppr will define MLGs in your data set, show where they cross populations, and can produce graphs and tables of MLGs by population that can be used for further analysis with the R package vegan (Oksanen et al., 2013 ). Many of the diversity indices calculated by the vegan function diversity() are useful in analyzing the diversity of partially clonal populations. For this reason, poppr features a quick summary table (Table 2) that incorporates these indices along with the index of association, IA (Brown, Feldman & Nevo, 1980 (link); Smith et al., 1993 (link)), and its standardized form, r¯d , which accounts for the number of loci sampled (Agapow & Burt, 2001 (link)). Both measures of association can detect signatures of multilocus linkage and values significantly departing from the null model of no linkage among markers are detected via permutation analysis utilizing one of four algorithms described in Table 3 (Agapow & Burt, 2001 (link)). The user can specify the number of samples taken from the observed data set to obtain the null distribution expected for a randomly mating population. Detailed examples of these analyses can be found in the poppr manual.
Kamvar Z.N., Tabima J.F, & Grünwald N.J. (2014). Poppr: an R package for genetic analysis of populations with clonal, partially clonal, and/or sexual reproduction. PeerJ, 2, e281.
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Publication 2014
Alleles Clone Cells Heterozygote Hypersensitivity Mutation Short Tandem Repeat Vegan
4
Comprehensive Manual Gene Annotation Workflow
The GENCODE gene set is created by merging the results of manual and computational gene annotation methods. Manual gene annotation has two major modes of operation: clone-by-clone and targeted annotation. ‘Clone-by-clone’ annotation involves ‘walking’ across a genomic region, investigating the sequence, aligned expression data and computational predictions for each BAC clone. In doing so, an expert annotator investigates all possible genic features and considers all possible annotations and biotypes simultaneously. We believe this approach carries substantial advantages. For example, the decision to annotate a locus as protein-coding or pseudogenic benefits from being able to weigh both possibilities in light of all available evidence. This process helps prevent false positive and false negative misclassifications. Targeted annotation is designed to answer specific questions such as ‘is there an unannotated protein-coding gene in this position?’ Ranked target lists are generated by computational analysis based, for example, on transcriptomic data, shotgun proteomic data or conservation measures. Over the last two years mouse annotation has been dominated by the clone-by-clone approach while the human genome has been refined entirely via targeted reannotation except for the annotation of human assembly patches and haplotypes released by the Genome Reference Consortium (15 (link)), which take a clone-by-clone approach.
Over the last two years, we have focused on two broad areas: completing the first pass manual annotation across the entire mouse reference genome and a dedicated effort to improve the annotation of protein-coding genes in human and mouse.
We have completed the annotation of novel protein-coding genes, lncRNAs and pseudogenes, plus QC and updating previous annotation where necessary for mouse chromosomes 9, 10, 11, 12, 13, 14, 15, 16 and 17. These updates bring the fraction of the mouse genome with completed first pass manual annotation to approximately 97%. In addition, we have continued to work with the NCBI and Mouse Genome Informatics project at the Jackson Laboratory to resolve annotation differences for protein-coding, pseudogene and lncRNA loci. For protein-coding genes this is under the umbrella of the Consensus Coding Sequence (CCDS) project (16 (link)).
