We narrowed to 400 results for: pho;
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TypeCollection...generate the phosphosites with serine instead of phosphoserine (i.e. a “phosphorylatable” phosphosite library... serine phosphorylation ( Rinehart Human Serine Phosphopeptide Library ). The phosphorylation sites (“...Reagents Rinehart Lab Phosphoprotein Reagents Protein phosphorylation is one of the most abundant forms of...questions surrounding serine phosphorylation by adding this phosphorylated amino acid to the genetic code...or multiply phosphorylated proteins (Pirman et al., Nat Comm 2015). Improved phosphoprotein production ...(“phosphosites”) consist of a central phosphoserine site flanked by 15 amino acids on either side occurring... site-specific incorporation of phosphoserine in >36,000 phosphosites by tandem mass spectrometry (Barber...
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Using Phosphoserine to Study Protein Phosphorylation
TypeBlog Post...SDS-PAGE gels containing Phos-Tag, a small molecule that binds specifically to phosphate moieties and promotes... al. "Detection of phosphorylation on large proteins by Western blotting using Phos-tag containing gel...MEK1 phosphomimetic residues in the above study with phosphoserine (to create a pSer/phosphomimetic dual...shift in phosphorylated, but not non-phosphorylated proteins (8). Quantifying the ratio of phospho- and non-phospho-bands...of the dephosphorylation reaction can be tracked through the comparison of phospho:non-phospho species...modified through phosphorylation (1). However, the expansion of studies on protein phosphorylation in an array...suggest that phosphorylation is far more prevalent than previously appreciated. PhosphoSitePlus, one of the... -
Synthetic Photobiology: Optogenetics for E. coli
TypeBlog Post...or dark conditions, the chromophore activates the RR (OmpR) via phosphorylation, promoting transcription...histidine kinase/photosensor senses various light inputs, and an obligate chromophore allows the system...dark conditions, the chromophore activates the response regulator via phosphorylation, promoting transcription...in E. coli easier and more robust. Synthetic photobiology: why use light? In synthetic biology and bioengineering...TCSs). TCSs consist of a histidine kinase that phosphorylates a response regulator (RR). TCS-induced responses...two-component system (TCS). Levskaya et al. fused a photosensory domain from cyanobacteria to a common E. coli...hybrid construct senses light, and an obligate chromophore allows the system to respond to various light... -
Fluorescent Proteins 101: Aptamer Fluorophores
TypeBlog Post...state contains a fluorophore and a quencher in close proximity. For instance, the fluorophore and quencher...of aptamers means they can be used with or as fluorophores in several different ways. Researchers have ...fluorescent in situ hybridization (FISH) where fluorophore-linked antibodies are used to visualize RNA. ...removing the quencher from the proximity of the fluorophore and subsequently leading to a positive signal...signal. The same effect can be achieved for a fluorophore that relies on dimerization for activation, only...used as powerful biosensors, the bound protein fluorophores and quenchers are still relatively bulky. One...fluor. Transcribing and translating a protein fluorophore also takes time - bypassing translation altogether... -
Lighting Up Cell Signaling with Photoswitchable Kinases
TypeBlog Post...dimerizing photoswitchable Dronpa (dron= a ninja term for vanishing, and pa= photoactivation) domains on... Figure 1: Photoswitchable MEK1 (psMEK1). Is it possible to create other photoswitchable kinases? The...Signal transduction pathways are a lot like cell phone networks. Protein kinases deliver messages to the...-specifically activated and, like the game of telephone, often the signal must be transmitted by intermediate... lab present a generic design for creating photoswitchable kinases and use MEK1 as a model kinase. In ...site of a constitutively active MEK1. This photoswitchable MEK1 is called psMEK1. When exposed to 500 ...psMEK1 also worked for creating three other photoswitchable kinases: psMEK2, psRaf1, and psCDK5. MEK2 is... -
