...absence of unexpected DNA contaminants. The data is also used to determine recombination rates in FLEX/DIO/fDIO...Endotoxin contamination in vector preparations can alter the immunogenic properties of the final product...mycoplasma contamination. To date, Addgene has never had a case of mycoplasma contamination. In the event...is performed on viral genomes isolated from the final AAV preparation. Sequencing results are analyzed...particularly in large animal studies. Endotoxin contamination is minimized by using an endotoxin-free plasmid.... To minimize the immunogenic properties of the final vector preparation, the quantity of gram-negative...Takara, and is routinely tested for mycoplasma contamination using mycoplasma detection kits. The cell line...

...Verdoes Backbones for mouse recombinant antibodies A panel of recombinant monoclonal antibodies against... Collection Addgene distributes ready-to-use recombinant antibodies made from plasmids in our collection...alternative approach is to use synthetically produced recombinant antibodies, created by cloning antibody components...Some scientists choose to work with full length recombinant monoclonal antibodies (R-mAbs) expressed in mammalian...research applications from the NeuroMab/Trimmer Lab Recombinant mAb Collection . Once the plasmids are transiently...of production due to their small size. These recombinant antibody fragments can be used in experiments...tool kit for rapid cloning and expression of recombinant antibodies. Sci Rep. 2014 Jul 30;4:5885. Andrew...

...Antibodies produced this way are called recombinant antibodies. Recombinant antibodies are created by cloning... time and labor intensive. Recombinant expression validation A recombinant expression strategy is used...cultured immune cells, or through the use of recombinant plasmids. Animals - Antibodies produced in animals... of a single antibody with little variation. Recombinant plasmids - Antibodies can be produced from plasmids... and can either be used while suspended in the original media or isolated and placed in a buffer. Antibody... binding site. This process, known as V(D)J recombination, allows for a wide variety of antibodies to ...abundant isotype in blood, also found in tissues; predominant isotype in secondary immune response Monomer ...

...growth and proliferation. mTOR, a serine/threonine kinase, is found in two complexes, mTORC1 and mTORC2... and multiple familial cancers are caused by inactivating mutations in negative regulators of mTORC1. ...homolog LKB1 Also known as STK11; serine/threonine kinase 11 mTOR Mechanistic target of rapamycin NF1 Neurofibromin...S6K Also known as RPS6KB1; Ribosomal protein S6 kinase B1 TSC1/2 TSC1 TSC2 Tuberous sclerosis VHL Von ...LST8 homolog GSK3 GSK3A GSK3B Glycogen synthase kinase 3 MDM2 MDM2 proto-oncogene mSin1 Also known as ...as MAPKAP1; Mitogen-activated protein kinase associated protein 1 mTOR Mechanistic target of rapamycin ...; tumor protein p53 PDK1 Pyruvate dehydrogenase kinase, isozyme 1 PI3K Catalytic Subunits PIK3CA PIK3CB...

...to brainstorm practical strategies for using combinations of AAV-based molecular tools to achieve targeted...Cambridge, MA, 02139 Organizing Committee Chair: Dr. Melina Fan, Co-Founder and CSO, Addgene Organizing Committee...Dr. Constance Cepko, Harvard Medical School Dr. Melina Fan, Dr. Susanna Bachle, Dr. Karen Guerin, Dr. ...Research Campus Kenton Woodard Penn Vector Core Melina Fan, Karen Guerin, and Leila Haery from Addgene...Friday, September 21 9:00 AM Breakfast and Welcome Melina Fan 9:30 AM Cre and Flpo Katherine Matho 10:15 ...Regulation Ben Deverman 2:00 PM Cre-DOG and Combinatorial Strategies Connie Cepko 2:45 PM Break 3:00 PM...Field Kenton Woodard 4:00 PM Wrap Up & Next Steps Melina Fan/Leila Haery 5:00 PM Adjourn Meeting Preparation...

...backbone-based Gateway destination vectors (pGCS), bearing either amino- or carboxyl-terminal tags, including...zebrafish research. China Zebrafish Resource Center (CZRC) - A resource center based in China focused on collecting...Tol2 transposons, Cre-mediated site-specific recombination, morpholinos, zinc finger nucleases, TALENS,...vectors for cell-type specific conditional gene inactivation - Jeffrey Essner Lab GeneWeld Vectors for Targeted...origin in zebrafish can be found below: Optimized QF-binary expression system for use in zebrafish - Bret Pearson...zebrafish Whole organism lineage tracing by combinatorial and cumulative genome editing - Jay Shendure...toolkit - Tatjana Sauka-Spengler Lab. A genetic binary system for cell- and compartment-specific biotin...

