We narrowed to 883 results for: prot

...allow one to monitor compartment/protein dynamics or evaluate protein turnover rate. In many cases, however...Fluorescent Protein Plasmids and Resources Relevant Blog Posts Plasmids 101: Green Fluorescent Protein Which...studies or make a reporter construct. Fluorescent proteins (FPs) are also the foundation of multiple important...so thoroughly characterized, it turns out this protein has a few more secrets - during a collaboration...biological research. Photoconvertible fluorescent proteins Newly developed photoconvertible FPs like PsMOrange...repeat the process with these new photoconvertible proteins. With Sattarzadeh et al.’s discovery that GFP ...observed. Sattarzadeh et al. found that only the protonated form of GFP is photoconverted, so this technique...

...removes radio-labeled short-lived proteins primarily via proteasome degradation. Autophagy is then induced...supernatant (this is indicative of protein degradation). To control for protein degradation due to other pathways...an active form of the protein. To monitor autophagy, the Pho8Δ60 mutant protein is expressed. Pho8Δ60 ... Addgene.org Fluorescent Protein Plasmids and Resources Fluorescent Protein Guide: Biosensors Keima, ...studying autophagy Biochemical assays Long-lived protein degradation: This approach measures the degradation...degradation rate of radio-labeled long-lived proteins as a proxy for autophagy. Cells are cultured with isotope-labeled... to giving more fine-grained detail about what proteins or organelles are being degraded) and it is slow...

...molecule, the antibody binds to a protein, and then voila! Wherever your protein of interest is, a signal will...molecules to identify each protein of interest, this gel-based method identifies proteins solely by weight and... isn’t giving you enough signal to detect your protein of interest, it may be time to look at the indirect...antibodies. While a primary antibody binds to the protein of interest, a secondary antibody binds to the ...signaling molecules associated, indirectly, with each protein of interest.    Figure 1: The direct ... molecules are represented in light green; and proteins are represented in orange. Created with BioRender.com...three, secondary antibodies are indicated when the protein of interest is found in low amounts or if the conjugated...

... to test several protocols that have been described (protocol 1, protocol 2, protocol 3) to find the one...system.   See our protocol pages for full Lentivirus Production and Transduction Protocols   Once purified...Most of the protocols you will find will recommend using a polycation, like polybrene (protocol) or DEAE ... psi signal, the genes encoding the structural proteins could not be integrated into the new viral particles...plasmid), and 3) the vesicular stomatitis virus G protein (VSV-G) that improves the stability and broadens... one with the exception that the HIV accessory proteins that are not essential for the production of the... to be taken especially if you are working with proto-oncogenes or with human samples that have not been...

...particular the roles of proto-oncogenes, tumor suppressors, and their protein products. Proto-oncogenes consist...refers to 3 proteins (H-Ras, K-Ras, and N-Ras) encoded by a family of proto-oncogenes. Ras proteins are special...consist of a group of genes/proteins that, when expressed, normally encourage cell survival, growth and proliferation...dividing out of control. One may compare the roles of proto-oncogenes and tumor suppressors to the accelerator...respectively. In the case of a mutation that turns a proto-oncogene permanently “on”, (now an oncogene), this...and in the right places. Checks and balances: protecting against cancerous cells Luckily, there are several...several safety mechanisms in place to protect us from rogue cells when something does go awry. The process...

...labs already familiar with IHC/ICC protocols. A typical ExM protocol (Fig. 1) follows this structure: ...steadily growing in popularity with an abundance of protocols and examples of applications from across disciplines...several times its original size. Depending on the protocol, samples can be expanded to ~4-10x linearly! This... using more accessible imaging equipment, ExM protocols utilize relatively inexpensive reagents and are... Steps 2-4 are not found in standard IHC/ICC protocols and you might expect them to dramatically limit...the types of labels you could use. But many ExM protocols are compatible with conventional antibodies, chemical...chemical dyes, and genetically encoded fluorescent proteins (Gambarotto, et al. 2019; Tillberg, et al. 2016...

...codons that encode a protein may affect the rate of translation and thus protein expression. In fact studies...efficiency and thus protein expression without altering the sequence of the protein. As DNA synthesis has...expressing functional proteins in hosts that do not naturally express that gene. The protein you want to express...detrimental results to the protein. Codon optimization could affect protein conformation, folding and ...strategy for increasing protein production. If you are trying to express a protein in its native host there... Why is codon usage important? Translation and protein synthesis relies on transfer RNAs (tRNAs). tRNAs...cells. This was the first codon optimized membrane protein expressed in mammalian cells. In this study, codon...