Seeing Red: Simple GFP Photoconversion
TypeBlog Post...biological research. Photoconvertible fluorescent proteins Newly developed photoconvertible FPs like PsMOrange...Applications and limitations of GFP photoconversion One concern with photoconverting GFP is the expression level...and efficiently photoconverted in plant, Drosophila, and mammalian cells. Photoconverted GFP is compatible...Davidson MW, Looger LL. A bright and photostable photoconvertible fluorescent protein. Nat Methods. 2009...accidental discovery that laser treatment can photoconvert GFP from green to RED! This simple technique...time-consuming to repeat the process with these new photoconvertible proteins. With Sattarzadeh et al.’s discovery...discovery that GFP itself is photoconvertible, new studies of spatiotemporal dynamics can be undertaken using... -
Choosing the B(right)est Fluorescent Protein: Photostability
TypeBlog Post...emitted photons before they are destroyed. Factors that affect photostability The photobleaching rate of...will discuss how to select a photostable fluorescent protein. Photobleaching is the irreversible destruction...destruction of a fluorophore under the influence of light. Any fluorescent molecule will photobleach at some point...have fluorescent proteins that are photostable. On top of photobleaching, fluorescent proteins may display...likelihood that a fluorophore will bleach. This is, however, not the full story. The photobleaching rate does ... 2-fold does not reduce photobleaching by half. How exactly the photobleaching rate depends on power is...to compare the photostability of different fluorescent proteins? Measuring photostability Since it is clear... -
Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins
TypeBlog Post...illuminated with UV-violet light (~400 nm), the chromophore photoconverts predominantly to the anionic form, causing...presented first, it causes irreversible photoconversion of the chromophore from the neutral to the anionic form...mEosFP, can form it’s chromophore at a more comfortable 37°C (5). Dendra2 photoconverts from green to red ...reversible PA-FPs (also known as photoswitchers) can be photoswitched from a dark state to a bright fluorescent... for vanishing, and pa, for photoactivation) (6). Dronpa is photoactivated by blue wavelengths of light...consider, such as brightness, photostability, pH stability, chromophore maturation rate at 37 °C, turnover...-FPs offer a reduced potential for photobleaching and phototoxicity that standard FPs might induce though... -
Special Delivery: Fluorophore Targeting for FRET Studies
TypeBlog Post...biophysical interaction between two fluorophores. When the first fluorophore (the donor) is excited, it can...acids/lipids of interest are fluorophore labeled and that these fluorophores can undergo FRET when they ...targeting a FRET fluorophore to a biomolecule is a bit like a Fedex delivery. Your fluorophore package needs... energy directly via resonance to the second fluorophore (the acceptor) if the two probes are close together...physical distance between the donor/acceptor fluorophores (Stryer, 1978), a measurement that is unaffected...label your biomolecule of interest with FRET fluorophores? When is absolute labeling specificity needed...measure it). Molecular mailbox: How to get your fluorophore delivered to the right place? FRET experiments... -
Photosensitizer Induced Cell Ablation with FAP-TAP MG-2I-dL5**
TypeBlog Post...technique! What is a photosensitizer? First, for a few definitions. A photosensitizer is a dye or protein... higher photostability than KillerRed, with KillerRed reaching greater than 75% photobleaching after just...system? Turns out that with light-inducible photosensitizers and a quick zap of the proper color light,...light, you can do just that. Photosensitizing dyes and proteins have been around for awhile (check out this...Bruchez and Tsang labs recently developed a photosensitizer composed of the protein complex complex and... oxygen species (ROS). The ROS produced by photosensitizers inactivate target proteins or cells while ...having little effect on neighboring cells. The photosensitizer used in this work is composed of two parts:... -