We have also manually investigated unannotated regions of high protein-coding potential identified by whole genome analysis using PhyloCSF (17 (link)) (a tool described in more detail below). In human, this led to the addition of 144 novel protein-coding genes and 271 pseudogenes (of which 42 were unitary pseudogenes). In mouse, we annotated orthologous loci for all but 11 of the 144 human protein-coding genes. We have also revisited the annotation of all olfactory receptor loci in both human and mouse, using RNAseq data to define 5′ and 3′ UTR sequences for ∼1400 loci. In human we have also targeted a ‘deep dive’ manual reannotation of genes on clinical panels for paediatric neurological disorders to identify missing functional alternative splicing. Incorporating second and third generation transcriptomic data, we reannotated ∼190 genes and added more than 3600 alternatively spliced transcripts, including ∼1400 entirely novel exons and an additional ∼30kb of CDS. We have also completed an effort to capture all recently described unannotated microexons (18 (link)) into GENCODE, and further added an additional 146 novel microexons mined from public SLRseq data (19 (link)).
As part of the CCDS collaboration with RefSeq, we have checked a large subset of human loci where there was disagreement over gene biotype. Similarly, we have checked all UniProt manually annotated and reviewed (i.e. Swiss-Prot) accessions that lack an equivalent in GENCODE. As a result, we added 32 novel protein-coding loci to GENCODE and rejected more than 200 putative coding loci. Finally, we are manually reviewing genes previously annotated as protein-coding, but with weak or no support based on a method incorporating UniProt, APPRIS, PhyloCSF, Ensembl comparative genomics, RNA-seq, mass spectrometry and variation data (20 (link),21 (link)). Of the 821 loci investigated to date, 54 have had their coding status removed while a further 110 potentially dubious cases remain under review.
The approach taken reflects in the kinds of updates captured in the annotation. For example, the targeted reannotation in human leads to the annotation of few novel protein-coding loci but many novel transcripts at updated protein-coding and lncRNA loci. Conversely, in mouse the emphasis on clone-by-clone annotation identifies many more novel loci and transcripts across a broader range of biotypes (Figure 1).
Frankish A., Diekhans M., Ferreira A.M., Johnson R., Jungreis I., Loveland J., Mudge J.M., Sisu C., Wright J., Armstrong J., Barnes I., Berry A., Bignell A., Carbonell Sala S., Chrast J., Cunningham F., Di Domenico T., Donaldson S., Fiddes I.T., García Girón C., Gonzalez J.M., Grego T., Hardy M., Hourlier T., Hunt T., Izuogu O.G., Lagarde J., Martin F.J., Martínez L., Mohanan S., Muir P., Navarro F.C., Parker A., Pei B., Pozo F., Ruffier M., Schmitt B.M., Stapleton E., Suner M.M., Sycheva I., Uszczynska-Ratajczak B., Xu J., Yates A., Zerbino D., Zhang Y., Aken B., Choudhary J.S., Gerstein M., Guigó R., Hubbard T.J., Kellis M., Paten B., Reymond A., Tress M.L, & Flicek P. (2018). GENCODE reference annotation for the human and mouse genomes. Nucleic Acids Research, 47(Database issue), D766-D773.
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Publication 2018
3' Untranslated Regions Chromosomes, Human, Pair 9 Clone Cells Consensus Sequence Debility Exons Gene Annotation Gene Expression Profiling Gene Products, Protein Genes Genes, vif Genome Genome, Human Haplotypes Homo sapiens Mass Spectrometry Mice, Laboratory Nervous System Disorder NR4A2 protein, human Open Reading Frames Protein Annotation Proteins Pseudogenes Receptors, Odorant RNA, Long Untranslated RNA-Seq Staphylococcal Protein A TNFSF14 protein, human
