We narrowed to 883 results for: prot

... 100 kD proteins or 20, 40, 60, and 80 kD proteins. Plasmids containing 150 or 250 kD proteins are also...expressed proteins run in separate lanes. B: individually expressed and coexpressed proteins run as ladders...for multiplexed gene perturbation The Penn State Protein Ladder Engineered miniature CRISPR-Cas system for...to clone new constructs to label your favorite protein! While the target only requires a small epitope...doi: 10.1038/s41587-020-0600-6. The Penn State Protein Ladder  by: Rachel Leeson This simple, inexpensive...opportunity for lab members or students new to protein expression and purification. Figure 3: The...express 10, 15, 20, 30, 40, 50, 60, 80 and 100 kD proteins in E. coli, either individually (~20,000 lanes...

...domain-containing proteins may be broader regulators of Cas13d activity. This property makes WYL proteins an intriguing... near a putative effector nuclease. The Cas13d proteins they identified have little sequence similarity...HEPN-independent. These enzymes also do not require a protospacer flanking sequence, so you can target virtually...advantages, including Cas13d-specific regulatory proteins and high targeting efficiency, both of which are.... Yan et al.: WYL-domain-containing accessory protein WYL1 increases RspCas13d and EsCas13d cleavage ...The majority of Type VI-D loci contain accessory proteins with WYL domains (named for the three conserved...Biotechnologies found that RspCas13d accessory protein RspWYL1 increases both targeted and collateral ...

...Fluorescent Protein-Based Tools Learn Some of the Pitfalls When Using Fluorescent Proteins Control Protein Activity...Fluorescent Protein Plasmids Find Fluorescent Protein Kits Find Empty Backbones with Fluorescent Protein Tags...researchers have used fluorescently tagged DNA-binding proteins to label certain loci, but this approach is not...telomeric sequences, Chen et al. successfully labeled protein-coding genes with both intronic and exonic gRNAs... ortholog was fused to a different fluorescent protein to create three distinct colors. The specificity...two hairpin sequences, which recruit fluorescent proteins BFP, GFP, and RFP fused to the respective hairpin...better. Adding an additional hairpin/fluorescent protein combination, for example, with a far-red FP, would...

...with BioRender.com.   Opto-G-protein coupled receptors  G-protein coupled receptors (GPCR) regulate... Sensing G Protein-Coupled Receptor Signaling. In D. Niopek (Ed.), Photoswitching Proteins (Vol. 2173,...tools: light-gated ion channels, optogenetic G-protein coupled receptors (OptoXRs), and photoswitches....are many different opsin (natural and engineered proteins responsive to light in combination with retinal...concentration gradient, and of activation inhibitory (ArCH) proton pump passing ions against the concentration gradient...similar concept, except the ligand is attached to a protein binding partner or nanobody that targets the native...

...suitable antibody against your target protein, consider fusing a protein tag to the target and using an epitope-based...Frozen Tissue-V2 v2. protocols.io. https://doi.org/10.17504/protocols.io.bs68nhhw Im, K., Mareninov, S...antibodies available at Addgene Protocols for Immunocytochemistry Protocols for other antibody applications...with tissues), where an antibody binds a target protein in your sample, then a fluorescently labeled secondary...then you use a microscope to see where the target protein is (localization) and how much there is (quantification...quantification). But what if you’ve got several interesting protein targets and you want to know if they co-localize...the nuclei. (See Brewer et al, 2022 for a sample protocol.)  Simultaneous IF Depending on your needs, your...

...extracellular proteins with community-based validation. They also have plans to expand to other protein technologies...foundation to begin experimenting. Protocols.io, JoVe, and Bio-protocol all work to provide more descriptive...Reagent Development - Steven C. Almo, Institute for Protein Innovation  1:18:45 - 2:01:18 (end) - Panel Alex...Researchers should be able to follow one another’s protocols and use one another’s reagents with the confidence...that aren’t always mentioned in static written protocols. It is therefore a bit ridiculous to expect that...to share in-depth information about reagents, protocols, analysis tools, and more. Being that this is ...Steve Almo’s talk about work at the Institute for Protein Innovation (IPI) (49:45 - 1:18:45), there are also...

...lines. Identifying less toxic envelope proteins or envelope proteins that target specific tissues are areas...that's gag, pol, and env), the isoforms of these proteins, as well as the viral long terminal repeats (LTRs... gene of interest. Consequently, the levels of protein expression tends to be much lower than those seen...including 293 were immortalized with adenoviral protein E1A, which has been shown to repress expression... 5'LTR, which requires the presence of the Tat protein to work, while 3rd-generation transfer plasmids... 5'LTR removes the requirement for the HIV Tat protein, thus decreasing the probability of creating replication-competent...envelope plasmid and a packaging plasmid encoding HIV proteins gag, pol, rev, and tat. 3rd generation systems...