...CDK6 Cyclin-dependent kinase 4 or 6 CHK1 Checkpoint kinase 1 CHK2 Checkpoint kinase 2 Cop-1 Ring finger ...factor 1 ATM ATM serine/threonine kinase ATR ATR serine/threonine kinase B99 G-2 and S-phase expressed 1...PMAIP1 p14ARF Cyclin-dependent kinase inhibitor 2A p21 Cyclin-dependent kinase inhibitor 1A p48 Damage-specific...to activate transcription, and they also have a dominant negative effect on functional p53 through oligomerization...

...Homologous Recombination (HR)? How long should the homology arms be for efficient recombination? The most...Dr. Le Cong, the first author of a seminal CRISPR article from Dr. Feng Zhang's Lab, has assembled a ...Plasmids Dr. Le Cong, the first author of one of the seminal articles from Dr. Feng Zhang's Lab ( Science , ...to learn more. CRISPR Design FAQs Homologous Recombination (HR) FAQs At the Bench FAQs CRISPR Design FAQs... to express the Cas9n (nickase). Homologous Recombination (HR) FAQs Can you summarize HR template vector...specific mutations or insertions by Homologous Recombination (HR), what is the length of each homology arm...destroy the BbsI sites, a double digest should discriminate between positive and negative clones. Clones...

...BbsI yes, cut, N-terminal S. pyogenes Zhang PX854 62886 Mammalian BbsI yes, cut, C-terminal S. pyogenes Zhang...BbsI yes, cut, N-terminal S. pyogenes Zhang PX852 62884 Mammalian BbsI yes, cut, C-terminal S. pyogenes Zhang...activate, N-terminal S. pyogenes Zhang PX856 62888 Mammalian BbsI yes, activate, C-terminal S. pyogenes...assembled into a single T-DNA binary vector of choice through Gateway recombination. Qi Multiplex Conditional...pyogenes GFP Vakoc U6>sgRNA(F+E) 59986 Other/Ciona intestinalis BsaI none S. pyogenes Christiaen p(U6a-BsaI-...Gersbach MuLE System Mammalian MultiSite Gateway recombination-based cloning of complex polycistronic lentiviruses...expression cassettes, which can be assembled into the binary CRISPR/Cas9 vectors in one round of cloning by ...

...Bacterial Expression mCardinal 604 659 17 5.3 27 min Prone to dimerization mCardinal-N1 - Mammalian Expression...Expression mCardinal-C1 - Mammalian Expression mCardinal-pBAD - Bacterial Expression mMaroon 610 652 - Monomer...eGFP pPD95_75 - C-terminal GFP for C. elegans expression pHT101-mCherry - N- or C-terminal mCherry for C....that are used extensively by life scientists. The original green fluorescent protein (GFP) was cloned in ...Davidson Lab Plasmids - Includes many N- and C-terminal fluorescent proteins Insect Sutherland Lab Plasmids...tagging with mCherry, mOrange, mCerulean pET GFP - C-terminal GFP for bacterial expression Davidson Lab Plasmids...Methods, September 2007, Vol. 4 No. 9, pp. 741-6 Shemiakina et al. : Nature Communications, November 2012...

...Find unique recombinant antibodies targeting families of cell surface and secreted proteins from a collaboration...Innovation (IPI) Collection! Here, you’ll find unique recombinant antibodies targeting families of cell surface...window) and an optimized sorting process. This combination allows IPI to generate antibodies to highly conserved...courtesy of IPI Browse Glypican Antibodies ID Recombinant Antibody Description Isotype Reactivity Recommended...courtesy of IPI Browse Integrin Antibodies ID Recombinant Antibody Description Isotype Reactivity Recommended...Browse Epitope Tag Antibodies IPI has developed recombinant versions of antibodies that recognize common ... tags and other common cellular proteins. ID Recombinant Antibody Description Isotype Reactivity Recommended...

...anti-kinesin recombinant scFvs . Niwa et al. Cytoskeleton. 2023 Apr 10. Explore the recombinant monoclonal... upper and lower motor neurons in the brain and spinal cord. Over time, a loss of motor neurons leads ...familial cases, usually inherited in an autosomal dominant manner, are associated with known genetic mutations...Huntington’s disease (HD) is a fatal autosomal dominant genetic disorder that causes the progressive degeneration... gene and late-onset AD. It is more likely a combination of gene variation and lifestyle or environmental...Saha et al. Nat Commun. 2023 Feb 2. Study the C terminal domain of TDP-43 to better understand its function... tools and emphasizes their rapid and broad dissemination to the research community. This collection highlights...

...methods and quality control processes for purified recombinant antibodies....webpage for further information. Production For recombinant antibodies (rAb) encoded by mammalian expression... buffer exchange using centrifugal columns. The final formulation buffer is phosphate-buffered saline ...sample is adjusted by diluting or concentrating to a final concentration of 0.9–1.1 mg/mL. Antibody concentration...software and a standard curve is generated. The final antibody concentration is calculated from the standard...performed in parallel with a previous lot of recombinant antibody or the hybridoma-derived equivalent ... Residual Plasmid Sanger Sequencing Purified recombinant antibody preps contain low levels of co-purified...

