We narrowed to 264 results for: his-tag
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TypeProtocol...add 57 mL of 1.46 M sorbitol, 7.5 mL HI-FBS, and 7.5 mL 100X glutaGRO, 7.5 mL of 7.5% Sodium Bicarbonate... 2, Corning 3269, 1272 cm 2 Heat-inactivated FBS (HI-FBS) Pro-Tip Different brands and lots of FBS can... sorbitol (optional) : DMEM low glucose + 1% v/v HI-FBS + 0.1 M sorbitol In our hands, we have found ...sterile 250 mL bottle. Aliquot 774 mL of DMEM + 2% HI-FBS into a sterile 1 L bottle. Add each plasmid DNA...solution to the bottle containing 774 mL of DMEM + 2% HI-FBS. Mix well. Take the CS5 out of the incubator...L-alanyl-L-glutamine (or alternative stable glutamine such as glutaGRO, Corning 25-015-CI) Opti-MEM, Thermo Fisher 31985...L-alanyl-L-glutamine (or stable alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add ...
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Bacterial Expression Systems
TypeCollection...N-terminal 6x His tag and an MBP tag. Compatible with Gateway cloning. pDEST-periHisMBP 11086 Tac (lactose...pET-HIS-Sangamo 40786 TALEN Gang Bao Gateway cloning and expression of an N-terminally His-tagged TALEN...-Sangamo-His 40787 TALEN Gang Bao Gateway cloning and expression of a C-terminally His-tagged TALEN fused...artifacts!), photoactivatable tags for super resolution imaging, epitope tags like His for easy western blotting...recombinant protein with an N-terminal 6x His tag and an MBP tag. Contains a signal sequence to localize...different combinations of His and Strep tags as well as N-terminal PelB tags for localization to the periplasm...and C-terminal Strep, His, MBP, TRX, GST, SUMO, MISTIC, and LSL tags as well as tags for protein localization... -
Empty Backbones - Choosing Your Perfect Plasmid Backbone
TypeCollection...with N-terminal Myc tag and attB for Drosophila transgenesis His Epitope tag pEZYmyc-His - C-terminal ...fusion protein. Read more about epitope tags and protein tags . Tag or Fusion Protein Common uses...bacterial expression; His,GST tag pE2c - Modified pENTR vector for C-terminal 3x HA tag fusion with your gene...bacteria with N-terminal His-TrxA-TEV tag NusA Protein purification pNIC-HIS6-NUSA-TEV-GG - Cloning of...collection page Return to top Epitope Tag or Fusion Protein Tags and fusion proteins are excellent tools...stop codon for C-terminal tags and omit the start codon for N-terminal tags. And when you are designing... 2xFLAG tag in pCS for a variety of systems, including Xenopus HA Epitope tag pcDNA3.1... -
Fluorescent Protein Guide: FRET
TypeCollection...optimized CyPet pCyPet-His Cyan Bacterial Expresses CyPet with C-terminal His tag SCFP3A Cyan Mammalian...optimized YPet pYPet-His Yellow Bacterial Expresses YPet with C-terminal His tag SYFP2 Yellow Mammalian...Bacterial Expresses Aquamarine with N-terminal His tag pAquaN1 Cyan Mammalian Expresses mammalian optimized... suitable for creating individual fluorescently tagged proteins to study protein-protein interactions ... -
To Each HIS Own
TypeBlog Post...purification based on the polyhistidine tag. What is the polyhistidine tag? The polyhistidine tag is a string of ...Benefits of HIS tags In addition to providing a way to purify a protein of interest, polyhisitidine tags can ...low level, try a longer histidine tag to increase capture. Longer histidine tags are also recommended if... isolated. Prior to the polyhistidine tag, alternative fusion tags were used, such as S. aureus’... conditions. Find polyhistidine tag plasmids here! Where should I place the tag and how many do I need...plasmid containing a polyhistidine tag, you’ll need to decide whether to add the tag to the N- or C-terminus... however, tag placement can affect protein function. B, The string of histidines in the tag is typically... -
CRISPR History and Development for Genome Engineering