...nontarget strand. Protein Cas: Different CRISPR systems involve different proteins, but many of them ...most famous of these proteins is Cas9 nuclease, but there are many other Cas proteins as well, both natural...recognized by the Cas9 protein from the appropriate species. Some Cas proteins, like LbCas12a or AsCas12a...you. We’ll cover the common terminology for the proteins, DNA, and RNA used in CRISPR.   Figure...tracrRNA (in some systems including Cas9), Cas proteins, and a CRISPR array. The CRISPR array is composed...spacer. In bacteria, these crRNAs are bound by Cas proteins (Cas9 shown here). The repeat sequence base pairs...natural and engineered. What these proteins all have in common is the ability to use an RNA guide to target...

...Rep proteins. Alternative splicing generates the different C-terminal regions of the Rep proteins. While... four Rep proteins share a common helicase and ATPase domains. To function, the Rep proteins form oligomers...viral capsid proteins VP1, VP2, and VP3, which together form the outer capsid shell that protects the viral... start sites generate the three VP proteins. The three VP proteins assemble into capsid at a 1:1:10 ratio...Assembly-Activating Protein (AAP) and the recently discovered Membrane Associated Accessory Protein (MAAP) from...the adenoviral helper plasmid which contains the proteins needed for the virus to replicate,  and the transfer...Replication (rep) gene The rep gene encodes four proteins that are required for viral genome replication...

...producing both a tagged protein from the tagged allele and an untagged protein from the untagged allele...of the protein gleaned from the literature and engagement with researchers who study the protein or structure...possible. This is especially true in the case of protein tagging to elucidate cellular structures. Overexpression...Overexpression methods flood the cell with protein, which can both interfere with a cell’s normal function... hard to visualize the precise location of the protein or structure of interest. Endogenous gene tagging...tagging and visualization of specific, individual proteins under endogenous regulatory control. But even ...transcribed and translated. For an N-terminally tagged protein, we used the same strategy, inserting the tag and...

...encodes a single envelope protein called rabies glycoprotein (G). G-protein-coated RABV infects cells ...order neurons, via entry mediated by the G protein/G-protein receptor. Once in first order neurons, the...recombinant rabies virus vectors. Nat Protoc. 2013;8:1583–1601. doi: 10.1038/nprot.2013.094. Pubmed PMID 23887178...line. By doing this, RABV expressing the viral protein EnvA on its surface will only infect Cre-positive...into these phenomenon has shown that the RABV G protein is necessary and sufficient for retrograde transmission...can be controlled by selectively supplying the G protein in trans only to starter cells (infected with G-deleted...cell. RABV replication will then occur and the G protein (supplied in trans) will be incorporated into the...

...GAG-POL polyprotein Nanoblades are formed when viral structural proteins, such as the MLV protein Gag, multimerize...the MLV Gag-Pol polyprotein, cleaves the proteolytic site connecting the two proteins, thus releasing ...purified by centrifugation. Gag-Cas9 fusion protein The MLV Gag protein fused to a Flag-tagged SpCas9 is incorporated...the unmodified MLV Gag protein. After Gag-Cas9 is incorporated, the MLV protease, supplied as part of the... envelope proteins To alter the cell tropism of Nanoblades, different viral envelope protein(s) are expressed...MLV)-like particles loaded with Cas9-gRNA ribonucleoproteins (RNPs). According to the authors, they named...with the the Cas9 portion of the Gag-Cas9 fusion protein. A single Nanoblade particle can be loaded with...

... are trying to remove from your sample is protein. Proteins absorb light to varying degrees across the...specific to proteins and why researchers traditionally use the A260/280 ratio as an indicator of protein contamination...DNA Quantification Protocol Recovering Plasmid DNA from Bacterial Culture Protocol    ...downstream experiments. The usual suspects, such as proteins, phenol, or guanidine salts, exhibit their own...how different contaminants impact these ratios.  Protein contaminants Whether you are purifying plasmid ... The A260/230 ratio can be a good indicator of protein contamination, but a number of other potential ...A260/280 ratio does not drop significantly until protein levels exceed 70% of the sample content (Loughrey...

