...difluoride (PVDF) membrane, which is sturdier and retains the blotted protein better than a nitrocellulose...transfer, dry the membrane to maximize protein retention. Following this step, most PVDF membranes recommend...proceed with caution. Unlike PVDF, they can be returned directly to buffer without a re-wetting step. ...plan to reprobe! Just as your sample protein is retained better, antibodies will stick permanently to a...membrane once before any detection steps to avoid retaining residual antibodies. If you have already applied...membrane Alright, so you’ve dried your membrane and returned it to buffer, you’ve run and imaged your western...

... ShadowR, a novel FLIM-FRET acceptor Figure 4: ShadowR is used as a FRET acceptor to mRuby2 or mScarlet...microscopy-based Förster resonance energy transfer (FLIM-FRET) in living cells (Figure 4). They created ShadowR...inhibit their activity. ShadowR can be used as a FRET acceptor in combination with mRuby2 or mScarlet ...Fluorescent Protein 101 blog series Learn more about FRET Find protocols and tips for making plasmid constructs...our Molecular Biology Reference Find plasmids for FRET References Blanquet RS, Phelan MA (1987) An unusual...

...generated a series of retroviral vectors expressing tagged dCas9 and guide RNA. The retroviral enChIP system ...necessary to isolate specific genomic regions while retaining molecular interactions. For this purpose, we developed...requested plasmids for enChIP through Addgene. Retroviral Expression System for enChIP using CRISPR Addgene...Addgene: Your lab recently developed a retroviral enChIP system using CRISPR technology. Can you describe...Associated with an IFNgamma-Responsive Promoter by a Retroviral Expression System for enChIP Using CRISPR. Fujita...

...based on our most accurate interpretation of the world, and this interpretation should not effected by claims... government and, consequently, politics. More concretely, scientists gather information about reality ...however, inconvenient data must not be ignored. Returning to an earlier analogy, no amount of shouting will... of depending on scientists or politicians to interpret it for them. Other popular reasons to march include... not in the job of having evidence’ on Trump’s wiretap claim The Chronicle of Higher Education: “March...

...positive and negative controls to ensure proper interpretation of results. A positive control is generally.... This information is critical for correctly interpreting experimental results.  Replicates Replicates... needs to be addressed. In order to correctly interpret your results, the signal from your target needs...autofluorescence, also interferes with data interpretation and can be addressed with a label control. ...autofluorescence and should be considered in the interpretation of results. Include the label control for all...membrane. A loading control is critical for data interpretation, especially when changes in protein expression...

...include the quality control of secretory proteins in the Endoplasmic Reticulum, optimization of fluorescent...committee members provide expertise in designing and interpreting experiments, as well as career guidance. In ... controls? Will these controls enable you to interpret your results? Will the experiment actually answer...mentor, manage, communicate, and teach The dirty secret about becoming a group leader is that people are...student. Learn to teach how to design, perform, and interpret experiments. You'll be better prepared for your...

...plasmid deposit please sign up here.        A retroviral mouse CRISPR knockout library Article contributed...issue, Sarah Teichmann and colleagues created a retroviral CRISPR knockout library using gRNAs designed ...designed by the Kosuke Yusa Lab. This retroviral library was used to transduce T helper type 2 (Th2) cells from...differentiation factors as the screen readout. This retroviral mouse CRISPR pooled library can be used for screening...These include Arch(D95N) and QuasAr, as well as the eFRET sensors, QuasAr-mOrange and MacQ-mCitrine. All of...

...energy transfer (FRET)-based executioner caspase reporters. In these reporters, the FRET pair is linked ...emission. FRET and fluorescence based reporters however have several limitations. First, FRET signals are...a clear pattern in the developing forebrain and retina. This pattern was consistent with TUNEL staining...

...microvesicles (approx. 40 - 150 nm diameter) that are secreted from the cellular membrane into the culture medium...demonstrated in different cellular environments such as retina or glioblastoma). They also found that exo-AAVs...AAVs (Schiller et al., 2018). Exosomal vesicles secreted from producer cells also contain cellular macromolecules...deafness (György et al., 2017), exo-AAV2 in mouse retina after intravitreal injection to transduce bipolar...vector mediates robust gene delivery into the murine retina upon intravitreal injection." Scientific reports...

...information, accession numbers, sequences and lineages, FRET efficiencies, additional references, etc...) and...frequently) for the sake of comparison or ease of retrieval.  These collection pages can be kept private, ...be downloaded in JSON or CSV format to quickly retrieve all of the information in the database relating... applications, a REST API allows programmatic retrieval of the majority of information in the database...all fluorescent protein plasmids Find plasmids for FRET Check out the Michael Davidson Fluorescent Protein...

