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...concentration!). Sample Data Figure 1: The plate was coated with serial dilutions of human recombinant purified...D7]. Plates were incubated with HRP-linked anti-Human IgG secondary antibody for 2 hours, reacted with... Protocols Antibody Validation Using the Indirect ELISA Method Antibody Validation Using the Indirect ...ImmunoSorbent Assay (ELISA) against a purified antigen to validate an antibody. This protocol will describe how to...that the protocol may need to be adjusted to accommodate slight differences between products. Addgene ...

...backbone used by the RNAi consortium for targeting human and mouse genes....built a library of shRNAs directed against 15,000 human and 15,000 mouse genes. Addgene is working with ...www.addgene.org/10878/ . Description Vector Element U6 Human U6 promoter drives RNA Polymerase III transcription...preintegration complex in the transduced cells. hPGK Human phosphoglycerate kinase promoter drives expression...209-216. PubMed . A lentiviral RNAi library for human and mouse genes applied to an arrayed viral high-content...Institute's TRC shRNA Design Process . To minimize degradation of off-target mRNAs, use NCBI’s BLAST program...days. h. Assay infected cells. The following recommendations are guidelines for the number of days you should...

...Consortium Healthy human cells Imaging (IHC, ICC, IF, etc.)  Data repository From the Human BioMolecular...Any Data repository   YCharOS Human proteins Immunoblot (western blot), IP, and IF. Data repository...others) Data repository From the Human Cell Differentiation Molecules group Addgene Data Hub Any...imaging Data repository See blog post mAb3D Atlas Mouse and human brain  IHC Data repository... Any Data repository From the Frederick National Library for Cancer Research Human Luekocyte... include validation data CiteAb Any Any Search engine May include validation data  Biocompare...websites that can help: Antibody data repositories, which share validation data, and antibody search engines...

...with microbes that don’t cause disease in healthy humans, for example, non-pathogenic E . coli . BSL-2 is...with chemicals, first review their material safety data sheets (MSDS). While You Work Dispose of sharps ...

...backbone used by the RNAi consortium for targeting human and mouse genes. Virus Protocols for titering and...antibodies to detect antigens in cells Antibody Validation Using the Indirect ELISA Method Run an indirect...

...N222/6R] Clcn4 Human Mouse IgG2a 114475 Anti-PARIS/ZNF746 [N196/16R] PARIS/ZNF746 Human Mouse IgG2a 114476...Mucolipin-3 Human Mouse IgG2a 114478 Anti-GABA(A)R, Epsilon [N413/67R] GABA(A)R, Epsilon Human Mouse IgG2a...Anti-Stonin-2 [N346/9R] Stonin-2 Human Mouse IgG2a 114485 Anti-NSD3 [N348/82R] NSD3 Human Mouse IgG2a 114486 Anti-Flrt3...Menke's disease protein) Human Mouse IgG2a 114503 Anti-LRRK1 [N341/37R] LRRK1 Human Mouse IgG2a 114504 Anti-TrpC4...Anti-Malin [N85/18.1R] Malin Human Mouse IgG2a 114514 Anti-DNMT3L [N117/9.5R] DNMT3L Human Mouse IgG2a 114515 Anti-GABA...N147/6.1R] Pan-QKI Human Mouse IgG2a 114525 Anti-FGF13/FHF2 [N91/27.1R] FGF13/FHF2 Human Mouse IgG2a 114527...1.1R] Neurexin-1-Beta Human Mouse IgG2a 114533 Anti-Laforin [N84/37.1R] Laforin Human Mouse IgG2a 114536 ...

...conditions commonly encountered in human gene therapy trials. Human Gene Therapy Methods, 26(2), 71–76...Researcher Perspectives on Data Sharing in Deep Brain Stimulation. Frontiers in Human Neuroscience, 14(December...Vector-Mediated Gene Delivery to the Hypothalamus. Human Gene Therapy, 21(6), 673–682. https://doi.org/10.1089...Adeno-Associated Virus Serotype 9 (AAV9) Preparations. Human Gene Therapy Methods, 30(6), 214–225. https://doi.org... AAV Data with Addgene's New AAV Data HubCelebrate Open Data Day with Addgene’s AAV Data Hub!Data Freedom...asking scientists what data would improve our AAV Data Hub.   The Addgene Data Hub is here to help!  ...through DataCite before being made publicly available through the Data Hub. These minimum data standards...