...milk protocol is adapted from the 1979 Patterson Protocol. When published, the Patterson Protocol was ...causes the proteins and genomic DNA to precipitate. Plasmid DNA is free in solution. Proteins and genomic...plasmid purification protocol and guanidine isothiocyanate for the gel extraction protocol) disrupts the DNA...the same principles: cells are lysed, RNAs and proteins are removed to leave you with DNA. This DNA is...Product: plasmid DNA This kit-free plasmid miniprep protocol from Addgene follows a similar workflow as a column-based...you lyse the bacteria and denature the DNA and proteins in solution. The pH is then lowered using a renaturing...phenol chloroform extraction to remove remaining proteins and RNase. Finally, DNA is precipitated with alcohol...

...on designing entirely new proteins, improving our understanding of how protein folding works and creating... publish a human-designed protein that was not based on any existing protein structures, called Top7. ...novo designed proteins also helped researchers understand more about the biophysics of protein folding and....com. Top7 gets its name from “topology of protein” diagrams, which show the interactions between ...closer to achieving it. Once they designed the protein computationally, the researchers expressed, purified... a powerful proof of concept that this kind of protein design is possible. Although Top7 was designed ...created it have been applied to numerous other protein design goals, both from the Baker Lab and others...

... and a protein standard. The density of the antibody bands can be measured against the protein standard... Antibody Purification Protocol Addgene's Video Antibody Purification Protocol Addgene's Antibody Guide...predict the molar absorption coefficient of a protein. Protein Sci. 1995 Nov;4(11):2411-23. doi: 10.1002/...applications you are planning, cost, and the amount of protein you need to produce.  The most common yeast strain...pastoris) because it secretes fewer of its own proteins, making the purification process simpler. Due ...incubation time to maintain cell viability and increase protein production (Schwarz et. al., 2020). Harvest and...quantification. Many spectrophotometers have an IgG protein setting. If yours does not, be sure to set the ...

...decorated with a fluorescent protein remain functional. Attaching the fluorescent protein fusions allowed us to...produced by orchestrated movements of networks of proteins consuming molecular fuel sources. While the importance... can be generated by conformational changes in proteins? How can information propagate through a structured...rupture E. coli spheroplasts to release encapsulated proteins. Video by ACS. For a more cartoony explanation...addressed using R bodies: simple, force-generating protein machines that can extend and retract over many ...

...tagging proteins, as it can be fused to the N- or C-terminus (or even in the middle of proteins) and performs... including popular new approaches like PROTAC targeted protein degraders. But CRBN has been difficult ... produced by our partners at the Institute for Protein Innovation (IPI), part of a new collection of epitope...short, linear epitope derived from the P and V proteins of Simian virus 5 (SV5) (Randall et al., 1987)...drug ligands, and in ternary complexes with other proteins, establishing CRBNmidi as a promising tool for...future structure-guided drug design and studies of protein ubiquitination and degradation.   Find the CRBNmidi...for crystallographic and biophysical studies of protein degraders. Nat Commun., 15(1), 8885. https://doi.org...

...include proteins like nonstructural proteins 1 to 16, spike proteins, envelope small proteins, and membrane... Currently, plasmids containing all SARS-CoV-2 protein coding genes in E. coli and S. cerevisiae expression...mammalian expression vectors containing various spike protein variants are also available. Plasmids are continually...membrane proteins. Although all viral components are available in bacterial expression plasmids, glycoproteins...glycoproteins and other proteins that require post-translational modification may not be easily expressed ...plasmids The yeast expression constructs contain protein coding genes, tags, truncations, and codon optimizations...level expression and purification of SARS-CoV-2 proteins for downstream experiments and offer flexibility...

...specifically curated for protein expression in K. phaffii with special attention to protein modifications which...successfully for protein localization and transport studies in HEK cells, recombinant protein purification...great option for those interested in recombinant protein purification and metabolic engineering. If instead...library of parts but don’t need to add tags to your proteins, then the CIDAR Kit is an excellent option (Iverson...compatible with E. coli and/or cell-free systems. Protein expression in yeast MoClo is also for eukaryotes...eukaryotes! The technology has been used to optimize protein secretion and expression in the biotech workhorse...formerly known as Pichia pastoris). If your goal is protein isolation, then the MoClo Pichia Kit will get you...