5
Sequencing and Assembly of Rice Genome
The IRGSP clone and PCR sequences of the O. sativa (japonica group, cultivar Nipponbare) genome deposited in the International Nucleotide Sequence Databases as of 25 February 2010 were used in construction of the MTP. In addition, sequence reads generated by the Syngenta rice genome sequencing project (Goff et al. 2002 (link)) were assembled and used to extend contigs.
For the next-generation DNA sequencing of an NIAS individual, total genomic DNA was prepared from nuclei isolated from Nipponbare rice young leaves (two weeks after germination) using the CTAB method (Murray and Thompson 1980 (link)). The DNA samples were fragmented by a nebulizer or Branson Sonifier 250 (Danbury, CT). Sequencing libraries were constructed following the protocols with Illumina Genomic DNA Sample Preparation Kit and Roche GS DNA Library Preparation Kit, respectively. Illumina genome sequencing was performed by Illumina Genome Analyzer II/IIx with the Illumina version 2 sequencing kit. GS-FLX genome sequencing was performed using the Roche GS LR70 Sequencing Kit. The sequence reads are available at the DDBJ Sequence Read Archive (DRA000651).
For the CSHL individual, ~5 μg of Nipponbare rice genomic DNA was used as input for standard Illumina libraries. The DNA was sheared by adaptive focused acoustics using the Covaris (Woburn, MA) instrument and end-repaired using T4 DNA polymerase, Klenow fragment, and T4 polynucleotide kinase. Fragments were then treated with Klenow fragment (3’ - 5’ exonuclease) to add a single 3’ deoxyA overhang and ligated to standard paired-end Illumina adapters. Qiagen (Valencia, CA) columns were used for purification between steps. The fragments were size-selected at ~225 bp (including adapters) using agarose gel electrophoresis. The actual insert size excluding adapters was ~150 bp. The library was then PCR amplified using Phusion DNA polymerase in HF buffer for 14 cycles and quantified using the Agilent BioAnalyzer (Santa Clara, CA). All libraries were normalized to 10 nM before loading on the Illumina sequencers. Production sequencing was performed using Illumina GAIIx instruments with paired-end modules using the Illumina version 3 sequencing kits. The library was sequenced with 76 bp paired-end read lengths. Sequence data was processed using the Illumina GAPipeline v1.1 and v1.3.2 (Firecrest/Bustard v1.9.6 and Firecrest/Bustard v1.3.2). The sequence reads are available at the Sequence Read Archive of NCBI (SRX032913).
Syngenta rice genome sequences (Goff et al. 2002 (link)) were filtered by using IRGSP rice genomic sequences with similarity searches. The filtered sequences were then assembled; 50 large Syngenta contigs (between 4 kb and 40 kb), a total of 748 kb were used for potential gap filling.
Kawahara Y., de la Bastide M., Hamilton J.P., Kanamori H., McCombie W.R., Ouyang S., Schwartz D.C., Tanaka T., Wu J., Zhou S., Childs K.L., Davidson R.M., Lin H., Quesada-Ocampo L., Vaillancourt B., Sakai H., Lee S.S., Kim J., Numa H., Itoh T., Buell C.R, & Matsumoto T. (2013). Improvement of the Oryza sativa Nipponbare reference genome using next generation sequence and optical map data. Rice, 6, 4.
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Publication 2013
3'-5'-Exonucleases A-748 Acclimatization Acoustics Buffers Cell Nucleus Cetrimonium Bromide Clone Cells DNA-Directed DNA Polymerase DNA Library DNA Polymerase I Electrophoresis, Agar Gel Genome Germination Nebulizers Oryza sativa Polynucleotide 5'-Hydroxyl-Kinase