TypeCollection... (CAPTURE.) Tag : Multiple methods make it easier to tag endogenous loci with epitope tags or fluorescent...include mutations seen in human patients, protein tags, or loxP/FRT sites, among others. Homology-directed... not occur in mammalian cells. Purify : Epitope-tagged dCas9 can also be used to purify a genomic locus... EM, Myers RM. 2015. CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins. Genome Res . ... CRISPR CRISPR History CRISPR History and Development for Genome Engineering ... -
Neurodegeneration Plasmid Collection
TypeCollection...Trono 14666 pQE9-His-p97(wt) VCP His T5 ALS Graham Warren 14667 pQE9-His-p97(QQ) VCP His T5 ALS Graham Warren...17229 pQE9-His-p97deltaD2 VCP His T5 ALS Hemmo Meyer 17230 pQE9-His-p97deltaD2(K251A) VCP His T5 ALS Hemmo...0005 VCP His T7 ALS Axel Brunger 12339 pBD-0071 VCP His T7 ALS Axel Brunger 12344 pBD-0010 VCP His T7 ALS...0016 VCP His T7 ALS Axel Brunger 12346 pBD-0013 VCP His T7 ALS Axel Brunger 12347 pBD-0006 VCP His T7 ALS...0009 VCP His T7 ALS Axel Brunger 12349 pBD-0070 VCP His T7 ALS Axel Brunger 12350 pBD-0069 VCP His T7 ALS...0008 VCP His T7 ALS Axel Brunger 12352 pBD-0007 VCP His T7 ALS Axel Brunger 12353 pBD-0018 VCP His T7 ALS...0020 VCP His T7 ALS Axel Brunger 12355 pBD-0022 VCP His T7 ALS Axel Brunger 12356 pBD-0024 VCP His T7 ALS... -
Malate Dehydrogenase CUREs Community Collection
TypeCollection... cloned into an IPTG-inducible His-tag expression vector. All His-tags are cloned at the C-terminus of...inexpensive to assay, and easy to purify using histidine-tagged constructs and routine protein and molecular...map, a TEV cleavage site is present between the 6xHis-tag and the coding region of MDH. For detailed clone... -
Antibodies 101: Affinity Tags
TypeBlog Post...covered! His His tags, often called polyhistidine tags, typically include six consecutive His residues...Diagram of His-tagged protein purification work flow by nickel column. Affinity tags vs. other ...other tags There are many different types of tags, including epitope tags (FLAG, Myc, etc.) and fluorescent...blog Plasmids 101: Protein tags Antibodies 101: Epitope tags To Each HIS Own ...planning, affinity tags can make protein purification a cinch. Types of affinity tags, the proteins they...are all reviewed here. What are affinity tags? Affinity tags are used primarily as protein purification... common tags available. Maltose binding protein Maltose binding protein (MBP) is a 45 kDa tag which can... -
Fluorescent Protein Guide: Empty Backbones
TypeCollection...Includes tagging with CFP, YFP, and mRFP Tsoulfas Lab Lentiviral Plasmids - Fluorescently tag your gene...empty plasmid backbones with different fluorescent tags for you to create fusion proteins with your gene...Expression pCyPet-C1 - Mammalian Expression pCyPet-His - Bacterial Expression CYPet-N1 - Mammalian Expression...dimerization pCEP4YPet-MAMM - Mammalian Expression pYPet-His - Bacterial Expression YPet-N1 - Mammalian Expression...Monomer pmiRFP670-N1 - Mammalian Expression pBAD/His-miRFP670 - Bacterial Expression iRFP670 643 670 13...Monomer pmiRFP703-N1 - Mammalian Expression pBAD/His-miRFP703 - Bacterial Expression iRFP702 673 702 8... -
Plasmids 101: Protein tags
TypeBlog Post...affinity tag is the polyhistidine (His) tag. Small and unlikely to affect function, His-tagged proteins...usually using a Ni2+ column. Like other affinity tags, a His tag can be fused to either the N- or C-terminus...Combo and cleavage tags Frequently, a single tag is not enough. What if you need one tag to increase solubility... to a purification scheme, His tags are frequently combined with other tags for dual-labeling. The problem...to all tags, but rather a quick overview of why scientists use tags, with a few time-tested tags and fusion...length a polyhistidine tract does not greatly alter the size of the tag. Table 1: Common protein tags ...keep your protein soluble. Tags for affinity and purification An affinity tag, generally a relatively small... -
The Michael Davidson Collection: One-Stop Shop for Fluorescent Proteins
