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... of interest is not available in a pure form, you can perform relative quantification, which measures ...If your protein of interest is available in pure form, you can use it to create a standard curve on your...different than the validation assays, we recommend performing your own validation assays. If you’re using previously...chemical-based. Before selecting one, check your antibody information. Many antibodies will have a recommended blocking...temperature for 1–2 years and are available in formulas specific to your choice of detection methods. ...Aldridge et al., 2008).  Woah! That was a lot of information to cover. But don’t worry — the next time you...

...’t be lazy about splitting cells, instead try to form a routine. Twice a week often works for most fast-growing...

...transsynaptically). This is because the recombinant forms of these viruses typically used in research aren...after, we learned about organs and the jobs they perform. This became foundational later on when we studied...

...subtilis genome, the Mobile-CRISPRi ICE element forms a conjugative plasmid-like DNA circle and is transferred...

...case, the larva will metamorphose into its adult form. In all, it takes roughly 10 days for an egg to ...Some resources will direct you to the latest information about gene function, while others offer fly stocks...Flybase.org is absolutely essential. You can look up information about your fly gene using the search box in the... started. Any given gene’s webpage will have information on where the gene is expressed within the fly...website as well. Additionally, they have useful information on topics such as choosing the right balancer...

...working day, including appropriate MTAs and customs forms. Of course, ordering the plasmids is the easy part...

...ligase complex. Finally, when auxin is added in the form of auxin hormone indole-3-acetic acid (IAA), the...behave with a new protein, but you can make an informed choice!   References and Resources References...

...exogenously to measure a phenotypic response in the form of fluorescence or voltage indicators.   Viral... here!). The only issue with these GEVIs? They perform poorly in two-photon conditions, which are necessary...

...resides in and potentially which other proteins it forms complexes with. These “colocalization” studies give...

...transcriptional start site of a gene of interest, Cas9 forms a steric block that prevents RNA polymerase from...

...MMEJ (microhomology-mediated end-joining)? MMEJ, a form of alternative end-joining, requires only very small...’s become clear that the power of this editing platform lies in the diversity of nucleases and their applications...

...the two halves are in close proximity, they will form the full FP and fluoresce. Split FPs can be used...

...fill out our form to let us know what you would like to see available in viral prep format. Please note... protein, the virus can replicate but the newly formed viruses are unable to infect any neighboring cells... AAV can be used with modified rabies virus to perform retrograde neuronal tracing. Note : Addgene does...

...antibody fragments in the form of scFvs A simple vector system to improve performance and utilisation of recombinant...added to this list? Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve this...Find information on Addgene's collection of plasmid-based antibodies, including monoclonal antibodies... Synthetic single domain antibodies for the conformational trapping of membrane proteins. Elife. 2018...

...are required for proper packaging of AAV, but also form secondary structures that are prone to deletion ...and XmaI restriction sites). Another way is to transform AAV transfer plasmids into bacterial strains, ...found in the culture media, you should consider performing PEG precipitation. Instead of sonicating the ...

...choice, you should perform a functional assay to make sure your protein isn’t forming inclusion bodies ...

...antibody-target protein immune complexes are allowed to form. Following incubation, beads are used to immobilize...-40 or triton-X, to preserve native protein conformations and interactions. For difficult to release proteins... in the lysis buffer, keep samples on ice, and perform the IP at 4oC. When choosing the capture antibody... recognizes the target protein in its native conformation. Frequently, the antibodies used for a standard... are completely removed. Wash steps should be performed even with a pre-cleared sample.  Eluting the target...and negative controls for an IP. When possible, perform the IP in parallel with both a sample known to ...order to identify any non-specific interactions, perform parallel IPs with beads only and an isotype control...

...protein interaction partners (PIPs), the FP fragments form the complete FP because the PIPs they’re tethered...to bring them into close proximity (Figure 1). Formation of the complete FP results in a fluorescent signal...selection of FP fragments and, preferably, some information about the structures of the tested PIPs. Many...fuse the FP fragments. In the absence of this information, you can fuse each FP fragment to the N- and ...your results. Unfortunately, without structural information, constructing appropriate controls may requires...

...synthetic oscillator, bistable network, stripe pattern-forming incoherent feed-forward loop. Find the plasmids...systems at Addgene! Schematic of the evolution platform to generate TEV protease variants. Catalysis by...the right places, which can provide misleading information on where exactly a structure or protein is located...dissolved at a higher concentration, and results in uniform NB biotinylation. Once induced, the NBs could be...developed fluorescently labeled piggyBac vectors for uniform and sustained expression of single or multiplexed...

...chromatin, the complex of DNA and proteins that forms chromosomes. These long range DNA interactions are... are often detected with chromosome conformation capture (3C) based methods, such as 5C or Hi-C (Han et...generated with current 3C methods provide useful information about chromatin interactions, they are grainy...for answering more questions about chromatin conformations. The Xu lab presented three ways to use CAPTURE... capture of chromatin interactions The Xu lab performed a proof-of-concept experiment to show that CAPTURE...original CAPTURE method. Both CAPTURE methods outperformed other 3D chromatin methods such as ChIA-PET ...is detection of temporal changes in chromatin conformation, like those that occur during development and...