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...to the right indicates a higher signal, or upregulation, and a peak to the left indicates a lower signal...anything to the right of it, marking it as GFP+. Quantification for such graphs could simply be the percentage...case for you, switching to a contour plot should indicate where the two populations diverge. The smear ...place your gate based on the dot plot would be complicated.  Figure 2: (A) a smeary dot plot showing...Figure 4 compared to the dot plot in Figure 2. Quantification using MFI makes it easier to identify and study...Research Assistant in Arlene Sharpe’s lab at Harvard Medical School.   More resources on the Addgene blog Antibodies...addgene.org Addgene's Antibody Guide rAb Affinity Purification Protocol Video rAbs in the Addgene repository...

...systems are available at Addgene! Chemically inducible promoters Chemically regulated promoters are among ...sensitive expression systems are typically less leaky than chemically induced promoters; they show near-zero...transcription is turned ON. Types of inducible promoters Chemical agents, temperature, and light are all examples...prokaryotic promoter often used for bacterial protein purification. When arabinose is absent, regulatory protein...decreasing promoter leakiness.   Other examples of chemically induced promoters include positive inducible ... promoters only work in prokaryotic cells and typically only in the same or closely related species from...

...cost-prohibitive for many labs, and are often impractical for small projects. Most researchers instead ...involves splitting up a sample into thousands of physically isolated partitions (such as droplets or wells...Since there are many thousands of partitions, statistical modelling can be used to determine the actual..., does not require standard curves, and can theoretically detect a single target molecule in a sample....a gene of interest is one of the most common applications of precision genome editing, the remainder of...single molecule resolution of digital PCR allows classification of individual target molecules based on their...single wells of a 96-well plate without prior quantification. Comparatively, mismatch assays require between...

...red fluorescent proteins) allow simultaneous identification of more than one protein of interest. However... species (bottom panel in Fig. 1). Our results indicate that the tag protein may have an important influence...plasma membrane. Note that the use of the HA tag significantly improves the transport of hM4Di to the plasma...However, in some cases the fluorescent signal may be critical for the experiment. Since the fused protein could...fluorescence expressed in the neuropil conceals identification of opsin-expressing cells). For other things...Research Center. Using a combination of in vivo and anatomical techniques, her research aims to gain a deeper...Rodgers L, Wilson IA, Lerner RA, Wigler M (1988) Purification of a RAS-responsive adenylyl cyclase complex...

...and success. Customer and technical support Our customer and technical service brands continue to ...to keep up with the demand with excellence and dedication. We don’t have worry or wonder if providing...projects allows us to support more new projects. Diversification is happening. The Addgene team will have some... We score ~99.9% helpful and the customer and technical service teams put 100% effort into helping our...we might do. I am grateful every day for their dedication, flexibility, innovative thinking, and openness...Addgenies support a high collaboration, high communication, high efficiency, high quality, low meeting,...about the Addgenies. I never worry about their dedication, excellent intentions, or incredible drive to...

... microscopy. This application of AHA labeling is known as fluorescence noncanonical amino acid tagging...PhD students at the Max Planck Institute for Biophysical Chemistry, Göttingen in Germany. I attended this... symposium seeking inspiration to resolve the technical hurtle of monitoring protein turnover of specific...technique applies metabolic labeling using a noncanonical amino acid, azidohomoalanine (AHA). In the technique...DNA strands take part in rolling circle DNA amplification. The amplified DNA segment is labeled with fluorescent...directed toward AHA. Now the rolling circle amplification could only take place if the two antibodies ...interested in neurobiology.  She is passionate about communicating science to all audiences. Right now, she is ...

...repurpose popular GFP reporter lines for more sophisticated experimental manipulations, saving the time ..., 98% of TdTomato expressing cells were GFP+, indicating a robust, but specific system. T-DDOGs also successfully... types of proteins. They started with another biological workhorse, Cre recombinase, which induces recombination...systems are easily adaptable to neuroscience applications, including optogenetics, and they should make...Tang note that their scaffold system is likely applicable to many types of proteins. For proteins with ...Could Cas9-DOG be developed soon to allow genome modification only in a small subset of cells? Other proteins... PMCID: PMC4096992. Find plasmids from this publication at Addgene. 2. Tang, Jonathan C.Y., et al. “...

... work around this is to "domesticate" your fragment: PCR-based amplification can be used to create silent...and exciting applications, this system is an incredibly powerful tool for cloning complicated constructs...plasmid (Option 1) or added using PCR-based amplification (Option 2). Advantages of Golden Gate cloning...restriction sites that may not be amenable to "domestication", you might want to consider using an alternative...design of flanking overhangs. Although there are theoretically 256 distinct flanking sequences, sequences that...high-efficiency step. References: Sequential amplification of cloned DNA as tandem multimers using class-IIS...

...generated in vitro using synthetic genes that are typically expressed from a plasmid or from an integrated...cells, for antibody production and subsequent purification.  Academic laboratories and companies have already...share, making recombinant antibodies the most practical choice. Hybridomas are much tougher to ship, often... often lack sequence data, and may genetically drift over time. Antibodies made from animals are limited...the ease with which scientists can now improve genetically-encoded research tools, it is shocking that the...Begley CG, Ellis LM (2012) Raise standards for preclinical cancer research. Nature 483:531–533 . https:/...Edition. 2104. Cold Spring Harbor Laboratory Publications, New York. Helma J, Cardoso MC, Muyldermans ...

...very useful in the event that you will not be physically present at your poster at all times, so other... sections should be as concise as possible, communicating only the most important results to your audience...have the advantage of being able to verbally communicate your work, so you don’t need every little detail...readers can navigate your poster and follow the logical progression of your work, and you can accomplish...offers free poster templates, although most are vertical.  Large central box. Another commonly used and...presentation. Portrait. This style displays a poster vertically rather than horizontally. Instead of displaying.... Shapes. In this type of poster, written and graphical information are inserted into larger circles or...