Better Dyeing Through Chemistry & Small Molecule Fluorophores
TypeBlog Post...called “the photon budget”: each sample has a limited number of fluorophores and each fluorophore can emit...the photon budget and pushing the frontiers of fluorescence microscopy often requires more photons. For...cells can decrease the number of fluorophores, compromising the photon budget. Likewise, the switch from...desperate to extract a few more photons from a sample. Chemical fluorophores can be substantially brighter...more photostable than fluorescent proteins, providing a straightforward way to improve the photon budget...rhodamine fluorophores, such as the Alexa Fluor and ATTO dyes, with improved brightness, photostability, and... of photons for your imaging experiment. Innovative labeling strategies and improved fluorophores are ... -
Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins
TypeBlog Post...Listen to the photoswitchable protein podcast segment Reversible control of photoswitchable proteins, which...., 2018). They accomplished this by inserting a photo-sensitive LOV2 domain from A. sativa into a loop...ratio, high sensitivity, responds quickly, is photostable, and is compatible with optogenetic control. ... Discovery of new protein binder tools for photoswitchable proteins cPYP and AsLOV2 Article contributed...temporally control cellular processes in vivo. Many photoactive proteins exist in nature, but developing them... either the light- and dark- states of the photoswitchable cPYP, which changes in conformation in response...bind to only the light- or dark- state of the photoswitchable proteins offers a powerful approach to control... -
Immunology Research Plasmids and Resources
TypeCollection...PLCG2 phospholipase C, gamma 2 (phosphatidylinositol-specific) - PPP3CA protein phosphatase 3 (formerly...IP1, IP2, IPD2, NEMO INPP5D inositol polyphosphate-5-phosphatase, 145kDa MGC104855, MGC142140, MGC142142...FLJ26625, JTK8 MALT1 mucosa associated lymphoid tissue lymphoma translocation gene 1 DKFZp434L132, MLT... PIK3CD phosphoinositide-3-kinase, catalytic, delta polypeptide p110D PIK3CG phosphoinositide-3-kinase...and cytoplasmic domain, (semaphorin) 6D FLJ11598, KIAA1479 SEMA7A semaphorin 7A, GPI membrane anchor (...BMP1 bone morphogenetic protein 1 FLJ44432, PCOLC, PCP, TLD, pCP-2 BMP10 bone morphogenetic protein 10...MGC126783 BMP15 bone morphogenetic protein 15 GDF9B, ODG2, POF4 BMP2 bone morphogenetic protein 2 BMP2A BMP3... -
Genetic Code Expansion
TypeCollection... Expansion Accessible Using Phosphoserine to Study Protein Phosphorylation Rinehart Lab reagents tRNA/... mutated, serB deleted for phosphoserine GCE. For expressing phosphoserine-containing proteins using genetic... pMAH2-CageCys leucyl-tRNA-synthetase E. coli photocaged cysteine Mammalian TAG Huiwang Ai 73544 pEvol-pAcFRS...91705 pSupAR-MbPylRS(DiZPK) PylRS M. barkeri photocrosslinkers DiZPK, DiZSeK, or DiZHSeC Bacterial TAG P....91706 pCMV-MbPylRS(DiZPK) PylRS M. barkeri photocrosslinkers DiZPK, DiZSeK, or DiZHSeC Mammalian TAG P.... PylT_FLAG-AbKRS-chIPYE chimeric PylRS Chimeric photo-crosslinking-lysine Mammalian TAG Simon Elsaesser...140022 pAS_4xMma PylT_FLAG-PcKRS PcKRS M. barkeri photocaged-lysine Mammalian TAG Simon Elsaesser 140023 pAS... -
Antibody Guide
TypeCollection... curve with known amounts of protein. Fluorophores Fluorophores are light-sensitive molecules which emit...IF uses fluorophore color to differentiate between the targets of interest, if your fluorophore is bound...the fluorophores used is important. It is best to inversely pair the brightness of the fluorophore with...If a bright fluorophore is on an abundant target, it obscures fainter signals. Fluorophores can emit signals...storing your antibody: Antibodies conjugated to fluorophores should never be stored below 4 °C. Certain isotype...molecules are horseradish peroxidase (HRP) and fluorophores. Horseradish peroxidase (HRP) HRP performs a...specific wavelength. While more expensive than HRP, fluorophores are available in a range of colors activated... -