... pFETCh_Donor CEBPB Human FLAG pFETCh_CEBPB PX458_CEBPB_1 PX458_CEBPB_2 AHR Human FLAG pFETCh_AHR PX458...AHR_1 PX458_AHR_2 ARID4B Human FLAG pFETCh_ARID4B PX458_ARID4B_1 BCL6_iso1 Human FLAG pFETCh_BCL6_iso1 PX458..._BCL6_iso1_2 CEBPG Human FLAG pFETCh_CEBPG PX458_CEBPG_1 PX458_CEBPG_2 RAD21 Human FLAG pFETCh_RAD21 PX458...PX458_RAD21 GABPA Human FLAG pFETCh_GABPA PX458_GABPA_1 PX458_GABPA_2 GABPB1_iso2 Human FLAG pFETCh_GABPB1...iso2 PX458_GABPB1_iso2_1 ATF1 Human FLAG pFETCh_ATF1 PX458_ATF1 CEBPA Human FLAG pFETCh_CEBPA PX458_CEBPA...PX458_CEBPA_2 CREB1 Human FLAG pFETCh_CREB1 PX458_CREB_1 PX458_CREB_2 TGIF2 Human FLAG pFETCh_TGIF2 PX458...PX458_TGIF2_2 SSRP1 Human FLAG pFETCh_SSRP1 PX458_SSRP1_1 PX458_SSRP1_2 ZNF146 Human FLAG pFETCh_ZNF146...

...Description Human SLC Activation Library 132561 Superti-Furga CRISPR activation library targeting human SLC proteins...proteins. Human SLC Knockout Library 132552 Superti-Furga CRISPR knockout library targeting human SLC proteins...Inducible barcoded overexpression library of human SLC proteins. Human Transporter Knockout Libraries 213695 ...) are the largest family of transporters in the human genome with approximately 450 identified members...collection contains codon-optimized ORF sequences for human SLCs in Gateway-compatible pENTR vectors (pDONR221... knockout library targeting mouse SLC proteins. Human SLC Overexpression Library 213694 Superti-Furga ...Superti-Furga CRISPR knockout library targeting human transmembrane transporters. Feedback Please send...

...the Human Leucocyte Differentiation Antigens (HCDM) workshop approach, which focuses on validating antibodies...antibody validation efforts and information provided by external organizations like the Human Cell Differentiation...Bradding, P. (2019). Validation of antibodies for the specific detection of human TRPA1. Scientific Reports...antibodies against markers of the human blood leukocyte populations, as well as a pragmatic approach to...characterizing cell surface molecules expressed on human blood leukocytes, including antibodies against these...Roach, K. M. (2021). TGFβ1 induces resistance of human lung myofibroblasts to cell death via down-regulation...target type. To start the validation process, one must first confirm that a candidate antibody can detect the...

...translate AID to yeast, as well as mammalian and human cell lines (Nishimura et al., 2009, Natsume et al... your protein of interest to create conditional human cell lines. Lastly, the Kanemaki lab demonstrated...conversion in the target sites adjacent to an NG PAM in human cells. The increased versatility of the engineered... for degradation using AID Article contributed by Alyssa Cecchetelli Auxin Inducible Degradation (AID)...publicly available metagenomic data including the Tara Oceans Project datasets. This study demonstrates the...available metagenomic datasets with biophysical studies and the potential of mining datasets for research and...interest fused to a degron, a domain that induces degradation. In plants, auxin binds to TIR1 of the SCF-TIR1...

... Tagging in Human Cells on the Addgene Blog. Plasmids for Endogenous Gene Tagging in Human Cells Find ...passaging of human iPS cells (Link opens in a new window) RNP transfection for gene editing human iPS cells...fluorescently tagged proteins of key cellular structures in human cell lines. Depositor... opens in a new window) of fluorescently-tagged human induced pluripotent stem cell (hiPSC) lines. The...fluorescently tagged proteins of key cellular structures in human cell lines of their choice. For a full description...). Systematic gene tagging using CRISPR/Cas9 in human stem cells to illuminate cell organization. Mol ...Institute for Cell Science data portal (Link opens in a new window) , including: data and images from the hiPS...

...elegans ), fruit fly, zebrafish, mouse, rat, and human. Developed by the Stanley Qi Lab . CCTop - CRISPR...location, and genome annotation. Available genomes are human (hg19 & hg38), mouse (mm10), and yeast (strain w303... a set of similar sequences. Organisms include human, mouse, rat, chicken, frog, zebrafish, fly, worm...on-target activity for selected target(s). Uses human, mouse, and rat reference genomes. Hosted by the Broad...meningitidis Cas9 PAMs. Currently supports: mouse and human. Developed by the George Church Lab . CRISPR Optimal... GFP (pSIR-GFP) , DsRed (pSIR-DsRed-Express2) , human CD2 (pSIR-hCD2) PDF, 108 KB Fujii iChIP/enChIP to...GABPA, and RAD21 PDF, 138 KB Musunuru CRISPRs in human pluripotent stem cells pCas9_GFP ; gRNA empty vector...