...replaced the protein-based DNA-binding domain with a single-guide RNA sequence.  The Cas proteins, just like...catalytically impaired Cas9 protein to a DNA-editing enzyme to generate the prototypical base editor enzymes....CRISPR-Cas9 system. Nature Protocols 8:2281–2308 . https://doi.org/10.1038/nprot.2013.143 Seligman LM (2002...Maclyn McCarty’s experiment debunked the myth that protein is the carrier of genetic information and led to...enzymes, meganucleases, were discovered. These proteins are characterized by large DNA recognition sites...Era: Post CRISPR  CRISPR The strategy of using a protein to recognize DNA sequences is not ideal. For the...target new genomic sites. The discovery of the Cas proteins in 2005 was a major breakthrough in the world ...

...determine how Protein A and Protein B interact. You read extensive research on the two proteins and come up... an ideal antibody against Protein A and an ideal antibody against Protein B. The antibodies are both ...to detect Protein A and green fluorophore-tagged anti-mouse IgG2a secondary to detect Protein B and successfully... anti-Protein A antibody and an anti-mouse IgG2a secondary antibody to bind to the anti-Protein B antibody...are rat IgG1, then you could convert your anti-Protein B antibody to a mouse IgG2a Fc. The resulting chimeric...successfully visualize both proteins.     Figure 2: Two primary antibodies with the same isotype...

...can also be fused to a protein of interest to track the autophagy of that protein. Find the cytosol-targeted...mRFP-GFP-LC3 Tandem Fluorescent Protein, but following translation, this fusion protein is cleaved to generate ... Addgene.org Fluorescent protein plasmids and resources Fluorescent protein guide: Biosensors ...biosensor that’s comprised of two tandem fluorescent proteins: a relatively pH-insensitive RFP variant, DsRed.T3...autophagy biosensor. Its resistance to lysosomal proteases is key to its pH-dependent fluorescence. In neutral...variant at Addgene! mRFP-GFP-LC3 tandem fluorescent protein This autophagy biosensor contains mRFP and GFP ...to the autophagy-related microtubule-associated proteins 1A/1B light chain 3B, or LC3. LC3 is a commonly...

... expresses the endogenous protein, since folding of the recombinant protein may differ from the native... have since adapted it for a host of different proteins and small molecules from a variety of sample types...versatility lies in the user’s ability to modify the protocol to best fit their specific needs. Herein, we’ll...antibodies and thus are not suitable for low abundance proteins. Because the method uses a single antibody, it... use the same secondary antibody for any ELISA protocol that uses primary antibodies of the same isotype... and a series of wash steps removes non-target proteins and molecules from the assay. The user then adds...Advantages Disadvantages Direct Fast and simple protocolReduced chance of cross-reactivity Higher level of...

...GO works by affecting protein translation of different reporter proteins. Protein translation requires ...the translation of any detectable protein, not just fluorescent proteins. Figure 4: Reporters in Gene...codon to initiate protein expression, it can be used for an array of detectable proteins including fluorescent...monitors APOBEC (a family of cytidine deaminase proteins) and Cas9-mediated editing in real time (St. Martin... editing capability of the seven human APOBEC3 proteins, finding that APOBEC3A and APOBEC3bctd are the...reporter that is converted into BFP (blue fluorescent protein) upon CRISPR/Cas9 homology directed repair (HDR... a base-editing reporter. As the initiation of protein translation at the start codon ATG is universal...

...give rise to an adenine base editor. A massive protein engineering effort by Gaudelli et al. resulted ... restore antibiotic resistance. To find such a protein, they created a mutant library of E. coli tRNA ... ABE does not require any additional inhibitor protein like UGI in CBE. Base editors for the plant genome...based on the location of the ‘A’ relative to the protospacer adjacent motif (PAM). ABE7.10 can edit target...'A' if it lies in a small genomic window: at protospacer position 4-8 counting the ‘NGG’ PAM as 21-23 ...closer to the PAM, i.e., positioned at 8-10 in protospacer, ABE6.3 or ABE7.9 are better suited (Gaudelli...two Bsa1 sites to clone the single guide RNA (protospacer+scaffold) in a pol-III (OsU3) promoter driven...