TypeBlog Post...Tallahassee Democrat in a story about his life’s work and the stage IV lung cancer that has now prompted...State University has been the go-to place. In 2012, his National High Magnetic Field Lab worked with an impressive... all those collaborations over the years, he and his colleagues built a Molecular Expressions collection...spectra. A comprehensive list of cell markers all tagged with mEmerald. Plasmids to monitor protein-protein...available targets since the Davidson lab always tagged new proteins with mEmerald first, Baird says. From...cell membrane, this collection has fluorescently tagged versions of many proteins known to localize to ...prompted him to deposit the tools he and his team created with the research community at large, "[Fluorescent... -
Institute for Protein Innovation
TypeCollection...Protein C, DYKDDDDK, Rho, His, Biotin, EE, GCN4 and Strep. Please visit our Tags and Other Markers Antibody... Glypican Antibodies Integrin Antibodies Epitope Tag Antibodies Antibody Plasmids Welcome to the Institute...proteins (glypicans and integrins) and common epitope tags, as well as antibody plasmids to create or express...Reactivity Recommended Applications Browse Epitope Tag Antibodies IPI has developed recombinant versions...versions of antibodies that recognize common epitope tags to help purify, detect, or localize particular proteins...s full catalog of antibodies that target epitope tags and other common cellular proteins. ID Recombinant... -
Special Delivery: Fluorophore Targeting for FRET Studies
TypeBlog Post...which can be carried out using Addgene plasmids. His-tag labeling reagents You may already have a FRET acceptor-binding...for protein purification: the NTA binds to the His tag on your protein and the coupled Cy fluorophore ...C-terminal fusion of a tetracysteine tag. In addition, since these tags are so small, one can insert them... between 80% and 10%. C Cy3NTA and Cy5NTA bind His tags similarly but with differing R0 values, thereby...interest: you just don’t know it yet! The poly-histidine tag often used for protein purification is a reliable..., as well as by extending the length of the histidine tag from 6 to 10 residues. While these Cy/NTA conjugates...incorporation of fluorescent probes into protein: hexahistidine-tag-mediated fluorescent labeling with (Ni2+: ... -
Course-Based Undergraduate Research Experiences (CUREs)
TypeBlog Post...same kinds of opportunities for his students. But he also found that his teaching responsibilities left...limited time for one-on-one mentoring. His solution? Bringing his research into the classroom. I had the...to hear about how he developed his program. Initially, Provost made his labs completely open-ended, allowing...bacteria, purified them to near homogeneity using a 6XHis tag, and compared their specific activities by measuring...purification and characterization of a fusion protein: Histidine tag, malate dehydrogenase, and green fluorescent...San Diego), decided to take a different approach. His own most impactful learning experiences happened ... opportunity to talk with Provost recently about his development of what is now commonly termed a Course-based... -
BeHeard Award 2018: Diseases of Glycosylation, Arginine Mutagenesis, & Neural Development
TypeBlog Post...Campus in Aurora, Colorado, USA. The primary focus of his lab’s research is the mechanisms by which proteins...Gronostajski is a professor at the University of Buffalo. His research focuses on understanding the structural ...research on diseases of glycosylation, arginine mutagenesis, and neural development. Paula Videira: Congenital...as reporters of NFIX function. They will then mutagenize the NFIX gene and test whether variants of unknown... -
Tips for Using FRET in Your Experiments
TypeBlog Post...optimized CyPet pCyPet-His Cyan Bacterial Expresses CyPet with C-terminal His tag SCFP3A Cyan Mammalian...optimized YPet pYPet-His Yellow Bacterial Expresses YPet with C-terminal His tag SYFP2 Yellow Mammalian...Bacterial Expresses Aquamarine with N-terminal His tag pAquaN1 Cyan Mammalian Expresses mammalian optimized...interactions in living cells using a microscope. Historically, molecular interactions were detected by indirect... -