Most recents protocols related to «Clone Cells»

1
Quantifying IL-2 Effects on T Cell Activation
Not available on PMC !

Example 7

Impact of IL-2 signalling on Teff responses is characterised in a T cell activation assay, in which intracellular granzyme B (GrB) upregulation and proliferation are examined. Previously frozen primary human Pan T cells (Stemcell Technologies) are labelled with eFluor450 cell proliferation dye (Invitrogen) according to manufacturer's recommendation, and added to 96-U-bottom well plates at 1×105 cells/well in RPMI 1640 (Life Technologies) containing 10% FBS (Sigma), 2 mM L-Glutamine (Life Technologies) and 10,000 U/ml Pen-Strep (Sigma). The cells are then treated with 10 μg/ml anti-CD25 antibodies or control antibodies followed by Human T-Activator CD3/CD28 (20:1 cell to bead ratio; Gibco) and incubated for 72 hrs in a 37° C., 5% CO2 humidified incubator. To assess T cell activation, cells are stained with the eBioscience Fixable Viability Dye efluor780 (Invitrogen), followed by fluorochrome labelled antibodies for surface T cell markers (CD3-PerCP-Cy5.5 clone UCHT1 Biolegend, CD4-BV510 clone SK3 BD Bioscience, CD8-Alexa Fluor 700 clone RPA-T8 Invitrogen, CD45RA-PE-Cy7 clone HI100 Invitrogen, CD25-BUV737 clone 2A3 BD Bioscience) and then fixed and permeabilized with the eBioscience™ Foxp3/Transcription Factor Staining Buffer Set (Invitrogen) before staining for intracellular GrB and intranuclear FoxP3 (Granzyme B-PE clone GB11 BD Bioscience, FoxP3-APC clone 236A/E7). Samples are acquired on the Fortessa LSR X20 Flow Cytometer (BD Bioscience) and analysed using the BD FACSDIVA software. Doublets are excluded using FCS-H versus FCS-A, and lymphocytes defined using SSC-A versus FCS-A parameters. CD4+ and CD8+ T cell subsets gated from the live CD3+ lymphocytes are assessed using a GrB-PE-A versus proliferation eFluor450-A plot. Results are presented as percentage of proliferating GrB positive cells from the whole CD4+ T cell population. Graphs and statistical analysis is performed using GraphPad Prism v7. (results not shown)

US11873341B2. Anti-CD25 for tumour specific cell depletion (2024-01-16). Tusk Therapeutics Ltd. [GB], Cancer Research Technology Limited [GB]. Inventors: Anne Goubier [GB], Beatriz Goyenechea Corzo [GB], Josephine Salimu [GB], Kevin Moulder [GB], Pascal Merchiers [GB], Sergio Quezada [GB].
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Patent 2024
Anti-Antibodies Antibodies Biological Assay Buffers CD4 Positive T Lymphocytes Cell Proliferation Cells Clone Cells CY5.5 cyanine dye Eragrostis Fluorescent Dyes Freezing Glutamine GZMB protein, human Homo sapiens IL2RA protein, human Lymphocyte prisma Protoplasm Stem Cells Streptococcal Infections T-Lymphocyte T-Lymphocyte Subsets Transcriptional Activation Transcription Factor
2
Inhibition of VEGF-Receptor Binding
Not available on PMC !

Example 2

Evaluation of the Capability of Monoclonal Antibodies to Inhibit Binding of VEGF to its Receptor

An anti-VEGF antibody binds to VEGF to block the binding of VEGF to its receptors, VEGFR-1 and/or VEGFR-2, to be able to inhibit signal transduction through mediation of VEGF.

KLHa505 and KLHb1501 were separated and purified from the culture supernatants of the two positive clones using Protein G.

Next, IgG Fc-VEGFR-1 or IgG Fc-VEGFR2 was immobilized on a 96-well ELISA plate. After blocking with 2% bovine serum albumin, a purified antibody mixed with rhVEGF was added to the plate, followed by reaction at room temperature for 1 hour. A solution was prepared by mixing with rhVEGF, and then washed 3 times with 0.05% TWEEN® 20-containing TBS (TBS: 50 mM Tris-HCl (pH7.4), 500 mM NaCl; hereafter, referred to as “TBS-T”). Thereafter, through color development using rabbit anti-human VEGF polyclonal antibody-HRP, the rhVEGF content was determined.

As a result, it was demonstrated that the KLHa505 antibody competitively inhibits binding of VEGF to VEGFR-1 and VEGFR-2, and the KLHb1501 antibody competitively inhibits binding of VEGF to VEGFR-2 (FIG. 1).

That is, it was demonstrated in this Example that the antibodies of the present invention, KLHa505 and KLHb1501, can block VEGF-associated signal transduction.