Fluorescent Protein Guide: Biosensors
TypeCollection...Webb Phosphate FRET-based phosphate sensor (FLIPPi) A novel analytical method for in vivo phosphate tracking...Ca(2+) indicator with enhanced two-photon absorption. Neurophotonics. 2024 Apr;11(2):024207. doi: 10.1117...29;7. pii: 32638. Kai Johnsson Phosphate Scaffold for inorganic phosphate biosensor Development of a Reagentless.... Eric Schreiter Calcium AAV expression of photoconvertible fluorescent protein-based calcium integrator...calcium sensor with exceptional brightness, photostability, and multiplexing capabilities. bioRxiv. 2025... jYCaMP: an optimized calcium indicator for two-photon imaging at fiber laser wavelengths. Nat Methods...localized zinc sensor ZIBG2 Genetically Encoded, Photostable Indicators to Image Dynamic Zn(2+) Secretion ... -
Ras Pathway
TypeCollection... Protein phosphatase 1 catalytic subunit alpha PREX2 Phosphatidylinositol-3,4,5-trisphosphate-dependent...receptor PDPK1 3-phosphoinositide dependent protein kinase 1 PEBP1 Phosphatidylethanolamine binding protein...PIK3R5 PIK3R6 Also known as PI3K; Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic and regulatory...state, regulated by the addition and removal of a phosphate group. Under normal physiological conditions GTPases...thymoma viral oncogene homolog 3 ALK Anaplastic lymphoma receptor tyrosine kinase ARHGAP35 Rho GTPase activating...DUSP2 DUSP3 DUSP4 DUSP5 DUSP6 Dual specificity phosphatase E2F E2F1 E2F2 E2F3 E2F transcription factor ECT2...G1, G2, G3: non-catalytic subunit gamma PTEN Phosphatase and tensin homolog RAC RAC1 RAC2 RAC3 Ras-related... -
Optogenetics AAV Preps
TypeCollection... Dlx EF1a/nEF Synapsin E2 regulatory element Fluorophore GFP Red-wavelength fluorescent protein Yellow-wavelength...pAAV-CaMKIIa-ChRmine-mScarlet-Kv2.1-WPRE CamKII ChRmine (high-photocurrent, red-shifted) mScarlet Constitutive 8 Deisseroth...pAAV-hSyn-ChRmine-mScarlet-Kv2.1-WPRE Syn ChRmine (high-photocurrent, red-shifted) mScarlet Constitutive 8 Deisseroth...pAAV-Ef1a-DIO-ChRmine-mScarlet-WPRE EF1a ChRmine (high-photocurrent, red-shifted) mScarlet Cre dependent 1, 5 Deisseroth... 6m-p2A-ChRmine-Kv2.1-WPRE Syn ChRmine (high-photocurrent, red-shifted) GCaMP6m Constitutive 8 Deisseroth... pAAV-nEF-ChRmine-mScarlet nEF ChRmine (high-photocurrent, red-shifted) mScarlet Constitutive 8 Deisseroth...pAAV-nEF-Con/Fon-ChRmine-oScarlet nEF ChRmine (high-photocurrent, red-shifted) oScarlet Cre and Flp dependent... -
Fluorescent Protein Guide: In Vivo Imaging
TypeCollection... multicolor imaging. Photo-activatable iRFPs can be ‘turned on’ by non-phototoxic far-red light and are...690/717 (after photoactivation) 3.2 pPAiRFP1-N1 PAiRFP2 692/719 (after photoactivation) 3.0 pPAiRFP2-N1... imaging using far red, near-infrared, and photoactivatable fluorescent proteins. Plasmid...imaging are derived from bacterial phytochrome photoreceptors (BphPs). Spectrally distinct permanently fluorescent...The following far red , near-infrared , and photoactivatable fluorescent empty backbone plasmids are ideal...piRFP720-N1 iSplit 690/713 5.3 pPAS-E and pK-GAFm Photoactivatable Protein Excitation/Emission Brightness Find... -
Bacterial Expression Systems
TypeCollection... Large Stokes Shift , Photoactivatable , Photoconvertible , and Photoswitchable fluorescent proteins Various...monitor in live cells (beware of artifacts!), photoactivatable tags for super resolution imaging, epitope...20341 mEos2 (fluorescent protein) Loren Looger Photoactivable Fluorescent Protein Expression: contains mEos2...mEos2 with an N-terminal His tag. Photoactivable fluorescent proteins like this one can be used in super...protein, MS2. Can be used in combination with a fluorophore-tagged MS2 (such as that found in pZS*12-MS2-... to dsRED. Can be used in combination with a fluorophore-tagged MS2 (such as that found in pZS*12-MS2-...enhances the FRET signal from the CFP and YFP fluorophores linked by an arabinose binding protein (AraF... -