...previously observed instance of human serine phosphorylation ( Rinehart Human Serine Phosphopeptide Library...adapted from the synthetic human phosphoproteome. It subdivides the 110,139 human phosphoserine site library...phosphosite library can then be used to identify candidate human substrates of kinases of interest by mass spectrometry...pSerOTS-C1* (V70) (DOCX, 551 KB) . The synthetic human phosphoproteome The Rinehart lab synthesized 110,139.... A multi-purpose, regenerable, proteome-scale, human phosphoserine resource for phosphoproteomics . Nature...iSPI, rEcoli XPS, pSerOTS-C1* (V70) (DOCX, 551 KB) Human Serine Phosphopeptide Library described in: Barber... Isaacs, F. J., & Rinehart, J. (2018). Encoding human serine phosphopeptides in bacteria for proteome-...

... 2013) Express human DNMT1 with a Myc tag. (Li et al. J Biol Chem. 2006) Express human ATM with Flag and.... (Marquart et al. Nat Methods. 2024) Base edit human mitochondrial DNA using mitoBEs . (Yi et al. Nat...Cell. 2021) Conduct multimodal genetic screens in human iPSC-derived neurons with a CRISPRi-based platform...and efficient generation of oligodendrocytes from human induced pluripotent stem cells. (Ehrlich et al. ... . Find plasmids for endogenous gene tagging in human cells from the Allen Institute for Cell Science ...new window) Michael J Fox Foundation (Link opens in a new window) A foundation dedicated to finding a cure...Plasmids provided by the foundation can be found on our Michael J. Fox Foundation Plasmid Resource page ...

... ) Rats ( Tesson et al., Nat Biotechnol. 2011 ) Human somatic cells ( Miller et al., Nat Biotechnol. 2011... ; Reyon and Tsai et al., Nat Biotechnol 2012 ) Human pluripotent stem cells ( Hockemeyer et al., Nat ...Biotechnol. 2011 ) TALE transcriptional activators in human cells ( Maeder et al., Nat Methods 2013 ) More detailed...Endogenous Zebrafish Genes TALENs for Endogenous Human Genes TALENs for the EGFP Reporter Gene Additional...REAL-Fast and FLASH, including detailed and up-to-date protocols, can be found on the TALengineering website...

...Developing Human Brain - includes developmental transcriptome information, prenatal LMD microarray data, ISH...ISH image data, and anatomic reference atlases of prenatal and adult human brain (Link opens in a new window...Collection and Resource Center MECP2 Animal Models Human Cell Lines CRISPR Tools Plasmids Resources The (..., suggesting that restoring MECP2 expression in humans could be curative. (Link opens in a new window)...common mutations that recapitulate Rett syndrome in humans. MECP2 knockout rats are available from (Link opens...conditional deletion of Xist Unpublished Joost Gribnau Human Cell Line Models Induced pluripotent stem cells ...mammalian or bacterial) Insert species (mouse or human) Mutation ID Plasmid Description Gene/Insert Mutations...

...confident in your data. This is a good option for applications like immunohistochemistry on human tissues, when...favorite protein in human tissues. You know that there is a wealth of transcriptomic data from this sample...your gene of interest across a set of human tissues.  From this data, you see that there is high RNA expression... major takeaway from these recommendations is: validate, validate, validate. This doesn’t necessarily ...extensive validation experiments for every antibody that you receive; citing existing validation data can be...extra validation experiments yourself.  Goal of Antibody Validation  As you assess existing data or plan...appropriately validated so that you can have confidence in your results. But what is appropriately validated? This...

...Assembly by ligation-independent cloning (LIC). Validated in human cells. Veit Hornung Musunuru/Cowan Lab TALEN...Toolbox PCR/Golden Gate cloning method. Optimized for human expression. Feng Zhang LIC TAL Effector Assembly... for deletion mutagenesis and gene targeting in human and mouse pluripotent stem cells. 48706 pTAL7b Joung... TALE repeats (C, T, and G) and the full length human LSD1 at the carboxy-terminus of the protein. 49043...in labeling specific repetitive DNA sequences in human chromosomes Do you have suggestions for other plasmids...Golden Gate TALEN 2.0 Golden Gate cloning method. Validated in multiple organisms. Dan Voytas Adam Bogdanove...Engineering Reagents Assembly via serial ligation. Validated in zebrafish somatic cells. Keith Joung Zhang ...

...below. Pooled genome-wide human SAM library SAM can be combined with a human genome-wide library to activate...These plasmids contain two expression cassettes, a human codon-optimized SpCas9 or SpCas9n, and the single...This plasmid contains a single cassette expressing human codon-optimized SpCas9; sgRNA can be co-transfected...(Ran et al., Nature Protocols 2013). Individual human codon-optimized SpCas9 plasmids can be ordered via...GeCKO are pooled lentiviral CRISPR libraries (both human and mouse) for performing genome-wide knockout screens...smaller Cas9 from S. aureus (SaCas9) that has been human codon-optimized, and packages both the SaCas9 gene...These plasmids contain two expression cassettes, a human codon-optimized SaCas9 driven by either the ubiquitous...