Quick, Versatile Plant Transgenesis with GreenGate Plasmids
TypeBlog Post...promoters, N-terminal tags, coding sequences of the gene of interest, C-terminal tags, plant terminators ...science lab.” Introducing GreenGate As Lohmann and his colleagues describe in the PLoS ONE paper that introduced... it no longer works, just trash it.” Lohmann and his colleagues including Joachim Forner now expect and...aptly named GreenGate – offers all of those same advantages to researchers working in plant model systems... -
#DeckTheLab to Win a Tasty Winter Treat This Holiday Season
TypeBlog Post..., too! Bluegene is eager to decorate the lab but his short arms are causing him some difficulty… To give...celebrating the holidays and tweeting it with the tags #DeckTheLab and @Addgene. Christmas, Hanukkah, ugly...you name it, we want to see it. All photos tagged with #DeckTheLab throughout the month of December... -
Plasmids 101: A Brief History of Plasmids and an Improved eBook!
TypeBlog Post...for any extrachromsomal hereditary determinant”. His proposal, however, was basically ignored. A separate...chapters on genome engineering, cloning, and plasmid tags, addtional content added to the "Plasmids that Glow...Learn more about the history of plasmids and cloning: CSHL Meeting: Plasmids: History & Biology The Joshua... history of plasmids... -
Biosensor AAV Preps
TypeCollection...pAAV_hsyn_NES-his-CaMPARI2-WPRE-SV40 Syn CaMPARI2 his Constitutive 1 Schreiter 101061 pAAV_hsyn_NES-his-CaMPARI2...F391W-WPRE-SV40 Syn CaMPARI2-F391W his Constitutive 1 Schreiter 101064 pAAV_hsyn_NES-his-CaMPARI2-L398T-WPRE-SV40...L398T-WPRE-SV40 Syn CaMPARI2-L398T his Constitutive 1 Schreiter Calcium Sensor: jRGECO1 100852 pAAV.CAG.Flex.NES-jRGECO1a.WPRE.SV40...Sensors GRAB_NE nLight Oxytocin (OT) Sensors GRAB_OT Voltage Sensors Archon Voltron JEDI-2P ATP Sensors iATPSnFR2...-ChrimsonR-ST Syn jGCaMP8s none Cre-dependent 9 Histed 176749 AAV-Syn-H2B-jGCaMP8s-WPRE Syn jGCaMP8s none...GRAB_OTmut (control) none Constitutive 1, 9 Li Voltage Reporter: Archon 108422 pAAV-CAG-FLEX-Archon1-KGC-EGFP-ER2...KGC-GFP-ER2] Syn Archon1 EGFP Cre dependent 8 Boyden Voltage Reporter: Voltron 119036 pAAV-hsyn-flex-Voltron-ST... -
Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs
TypeBlog Post...and gram negative bacteria periplasm. Efforts to tag endogenous genes for proteins in these compartments...and Lindsey M. Costantini! Erik Lee Snapp received his PhD in Molecular Microbiology and Immunology from...from Oregon Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz...the Department of Anatomy and Structural Biology. His interests include the quality control of secretory...Munro, Sean, and H. R. Pelham. "Use of peptide tagging to detect proteins expressed from cloned genes:...Avaliable at Addgene Read Our Blog Post on Protein Tags Read Other Posts on Fluorescent Proteins Browse ...FP-fusion proteins functionally relative to the untagged protein of interest. To avoid the accumulation... -
Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR
TypeBlog Post... Replace the biotinalatable tag on dCas9 with a BioTAP-tag. The BioTAP-tag is a 69 aa long biotinylation... lentiviruses delivering 1) a BioTAP-tagged dCas9 and an eGFP tag, and 2) gRNAs Number of loci targeted...normally expressed in eukaryotic cells. Using this tag reduces the number of CAPTURE components that need...from three to two. Lentiviral delivery of BioTAP-tagged dCas9 and gRNAs. Using lentiviral delivery of the...system Stable cell line expressing 1) FLAG-biotin-tagged dCas9, 2) BirA, and 3) one or more gRNAs Two separate...conformation capture (3C) based methods, such as 5C or Hi-C (Han et al., 2018). Think of 3C methods as taking...outperformed other 3D chromatin methods such as ChIA-PET or Hi-C, by increasing both the number of unique chromatin... -
22 Hot Plasmid Technologies from 2014
