...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 ...

...3rd 6-25 11,108 Borodina Yeast Transporter Libraries 153101-153106 Knockout Yeast Borodina N/A 1 Varies ...Human Ebert 3rd 4 2,852 Bonifacino Lab Human ubiquitination-related proteins CRISPR KO library 174592 Knockout...contained in the library represent all 4096 (4 6 ) combinations of 6 nucleotides. Chong Zhang E. coli Genome-wide... Human Weissman 3rd 6 Varies Enriched subpools (kinase, nuclear, ribosomal, cell cycle) 51043 — 51048 ... 84,963 Human CRISPR Knockout Pooled Library (Gattinara) 136986 Knockout Human Doench 3rd 2 40,964 Ingolia...Library 140633 Knockout Human Wei 3rd 3 58,630 Human Kinase Domain-Focused CRISPR Knockout Library 117725 Knockout...

...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...

...vector for insertion of N-terminal domain (SgrAI or AgeI with ClaI) and C-terminal domain (FseI PacI). Grow...modules are orthogonal, researchers can use any combinations of modules as is, allowing plug-and-play application...

...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...

...between mouse and human Jonathan Ting Viviana Gradinaru 163509-PHPeB CN1839-rAAV-hSyn1-SYFP2-10aa-H2B-...scRNA-seq applications. Jonathan Ting Viviana Gradinaru 164450-PHPeB CN1851-rAAV-hI56i-minBglobin-iCre...forebrain GABAergic interneurons. For use in combination with Cre-dependent transgenic mouse lines or ...dependent viral vectors. Jonathan Ting Viviana Gradinaru 37825-CAP-B10 pAAV-CAG-GFP AAV expression of GFP...GFP from the CAG promoter Edward Boyden Viviana Gradinaru 37825-CAP-B22 pAAV-CAG-GFP AAV expression of GFP...GFP from the CAG promoter Edward Boyden Viviana Gradinaru 37825-CAP-MaCPNS1 pAAV-CAG-GFP AAV expression ...GFP from the CAG promoter Edward Boyden Viviana Gradinaru 37825-CAP-MaCPNS2 pAAV-CAG-GFP AAV expression ...

...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...

...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...

...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 ...

...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...

...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...

...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):...

...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...

... 5, rg*, PHP.eB Gradinaru 104061 CAG-NLS-GFP CAG NLS-GFP Constitutive PHPeB Gradinaru 105530 pAAV.CMV....NLS-mRuby2 Constitutive 1, 2, 8, 9, rg*, PHP.eB Gradinaru 100896 pAAV.GFA104.PI.eGFP.WPRE.bGH GFA104 EGFP...eYFP hSyn eYFP Constitutive 5, 8, 9, rg*, PHP.eB Gradinaru 131000 pAAV-CaMKIIa-mScarlet CamKIIa mScarlet ...pAAV-CAG-DIO-EYFP CAG EYFP Cre dependent PHP.V1 Gradinaru 83895 pAAV-hDlx-Flex-GFP-Fishell_6 Dlx GFP Cre...pAAV-CAG-fDIO-mNeonGreen CAG mNeonGreen Flp dependent 1 Gradinaru INTRSECT Constructs 55650 pAAV-hSyn Con/Fon EYFP...

... 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 ...

...cloned, expressed and validated in recombinant form. The recombinant monoclonal antibodies or R-mAbs have...149451 Anti-GluK5/Grik5/KA2 kainate receptor [N279B/27R] GluK5/Grik5/KA2 kainate receptor Rat Mouse IgG2a...Anti-MMP9 matrix metalloproteinase-9 precursor [L51/82R] MMP9 matrix metalloproteinase-9 precursor Rat ..., followed by the SV40 polyA tail (SV40 pA). Recombinant Monoclonal Antibody Plasmid Collection The following... Mouse IgG1 Addgene distributes ready-to-use recombinant antibodies made from plasmids in our collection...channel scFv [D4/154] D4/154 scFv Kv2.1 K+ channel recombinant Mouse 206788 Arx scFv [N411/51] N411/51 scFv ...al. 2019. A toolbox of IgG subclass-switched recombinant monoclonal antibodies for enhanced multiplex ...

...Human Kinase ORFs Genome Engineering Guide Empty-Backbone Resource Mitogen-activated protein kinases (MAPKs...extracellular signal-regulated kinases (ERKs), are serine/threonine-specific protein kinases. They regulate diverse...Browse mitogen-activated protein kinase (MAPK) plasmids. Plasmid...