Luciferase Plasmid Collection
TypeCollection... of photons. The robust signal, sensitivity, wide dynamic range, and resistance to photobleaching has ... tag with a fluorophore shifts the emission maximum of NanoLuc to that of the fluorophore. Conformationally... sensor created by fusing NanoLuc® and Venus fluorophore to Troponin C Ca 2+ binding domain. Nano-lanterns...-Leu linker to the circularly permuted Venus fluorophore. Learn more about pHlash sensor in our 28 Hot...Firefly Creation of N-terminal Firefly fusions in Drosopholia Michael Rosbash 128046 pGL3basic luciferase and...Insertion of 5' promoter/enchancer regions in Drosopholia Herman Wijnen 126033 pTETRIS-cargo Firefly For... -
Cre-lox system
TypeCollection...pAAV-CIBN-CreC photo-activatable cre recombinase CAG AAV Bruchas 75268 pAAV-CRY2-CreN photo-activatable ... Cre and gRNAs cTnT AAV Pu 87694 pCAG-ERT2-PhoCl-Cre-PhoCl-ERT2 light-activated Cre cTnT Mammalian Campbell...Cepko 13780 pNrl-Cre Cre-Myc, Expressed in rod photorecetor cells in the retina Nrl Mammalian Cepko 13962...pAAV-Syn-iCre iCre AAV Ritola 122960 pcDNA3.1_PA-Cre photo-activatable Cre recombinase CMV Mammalian Sato 122961...pBbE5a-Opto-Cre-Vvd Cre recombinase split with Vivid photodimers lacUV5 Bacterial Dunlop 135217 pDEST mfap4:icre-p2a-tomato...pAAV-EF1a-NCreV N-terminal Cre component split with Vivid photodimers EF-1 alpha AAV Zeng 140132 pAAV-EF1a-CCreV C-...C-terminal Cre component split with Vivid photodimers EF-1 alpha AAV Zeng 140135 pAAV-EF1a-iCreV Split... -
Recombinases AAV Preps
TypeCollection...these molecular tools. Cre AAV ID Name Promoter Fluorophore/Tag Serotype(s) PI 105551 pENN.AAV.CamKII.HI....AAV.TBG.PI.Cre.rBGe TBG none 8 Wilson Dre AAV ID Name Promoter Fluorophore Serotype(s) PI 50363 AAV phSyn1(S)-DreO-bGHpA...Syn none 5, rg* Zeng Flpo AAV ID Name Promoter Fluorophore Serotype(s) PI 51669 AAV phSyn1(S)-FlpO-bGHpA...Light-Inducible Recombinases ID Name Promoter Fluorophore Serotype(s) PI 140135 pAAV-EF1a-iCreV EF1a none...none 1, PHPeB Zeng VCre AAV ID Name Promoter Fluorophore/Tag Serotype(s) PI 55638 pAAV-EF1a-vCre EF1a ... -
Zhang Lab CRISPR Page
TypeCollection...to enable the rapid ligation of annealed and phosphorylated oligos designed based on the target site sequence... -
Fluorescent Protein Guide: Empty Backbones
TypeCollection...Near-Infrared Long Stokes Shift Photoactivatable Photoconvertible Photoswitchable Timers Blue/UV Protein Excitation...LSSmKate2-C1 - Mammalian Expression Jump to Top Photoactivatable (e.g. off to on) Excitation and Emission wavelengths...HisB-PAiRFP2 - Bacterial Expression Jump to Top Photoconvertible (e.g. green to red) Protein Excitation (nm...pBAD-PSmOrange2 - Bacterial Expression Jump to Top Photoswitchable (e.g. off to on to off) Protein Excitation ... -
mTOR Pathway
TypeCollection...PIK3R2 PIK3R3 PIK3R4 PIK3R5 PIK3R6 Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic and regulatory...AKT1S1; AKT1 substrate 1 (proline rich) PTEN Phosphatase and tensin homolog Rag RRAGA RRAGB RRAGC RRAGD...Protor Also known as PRR5; proline rich 5 PTEN Phosphatase and tensin homolog Rictor RPTOR independent companion... -
p53 Pathway
TypeCollection...deprivation. p53 is activated through both phosphorylation and acetylation, which disrupt its interaction...ligase MDMX p53 regulator; also known as MDM4 Noxa Phorbol-12-myristate-13-acetate-induced protein 1; also... 1 (RCHY1); E3 ubiquitin protein ligase PTEN Phosphatase and tensin homolog PUMA BCL2 binding component... -
Genomic Deletions in Mammalian Cell Lines
TypeCollection...2.1 may be omitted. CRISPR Cloning Anneal and phosphorylate oligos. Resuspend oligos at a concentration ...Polynucleotide Kinase (PNK) (10,000 U/ml). NOTE: Phosphorylated oligos may be ordered instead. For this approach... -
Brzezinski Lab CRISPR Collection
TypeCollection...followed by a quick ligation of annealed and phosphorylated oligonucleotides containing the guide target...Prdm1 and Vsx2 enhancers in the retina alters photoreceptor and bipolar cell fate specification, yet differs... -