TypeBlog Post...contain a variety of N- and C-terminal tags (including His, S-tag, OneStrep, CBP, Trx, GST, Halo, MBP, ...electron microscopy (EM) tag providing superior subcellular localization of tagged proteins compared to traditional...distributing his Designer Receptors Exclusively Activated by Designer Drugs (DREADD) plasmids on his own, UNC-Chapel...promoter, N-terminal tag, coding sequence of the gene of interest, C-terminal tag, plant terminator, and...of an isopeptide bond between 2 peptide tags, SpyTag and KTag. The group demonstrated the use of the SpyLigase... risk of off-target effects. Harald Janovjak and his team at the Institute of Science and Technology Austria...expression levels. APEX2 is a monomeric, 27 kD genetic tag which can be fused to proteins of interest for EM... -
In Vivo Biotinylation of Bacterial Fusion Proteins
TypeBlog Post...controlled easier than formation of GST dimers. Avi-Tag plasmid for improved biotinylation selectivity A ...fusion protein from a bacterial extract. The MBP tag can be removed using Xa protease. The carboxyl-terminus... pMal plasmid (NEB) thus creating the pMal-T-Avi-His/BirA plasmid now available through Addgene. We have...Octet, Pall ForteBio). Tips for using pMal-T-Avi-His/BirA to add biotin to your protein of interest Grow...amino acid) AviTag sequence in a protein and adds biotin to a lysine residue within the AviTag. However,...carboxyl-terminus of the fusion protein includes an AviTag and a 6X HisTag for affinity purification (see plasmid ...dramatically increased accessibility of the AviTag and HisTag, which were “buried” in a few of our proteins... -
SpyLigase Irreversibly Locks Peptides Together for Efficient Cell Capture
TypeBlog Post...join one peptide tag to another peptide tag. Addgene: What is the advantage of peptide tags for building ...locking two peptide tags together. SpyLigase overcomes limitations in the use of peptide tags, which often form...Howarth: Peptide tags can be very generic; you should be able to use the same peptide tag for all sorts ...the National Academy of Sciences in March, he and his colleagues took another important step forward by...Natl Acad Sci U S A. 2014 Mar 17. PubMed) Peptide tag forming a rapid covalent bond to a protein, through...cancer in particular. They recently introduced the SpyTag/SpyCatcher system, based on a protein isolated ...Howarth: It came from our work in developing the SpyTag/SpyCatcher system, to be able to grab on to proteins... -
When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells
TypeBlog Post...solution or something in between, you are using a tag with a serious drawback. The standard EGFP plasmid... of a POI. As an investigator, you want a fusion tag to be inert, to not produce artifacts in your experiments...guest blogger Erik L. Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology and Immunology from...from Oregon Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz...the Department of Anatomy and Structural Biology. His interests include the quality control of secretory...oligomers such as DsRed or high affinity FPs such as TagRFP. Problem 3 can be solved, in most cases, by using... -
Molecular Biology Reference
TypeGuide...prokaryotes. Common Epitope Tags Tag Amino Acid Sequence FLAG DYKDDDDK HA YPYDVPDYA His HHHHHH Myc EQKLISEEDL...nucleotides. Epitope tags on the other hand are commonly used in molecular cloning to tag a gene within a ...E GAA, GAG Glycine Gly G GGU, GGC, GGA, GGG Histidine His H CAU, CAC Isoleucine Ile I AUU, AUC, AUA Leucine...random incorporation of modified, fluorescently tagged bases onto the growing DNA strand in addition to...T, C, or G nucleotide. The 4 standard bases are tagged with a different fluorophore so they can be distinguished...when the polymerase incorporates a fluorescently tagged nucleotide. Because these special bases do not ..., the reaction is halted once the fluorescently tagged base is incorporated. Sanger sequencing requires... -
Even more elegant: Single injection CRISPR/Cas9 in C. elegans