...fluorophores to proteins, pioneers of this technique were able to directly detect protein/protein interactions... CFP-YFP was the first to be used to study protein-protein interactions and several other pairs have been..., Davidson MW. Fluorescent proteins for FRET microscopy: monitoring protein interactions in living cells...FRET-based biosensors, and interactions between DNA and proteins. FRET, in theory, is an easy technique, but it...to studying an unknown interaction between two proteins, these parameters are difficult to overcome but...used to create your choice of fluorescent fusion protein with your gene of interest: Plasmid Color Expression...About FRET: Piston DW, Kremers GJ. Fluorescent protein FRET: the good, the bad and the ugly. Trends Biochem...

...marks for safety.   Protein Delivery: Delivering reprogramming factors as protein is another way to avoid... vector-free Multiple transfections required Protein Delivery DNA-Free Low High Transgene-free and vector-free... inefficient Difficult to purify reprogramming proteins. Table 1: Key Features of Different Methods...are safer than integrating methods, with RNA and protein delivery considered the safest since there’s minimal...integration can occur. Okita et al describes a protocol for generating iPSCs by plasmid transfection where... following synthetic modifications are used to protect the RNAs from cellular defenses against ssRNA: ... exposing cells to exogenous genetic material. Proteins are delivered to cells by fusing them with peptides...

...Making proteins outside of a cell - cell free extracts Typically, if you want to make a protein – say ...DNA or transforming cells with green fluorescent protein, these activities are limited to those schools ...say green fluorescent protein – you first need the DNA sequence that encodes it. That DNA sequence then ...instructions it needs to produce that green fluorescent protein through transcription and translation. Unfortunately... live cells. The alternative is to express the protein in a cell-free manner. This is like taking an engine...systems contain all the parts needed to produce proteins without the need to handle a living, replicating...leaving them shelf-stable until water is added and protein production proceeds. FD-CF pellets are easy for...

...amino acids found in all proteins, so you can't distinguish the different proteins from each other. To distinguish...the surface or the inside of your cells. While proteins themselves can emit intrinsic fluorescence when...distinguish different target proteins in your flow analysis, you'll need to use fluorophores that attach...appropriate readout. A common method of labeling proteins is to fuse them with a fluorescent reporter, such...mCherry, through genetic modification. As your protein of interest will only occur in combination with... reporter's fluorescence. Another way to label proteins is through treating cells with fluorophore-conjugated...are completely different in nature (eGFP being a protein and FITC an organic molecule), their bleed is so...

...tool is the endoprotease Botulinum Neurotoxin B (BoNT/B), which cleaves the synaptic protein VAMP2, a protein...gas-filled protein structure that water-bound photosynthetic organisms use for buoyancy. These proteins allow...two fluorescent proteins and four repeats of a miRNA target site. One fluorescent protein (EBFP2) is expressed... protein essential for synaptic transmission. Cleavage of VAMP2 renders it inactive, thus inhibiting neurotransmitter... light-mediated? The researchers split the proteolytically active light chain of BoNT/B into two fragments...fragment with one part of an established pair of proteins (iLID and SspB) that undergo light-induced dimerization...function of microbes rely heavily on fluorescent protein genes. These optical reporters have limited deep-tissue...

...photoswitchable elements into many other types of proteins. Here, we’ll highlight a few recently-deposited...license. A naturally occurring microtubule-severing protein, Katanin is of course named for the samurai sword...monolayer of endothelial cells, showing cell protrusions and overlaps (arrows) after blue light illumination...optogenetic module to activate or recruit your protein of interest, maybe you want to inhibit it. Restricting...development and a knockout is not feasible or when the protein of interest supports a variety of processes in ...CIB1 clustering. Fuse either CRY2 or CIB1 to your protein of interest and express the other separately, then...generate CRY2-CIB1 oligomers that trap the fusion protein away from its usual location, preventing it from...

...Brainbow-1.0 construct contains three fluorescent proteins: RFP (red), YFP (yellow) and M-CFP (membrane tethered...Brainbow-1.1 scales up the system to four fluorescent proteins, with recombination events specified by three ...principles remain the same. The first fluorescent protein in the construct, OFP (orange), is expressed when...invertible segments, each with two fluorescent protein coding sequences in opposite orientations. Cre-...Cre is removed from the system, the fluorescent protein whose coding sequence is closest to the promoter...fluorophores used are fluorescently distinct, their protein sequences are highly homologous, preventing design... First, they screened a variety of fluorescent proteins to find those with ideal characteristics (low ...