...) N-terminal Anti-MECP2 (M7443) mouse antibody from Sigma (Link opens in a new window) C-terminal Anti-MECP2...syndrome can be inherited. In females, random X inactivation results in approximately half of cells expressing...expressing mutant MECP2 . Skewing of these X inactivation ratios can affect disease severity. The MECP2...several functional domains, including: the N - t erminal D omain (NTD) the M ethyl B inding D omain (MBD...signal (not essential for localization) the C - t erminal D omain (CTD) The most common missense mutations...antibody from Sigma (Link opens in a new window) C-terminal Anti-MECP2 (D4F3) rabbit antibody from Cell Signalling...direction and degree of skewing of X-chromosome inactivation. J Med Genet . 44, 148–152. (Link opens in a...

...and shRNA against p53 in different gene/insert combinations A more efficient method to generate integration-free...NR5A2, and microRNA 302/367 in three different combinations of reprogramming factors Efficient germ-line...40. Hochedlinger MMLV-derived Retrovirus Mouse Original set of Yamanaka factors for generating mouse iPS... and c-Myc from polycistronic cassettes KLF4 N-terminal variance modulates induced reprogramming to pluripotency...beta-cells. Nature. 2008 Aug 27. Melton Pancreatic Acinar Cells induced Beta Cells Adenoviral Mouse Long-...pancreatic beta cells generated by lineage conversion of acinar cells. Nature Biotechnology. 2014 Dec;32(12):1223... of mouse and human fibroblasts into functional spinal motor neurons. Cell Stem Cell. 2011 Sep 2;9(3):...

...or C-terminal tagging in Drosophila cells. N terminal tagging in Drosophila cells 3.2 MB C terminal tagging...deposited a plasmid which introduces an N- or C- terminal affinity tag (3xFLAG-2xSTREP) on endogenous genes...untagged SpCas9 (Addgene 41815 or #44719 ) in combination with gRNA_AAVS1-T2 (Addgene #41818) or using ...modification. This system can be used to create C- and N-terminal epitope tags. The plasmids in the following articles...selection markers are available. An improved FLP recombinase expression vector allows for efficient marker...

...developing mouse retina. To aide in their studies of the regulatory networks involved in retinal development...plasmids to study the regulatory networks involved in retinal development. Depositor...990 KB) Plasmid and Sequence Derivative Sources Original pX458 backbone ( Addgene #48138 ) dCas9 + KRAB-MeCP2...Initiation of Otx2 expression in the developing mouse retina requires a unique enhancer and either Ascl1 or ...Simultaneous deletion of Prdm1 and Vsx2 enhancers in the retina alters photoreceptor and bipolar cell fate specification...

...Expression of the tet-off transactivator (tTA) 2 Gradinaru 99120 pAAV-ihSyn1-tTA Inducible Synapsin promoter...feedback loop for amplified tTA expression. 1 Gradinaru 99121 pAAV-ihSyn1-DIO-tTA Cre-dependent and inducible...feedback loop for amplified tTA expression. 1 Gradinaru 117383 TRE-DIO-eYFP Cre-dependent and Tetracycline-inducible...Cre-dependent, Tet-inducible expression of EYFP 1 Gradinaru Tools for Affinity Purification These AAV encode...tdTomato and synaptophysin-EGFP for labeling of axon terminals. 1 Zeng 71760 pAAV hSyn FLEx mGFP-2A-Synaptophysin-mRuby... and synaptophysin-mRuby for labeling of axon terminals. 1 Luo 60658 pAAV-EF1α-F-FLEX-mNaChBac-T2A-tdTomato...

...nickase or catalytically inactive “dead” Cas9 (dCas9) to a cytidine deaminase like APOBEC. Base editors...creating an A to G (or T to C) change. Adenine DNA deaminases do not exist in nature, but have been created... of the Escherichia coli TadA, a tRNA adenine deaminase. Like cytosine base editors, the evolved TadA ...modifying the linker region between Cas variant and deaminase to adjust the editing window, or adding fusions... Cas9 is fused to activation-induced cytidine deaminase (AID). Want more information on the wide variety...

... and H 2 O to final volume of 50 μl. This reaction may be scaled down to a smaller final volume if necessary...video, you can find the protocol section from the original publication below. You can also download the full-text...., sgRNA-B in Table 4 ) and do not add C to the final position of the reverse complement oligo. In this...Buffer, 6.5 μl ddH 2 O, and 0.5 μl T4 Polynucleotide Kinase (PNK) (10,000 U/ml). NOTE: Phosphorylated oligos...contains 50 μl cell culture media per well for a final volume of 150 μl. This facilitates screening by ...research. Gehl J. Acta Physiologica Scandinavica . 2003. PubMed . Purifying plasmid DNA from ...