TypeBlog Post...illustrating the steps to fluorescently tag a C. elegans protein. The his-72 locus is shown as an example. Used...time. Creating a modular, selection-based gene tagging approach Multiple features of C. elegans have made... between a fluorescent protein (FP) and a 3xFLAG tag (see construct schematic below.) After self-excision...intron, and only the fluorescent protein and 3xFLAG-tag sequences are inserted into the genome. Figure...template plasmid to create fluorescent and 3xFLAG-tag fusion proteins. The repair template and Cas9-sgRNA...little hands-on labor and could facilitate the tagging of every protein in the C. elegans genome. Although... gene tagging... -
Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers
TypeBlog Post...system for protein tagging. We spoke with Mendenhall and Myers about their new CRISPR tagging system, which...www.addgene.org/crispr/tagging. Addgene: What motivated you to adapt CRISPR for use in protein tagging? Myers: Our...epitope tag onto the transcription factors. By the way, you can actually use this for tagging any protein...Alabama in Huntsville explains it, a major goal in his laboratory is to understand the function of the non-coding...factors. Our groups got together to adapt CRISPR for tagging experiments and it’s already looking like it’s ...correctly, the second plasmid inserts a small epitope tag called FLAG at the end of the transcription factor...you can select the population of cells with the tagged transcription factor. The FLAG antibody has been... -
Tetbow: Bright Multicolor Labeling for Neuronal Tracing
TypeBlog Post... with containing chemical tags such as the SNAP-tag, Halo-tag, and CLIP-tag (Kohl et al., 2014) (Addgene...Takeshi Imai is a professor at Kyushu University. His lab has been studying functional and anatomical aspects... et al. "Ultrafast tissue staining with chemical tags." Proceedings of the National Academy of Sciences111.36... circuits with fluorescent proteins and chemical tags." eLife 7 (2018): e40350. PubMed PMID: 30454553.... -
Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification
TypeBlog Post...have two options: Option #1: Affinity tag purification You tag YFP and use an affinity column for purification...protein. OptoNBs were also tagged with an infrared fluorescent protein tag which let the team detect its... used nickel-coated agarose beads coated with his-tagged OptoNBs. OptoNB coated beads were then imaged... be tagged, which limits these approaches to use with recombinant proteins. Purifying an untagged protein...Opto-Nanobodies (OptoNBs) purification You skip adding a tag to YFP and instead use OptoNBs. You fill a column...protein of interest in response to light allows for tagless protein purification as well as reversibly regulation...domain and a nanobody. AsLOV2 is a light-oxygen-voltage sensing domain from the oat plant (Avena sativa... -
RANbodies: Reporter Nanobody Fusions
TypeBlog Post...easy to harvest from cell culture media. Their His tags allow for further purification, if desired, but...RANbody is detectable by two different tags: a reporter and an HA tag. The reporter comes in three varieties...Spaghetti Monsters are highly antigenic tags that have 10 HA or 10 MYC tags built into a GFP scaffold. Some GFP... secondary antibody staining HA Tagged GFP, RFP, H2A2B Myc Tagged RFP, H2A2B Chicken IgY (Y) Labeled...lot-to-lot variability in activity. No Genetic Tagging - Since antibodies are produced in animals instead... via recombinant DNA, they can’t be genetically tagged and instead are conjugated to reporters (i.e. fluorescent...which are discussed in more detail below. The HA tag can be stained for with an anti-HA antibody and serves... -
CRISPR 101: Ribonucleoprotein (RNP) Delivery