...GBC) 85968 Barcode Human Weissman 3rd N/A N/A Retroviral Mouse Genome-wide CRISPR Knockout Library 104861...104861 Knockout Mouse Teichmann N/A (retroviral) 5 90,230 RNA-Binding Protein Pooled CRISPR Knockout Library...

...“no.” When there was room for misinterpretation, that misinterpretation was definitely happening. Our ...were leaving these meetings with different interpretations of what we discussed and decided. I needed ...Whether or not the container was barcoded (this is pretty Addgene specific) The types or purposes of each...that seemed simple, but they could in fact be interpreted in several different ways depending on the team...

..., the solution is diluted, complicating the interpretation. Because of this, these methods only give a...direct the fluorescent protein to the Endoplasmic Reticulum (ER). If homodimerization occurs (most likely ...structures known as Organized Smooth Endoplasmic Reticulum (OSER) whorls are visible. These whorls can be...CytERM-mTurquoise2, and CytERM-dTomato. When using and interpreting the results of the OSER assay, you should keep...Amsterdam) where she develops genetically encoded FRET based biosensors.     Additional Resources on the...

...classification. Lentivirus (a type of retrovirus) and gamma retrovirus require more caution as they are integrating...components of a virus into different plasmids. For a retroviral system this looks like the genetic cargo in one... researcher.   Figure 1: Separation of retroviral packaging components onto distinct plasmids for...common types of viruses grown in the lab are gamma retrovirus, lentivirus, adenovirus, and adeno-associated...

...MSCV-IRES-EGFP IRES Retroviral pMSCV-pBabeMCS-IRES-RFP IRES Retroviral pMSCV-IRES-YFP II IRES Retroviral pCMMP-MCS-IRES-Puro...pCMMP-MCS-IRES-Puro IRES Retroviral pEF1a-IRES-Neo IRES Mammalian MSCV-IRES-Luciferase IRES Retroviral pWPI IRES...

...life science literature is a litany of false interpretation in cell culture for this reason. Unreproducible...just a few publications which have since been retracted due to cell line contamination: Epithelial Mesenchymal...even decades after it is initially isolated. Regrettably this is not usually the case if you obtain a ...colleague. Time and money are wasted, papers are retracted and there is a loss of reputation that can impede...working vials have been used up. This ensures you can retain the maximum stocks and use of the original Cell...

...using HR was found to block the acquisition and retrieval of contextual fear memories [5].  In a modified...other regions of the brain may be involved in retrieval of remote contextual memories. This study is an...real-time inhibition), and behavioral (remote memory retrieval) specificity. Optogenetics can be used to more...Twitter @derekpsimonphd.    References 1. Stuber, Garret D., et al. "Excitatory transmission from the amygdala...2012): 159-172.5. Goshen I, et al. Dynamics of retrieval strategies for remote memories. Cell. 2011;147...

... constant variables that enable the correct interpretation of the effect of the independent variable you...controlled for in order to eliminate any alternative interpretation of the results. Typically, plasmid-based experiments...non-targeting control is critical to the correct interpretation of the results because it provides an important...limitations and you should be cognizant of these when interpreting your results. The key to biological controls...multiple replicates, and ultimately getting interpretable results: Figure 4: Expression Level of Gene ...

...determined by the organized smooth endoplasmic reticulum (OSER) assay. A major advantage of mCyRFP1 is ...ability to be coexcited along with EGFP while retaining an emission spectrum that is easily separable ...makes mCyRFP1 a powerful tool for use as a donor in FRET (fluorescence resonance energy transfer) and FLIM...Lin and Yasuda groups use both mCyFP1 and EGFP as FRET donors in experiments simultaneously measuring the...mammalian mCyRFP1 or CyRFP1 expression plasmids, FRET constructs, and CaMKII and RhoA sensors. Laviv,... of ~50% and confirmed that the fusion protein retained the PAM properties of the individual Cas9. Kim...

... Photoreceptor Cells AAV2, AAV5, AAV8 RPE (Retinal Pigment Epithelium) AAV1, AAV2, AAV4, AAV5, AAV8...AAVs may move toward the cell bodies of neurons (retrograde transport) or towards the synapse (anterograde...serotypes 1, 8 and 9, not only do they undergo retrograde and anterograde transport, but they also have... specific cell populations. Find Ready-to-Use Retrograde AAV Preps at Addgene 2) Comparison of genetically...share conserved mechanisms for anterograde and retrograde axonal transport." Human gene therapy 25.8 (2014...