US11872271B2. High-affinity anti-VEGF antibody KLHa505 (2024-01-16). ORDER-MADE MEDICAL RESEARCH INC. [JP], SANTEN PHARMACEUTICAL CO., LTD. [JP]. Inventors: Yasufumi Murakami [JP], Shigeki Mukoubata [JP], Hirotada Akiyama [JP], Koji Konomi [JP].
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Patent 2024
Antibodies Antibodies, Anti-Idiotypic Cardiac Arrest Clone Cells Enzyme-Linked Immunosorbent Assay FLT1 protein, human G-substrate Homo sapiens Immunoglobulins Monoclonal Antibodies Rabbits Serum Albumin, Bovine Signal Transduction Sodium Chloride Tromethamine Tween 20 Vascular Endothelial Growth Factor Receptor-2 Vascular Endothelial Growth Factors
3
Anti-CD22 Antibody Binding Profiles
Not available on PMC !

Example 3

FIG. 4 summarizes target binding activity of the anti-CD22 heavy chain-only antibodies described herein. Column 1 indicates the Clone ID number of the anti-CD22 heavy chain-only antibody. Column 2 indicates the binding affinity to protein (KD) measured in molarity. Column 3 indicates the dissociation constant of binding to protein (K-off rate) measured in seconds. Column 4 indicates binding to Daudi cells measured as fold over background MFI signal. Column 5 indicates binding to CHO cells stably expressing cyno CD22 measured as fold over background MFI signal. Column 6 indicates binding to CHO cells that do not express CD22 protein measured as fold over background MFI signal.

US11873336B2. Heavy chain antibodies binding to CD22 (2024-01-16). TeneoBio, Inc. [US]. Inventors: Shelley Force Aldred [US], Wim van Schooten [US], Heather Anne N. Ogana [US], Laura Marie Davison [US], Katherine Harris [US], Udaya Rangaswamy [US], Nathan D. Trinklein [US].
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Patent 2024
Binding Proteins Cell Lines Cells CHO Cells Clone Cells Immunoglobulin Heavy Chains Neoplasm Metastasis Proteins
4
Evaluating Anti-S100A8/A9 Antibodies for Lung Metastasis Suppression
Not available on PMC !

Example 4

Through use of a lung metastasis model of mouse breast cancer 4T1 cells, the lung metastasis-suppressing effects of anti-S100A8/A9 monoclonal antibodies were investigated.

In accordance with a protocol illustrated in FIG. 9, 1×105 mouse breast cancer 4T1 cells and 50 μg of each anti-S100A8/A9 monoclonal antibody (Clone Nos.: 45, 85, 235, 258, and 260) were simultaneously injected into the tail vein of five Balb/c nu/nu mice per group, and 2 weeks later, CT scans were performed. FIG. 10 shows the results for comparing typical CT images and the areas of tumor cells calculated from the CT images to those of a negative control group. As a result, it was recognized that Clone No. 45 showed a significant lung metastasis-suppressing effect.

US11858984B2. Anti-S100A8/A9 antibody and use thereof (2024-01-02). NATIONAL UNIVERSITY CORPORATION OKAYAMA UNIVERSITY [JP], NIIGATA UNIVERSITY [JP], NATIONAL UNIVERSITY CORPORATION GUNMA UNIVERSITY [JP], KAWASAKI GAKUENN EDUCATIONAL FOUNDATION [JP]. Inventors: Masakiyo Sakaguchi [JP], Shinichi Toyooka [JP], Shuta Tomida [JP], Kazuhiko Shien [JP], Hiroki Sato [JP], Rie Kinoshita [JP], Junichiro Futami [JP], Kota Araki [JP], Mikio Okazaki [JP], Eisaku Kondo [JP], Yusuke Inoue [JP], Akira Yamauchi [JP].
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Patent 2024
Breast Clone Cells Lung Lung Cancer Malignant Neoplasm of Breast Mice, Inbred BALB C Mice, Nude Monoclonal Antibodies Mus Neoplasm Metastasis Neoplasms Tail Veins X-Ray Computed Tomography
5
Mutagenesis and Lipid Production
Not available on PMC !