Antibody Production
TypeCollection...are initially titered by spectrophotometry using a NanoDrop spectrophotometer. If needed, the sample is...centrifugal columns. The final formulation buffer is phosphate-buffered saline containing 1 mM sodium azide. ... -
Zebrafish Plasmid Collection
TypeCollection...transposons, Cre-mediated site-specific recombination, morpholinos, zinc finger nucleases, TALENS, and CRISPR/Cas9...of gene function in zebrafish - Julian Lewis Lab PhOTO Zebrafish: A Transgenic Resource for In Vivo Lineage...consisting of plasmids with four promoters, six fluorophores, and empty vectors designed to streamline the... -
Brain Initiative Collection
TypeCollection...Cation channelrhodopsin ChrimsonR fused to mRuby2 fluorophore and targeted to the neuronal soma and proximal...parapinopsin driven by the CAG promoter. Used for photoswitchable control of inhibitory GPCR signaling cascades...parapinopsin driven by the Ef1a promoter. Used for photoswitchable control of inhibitory GPCR signaling cascades... -
Trimmer Lab NeuroMab Collection
TypeCollection... Ca2+ channel phosphorylation site pS1928 [L132/18R] Cav1.2 Ca2+ channel phosphorylation site pS1928 Rat...Anti-Kv1.2 pS440/pS441 phosphospecific [L64/32] Kv1.2 pS440/pS441 phosphospecific Human Mouse IgG2a 190291... -
CRISPR Guide
TypeCollection...SpCas9-HF1 - disrupt Cas9’s interactions with DNA phosphate backbone HypaCas9 - increase Cas9 proofreading...fusing the anti-CRISPR protein AcrIIA4 with the photosensitive LOV2 domain . Several labs have also controlled...: Epigenetics Visualizing Genomic Loci Using Fluorophores CRISPR imaging has numerous advantages, including...11: Overview of visualizing genomic loci with fluorophores using CRISPR Browse Plasmids: Visualize Purify...Yoshimi, K., Okuzaki, Y., Gee, P., Kunihiro, Y., Sonpho, E., Xu, H., Sasakawa, N., Naito, Y., Nakada, S... -
Michael Davidson Fluorescent Protein Collection
TypeCollection...mEmeralds Reds Blues Cyans Oranges Greens Yellows Photoactivatables Empty Backbones Plasmid Description Industry...Industry Yellows Plasmid Description Industry Photoactivatables Plasmid Description Industry Do you have suggestions... -
Depositor Collections
TypeCollection...Reagents for improved expression of recombinant phosphoproteins - Rinehart FreeGenes Project Open Enzyme Collection... -
Synthetic Biology - Overview
TypeCollection...Optogenetics Pertz Lab cpFRET Kit Rinehart Recombinant Phosphoprotein Kit S. cerevisiae Advanced Gateway Destination... -
Worm Expression Resources
TypeCollection...uses purified Cas9, such as that expressed from pHO4d-Cas9 from Michael Nonet. Protocol can be found at...and GFP fusion vectors. C. elegans optimized fluorophores - Andrew Fire Lab. Available with and without... -
Allen Institute for Cell Science Plasmid Collection
TypeCollection...Sialyltransferase 1 Golgi 109122 NPM1-mEGFP AICSDP-50 mEGFP Nucleophosmin Nucleolus (granular component) 109120 GJA1-mEGFP...z-disks 124608 NPM1-mTagRFP-T AICSDP-69 mTagRFP-T Nucleophosmin Nucleolus (granular component) 133963 UBTF-HaloTag... -
Kazuhiro Oka Lentiviral Vectors
TypeCollection...72268 xpress gene of interest from the PGK (phosphoglycerate kinase I) promoter pCDH-CMV4 72284 Express... -
Fluorescent Protein Guide: FRET
TypeCollection...non-radiatively transferred from an excited donor fluorophore to an acceptor. Since the transfer of energy...energy does not occur by emission of a photon, the acceptor molecule is not required to be fluorescent. A... -
Control AAV Preps
TypeCollection...EF1a/nEF GFAP and variants Synapsin TBG Other Fluorophore/Tag Green Red None Other Cre-dependent color ...non-cre-dependent) Clear Filters ID Name Promoter Fluorophore/Tag Activity Serotype PI 37825 AAV-CAG-GFP CAG... -
Viral Production
TypeCollection...preparations are subjected to polyacrylamide gel electrophoresis (PAGE) followed by silver staining or SYPRO...purified and non-purified AAV subjected to gel electrophoresis. Viral capsid proteins VP1, VP2, and VP3 are... -
Plan Your Experiment