TypeBlog Post...express and purify His-tagged Cas9 from E. coli using a plasmid such as pET-28b-Cas9-His from Alex Schier...such as visualizing genomic loci using fluorophore tagged dCas9 may require the use of plasmid or viral-mediated...genomic deletions. What are the advantages of Cas9 RNPs? There are many advantages to using RNPs for CRISPR ... Delivering Cas9-gRNA ribonucleoproteins An advantage of performing CRISPR experiments using RNPs is... can be used to deliver a Cas9-gRNA RNP. This advantage has allowed scientists to perform in vitro and...transfection (B in the figure above), are in the early stages of development. David Liu’s Lab has demonstrated... -
New Podcast Segment: Hot Plasmids
TypeBlog Post...from John Chodera's Lab - Get high expression of His-tagged human kinases in bactera. Listen to Our Segment... -
The Breast Cancer Microenvironment: A Tumor’s Backstage Team
TypeBlog Post...Allinen et al., 2004). In fact, Sanford H. Barsky and his team tested the influence myoepithelial cells had...touring musician needs? A backstage team. Meet the microenvironment, a tumor’s backstage team. Microenvironments... seem to take center stage in cancer development, the microenvironment backstage play just as important...breast tumorigenesis: Friend or foe?." Histology and histopathology (2018): 18021-18021. PubMed PMID: 29978449...microenvironment’s physiology is deeply connected to these 3 stages and contains multiple cell types: fibroblasts, ... -
Better Dyeing Through Chemistry & Small Molecule Fluorophores
TypeBlog Post...concept include: Self-labeling tags (e.g., SNAP-tag, HaloTag, TMP-tag) – These widely used systems consist...strategies have two parts: (1) a genetically encoded “tag” expressed as a fusion with your favorite protein...fluorophore-contianing “ligand” that binds to the tag. Like most good ideas in biological imaging, the ...genetically encoded tetracysteine (Cys4) peptide tag could be used to label proteins in cells (Figure ...consist of a genetically encoded enzyme variant tag that reacts specifically and irreversibly with a small... attachment of a fluorophore ligand to a peptide tag (Figure 1c). Click chemistry (e.g., transcyclooctene–tetrazine...paclitaxel. Unlike other systems, no genetically encoded tag is required as the small-molecule binding motif targets... -
Advice for Scientists Starting a Lab
TypeBlog Post... lead his team as a scientists and a friend. George Church runs his lab fairly similarly to his advisor...the majority of his time in the lab. As time went on and he was spending more time in his office, he gradually... always “off flying his plane to exotic places while occasionally emailing us his worldly advice”, the...package that included a postdoc and a PhD student in his first year. “It was very important to me to be with...adviser did? Tom Ellis merges the influence from his two PhD advisers who were “chalk-and-cheese”. While... build a strong, collaborative team. And take advantage of the many resources available from your institution... -
Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools
TypeBlog Post...labels proximal proteins with biotin as an affinity tag. This ‘Caspex’ construct is guided by a pre-defined...segment Similarly to the Carr lab, Erik Sontheimer and his group from the University of Massachusetts Medical...biotinylation at genomic elements by restricted spatial tagging), to confirm the composition of proteins found ...antibodies found in camelids. They possess many advantages over conventional antibodies: they are small ... -
Hot Plasmids: FRET-Based Biosensors
TypeBlog Post...he hopes others will find his tools useful, including his zinc sensor and his genetically encoded magnesium...on the move, such as T lymphocytes. Griesbeck and his colleagues began by isolating a calcium-binding protein...Ratiometric imaging for beginners Griesbeck says his calcium indicators can be used in mouse, Drosophila...Griesbeck also points out that the approach he and his colleagues took to optimize sensitivities can now...other published FRET-based sensors. For example, his team is now applying it to improve upon an indicator...than calcium, requiring greater sensitivity. He and his colleagues came up with a solution: they devised ... adds that this color change has a particular advantage for observing events taking place inside cells...