Example 6

Strain 5 was subjected to another round of mutagenesis with increasing concentrations and exposure time to 4-NQO (37 μM for 30 minutes at 28° C.). This population of cells was subsequently subdivided and grown in standard lipid production medium supplemented with a range of cerulenin concentrations (7-50 μM). Cells from all concentrations were pooled and fractionated over a 60% Percoll/0.15 M NaCl density gradient. Oil laden cells recovered from a density zone of 1.02 g/mL were plated and assessed for glucose consumption and fatty acid profile. One of these clones was subsequently stabilized and given the strain designation “Strain 6”.

US11873405B2. High oleic oil compositions and uses thereof (2024-01-16). Checkerspot, Inc. [US]. Inventors: Leon Parker [US], Kevin Ward [US], Scott Franklin [US], Mona Correa [US], Veronica Benites [US].
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Patent 2024
Cells Cerulenin Clone Cells Fatty Acids Glucose Lipids Microalgae Mutagenesis Oleic Acid Percoll Sodium Chloride Strains Triglycerides

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8
Lipofectamine 3000 by Thermo Fisher Scientific
Sourced in United States, China, Germany, Japan, United Kingdom, France, Canada, Italy, Australia, Switzerland, Denmark, Spain, Singapore, Belgium, Lithuania, Israel, Sweden, Austria, Moldova, Republic of, Greece, Azerbaijan, Finland
Lipofectamine 3000 is a transfection reagent used for the efficient delivery of nucleic acids, such as plasmid DNA, siRNA, and mRNA, into a variety of mammalian cell types. It facilitates the entry of these molecules into the cells, enabling their expression or silencing.
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Lsrfortessa by BD
Sourced in United States, United Kingdom, Germany, France, Australia, Canada, Belgium, China, Uruguay, Japan, Switzerland, Cameroon
The LSRFortessa is a flow cytometer designed for multiparameter analysis of cells and other particles. It features a compact design and offers a range of configurations to meet various research needs. The LSRFortessa provides high-resolution data acquisition and analysis capabilities.
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Pgem t easy vector by Promega
Sourced in United States, China, Germany, United Kingdom, Japan, France, Italy, Australia, Switzerland, Spain, Israel, Canada
The pGEM-T Easy Vector is a high-copy-number plasmid designed for cloning and sequencing of PCR products. It provides a simple, efficient method for the insertion and analysis of PCR amplified DNA fragments.

More about "Clone Cells"

Clone cells, also known as clonal cells, refer to a population of genetically identical cells derived from a single common ancestor cell.
These cells are widely utilized in biomedical research, including stem cell studies, cancer research, and genetic engineering.
Clone cells provide a valuable model system for investigating cellular processes, gene expression, and therapeutic development.
Researchers can leverage advanced AI-powered tools, such as PubCompare.ai, to optimize their clone cell experiments.
These tools enable researchers to easily locate and compare the best protocols from literature, preprints, and patents, enhancing reproducibility and streamlining the research process.
This is particularly useful when working with techniques like Lipofectamine 2000, a transfection reagent, or FBS (Fetal Bovine Serum), a common cell culture supplement.
The use of flow cytometry instruments, such as the FACSCanto II or FACSCalibur, is often integral to clone cell research, allowing for the analysis and sorting of cell populations.
Additionally, culturing clone cells often involves the use of media like DMEM (Dulbecco's Modified Eagle Medium) and the addition of antibiotics like Puromycin or Penicillin/Streptomycin to ensure cell health and purity.
For genetic manipulations, researchers may utilize vectors like the pGEM-T Easy vector, which can be used for cloning and propagating DNA sequences.
Furthermore, advanced transfection reagents, like Lipofectamine 3000, can be employed to efficiently introduce genetic material into clone cells.
The seamless integration of these tools and techniques, combined with the power of AI-driven protocol comparison, empowers researchers to drive their clone cell research forward, uncovering new insights and accelerating scientific discoveries.
By harnessing the power of clone cells and the latest advancements in research tools, scientists can make significant strides in fields ranging from regenerative medicine to cancer therapeutics.