TypeCollection...gRNA Presence of a selectable marker (drug or fluorophore) Delivery method Expression System Components...identify DNA fragments. PCR amplification and gel electrophoresis (for HDR or NHEJ): For large deletions or insertions... -
Deisseroth INTRSECT Collection
TypeCollection...configuration have been developed, including additional fluorophores, excitatory and inhibitory opsins , genetically-encoded...H134R)-eYFP Vcre No Dual recombinase-dependent: Fluorophores Addgene ID Plasmid Logic Sites and Mutations... -
Plasmids for Stem Cell Research
TypeCollection... Conversion of Adult Mouse Liver Cells and B Lymphocytes to Neural Stem Cells. Cell Reports. 2014 Dec ...Underlies Lineage Conversion into Functional Trophoblast Stem-like Cells. Cell Stem Cell. 2015 Nov 5;17... -
Antibodies 101: Designing Your First Flow Panel
TypeBlog Post...wavelengths of emitted photons are higher than those of absorbed photons. When photons excite electrons of... you'll need to pick a fluorophore for each target and ensure the fluorophores, together, are able to ... about 380–700 nm. Photons of lower wavelengths are higher in energy, while photons of higher wavelengths...whereby a photon is emitted. This step is called emission. During excitation, some of the photon's energy...the energy of an emitted photon is lower than the energy of the absorbed photon. Correspondingly, wavelengths...outlining a few commonly used fluorophores and their emission spectra. Some fluorophores carry their emission ...and RB545. Fluorophores can also come in combination of two conjugated single fluorophores, like APC-Cy7... -
Optogenetics + CRISPR, Using Light to Control Genome Editing
TypeBlog Post...design utilized Magnet photoswitchable proteins derived from the fungal photoreceptor, Vivid (VVD, N. crassa...split fusions and generate a photoactivatable nickase and a photoactivatable repressor (dCas9). The activity...Off Genome Editing Single-chain Cas9 photoswitches Photoactivation of Cas9 can also be achieved using single-chain...chemistry to photocage CRISPRs The aforementioned techniques each employed a photoactive strategy which...Comparison of photoactivatable CRISPR strategies Publication System nickname Photoactivatable moiety Cas9...et al., 2016 CRISPR-plus Photocleavable ssDNA oligonucleotides Cas9, photocaged-gRNA Gene editing Manna...optogenetic techniques to develop a variety of photoactivatable CRISPR tools. These tools allow scientists... -
Measuring Kinase Activity at the Single-Cell Level with Kinase Translocation Reporters (KTRs)
TypeBlog Post...proteins predicted to be phosphorylated on at least one site. Phosphorylation is particularly important...predict. Slower Dephosphorylation Rates: FRET-based reporters can have slower dephosphorylation rates than ...by FRET reporters having phosphorylated conformations that are less phosphatase-accessible. Kinase Translocation... signal (NES), both which are phosphorylatable. Prior to phosphorylation, the KTR localizes to the nucleus...biosensors Learn about using phosphoserine to study protein phosphorylation Additional Resources on Addgene.org...specific spectral properties that limit which fluorophores can be multiplexed together. Challenging to...nucleus, but following phosphorylation bNLS activity is suppressed and NES activity is enhanced, leading... -
Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins
TypeBlog Post...fluorescent proteins or dyes. 3. Two-photon microscopy Two-photon excitation microscopy is a fluorescence...decypher cellular dynamics in lymphoid organs. In lymph nodes, for example, two-photon microscopy allowed scientists...worthwhile to test the photostability of your fluorescent protein when using 2-photon microscopy as this ... is advantageous when compared to photoactivable or photoconvertible proteins, as it enables tracking ...mobility with movement from the bone marrow and other lymphoid organs to specific tissues where they can scan... good tools to directly follow immune cells in lymphoid organs. The development of cell sorters together...several labs were able to decipher the journey of lymphocytes from the bone marrow to the thymus where they...