...last year, Verkhusha added two far-red light photoactivatable (PA) near-infrared fluorescent proteins (FPs...fluorescent proteins, iRFP670 and iRFP720, as photoacoustic contrast agents for two-color imaging in animals...genetically-encoded probes by making them brighter, more photostable, smaller, and less toxic to cells. They’d like...751-4. doi:10.1038/nmeth.2521) Far-red light photoactivatable near-infrared fluorescent proteins engineered...: 10.1038/ncomms3153. PubMed) Multicontrast photoacoustic in vivo imaging using near-infrared fluorescent...

...This post was contributed by Richi Sakaguchi from Kyoto University, and Marcus N. Leiwe and Takeshi Imai.... For more details on this procedure, find the protocol here.   Tips and troubleshooting your Tetbow...thanks to our guest bloggers Richi Sakaguchi from Kyoto University, and Marcus N. Leiwe and Takeshi Imai...University. Richi Sakaguchi is a graduate student at Kyoto University. He is interested in developing genetic...High-resolution fluorescence imaging using SeeDB2." Bio-Protocol 8 (2018): e3046. Livet, Jean, et al. "Transgenic...

...Xin Zhou greatly expanded that list by using photoswitchable fluorescent proteins to create enzymes that...turned the enzymes back on. Xin termed these photoswitchable enzymes Fluorescent Light-inducible Proteins...various designs, Xin has been able to develop photoswitchable Intersectin, Cdc42, Raf1, Mek1, Mek2, Cdk5,... You can learn more about Xin’s work with photoswitchable components of the ERK signaling pathway in ...Resources on the Addgene Blog Learn More about Photosactivatable Fluorescent Proteins Use Light to Control ...

...sensor’s photophysical properties such as kinetics, ∆F/F0, SNR, dynamic range, photostability, quantum...signals upon sensing glutamate using both 1-photon and 2-photon imaging.   Figure 1: 1P and 2P excitation...membrane trafficking and localization. Improved photophysical properties The improved variants of iGluSnFR3...

...experiment check out Addgene’s protocol page, which has both protocols and videos for techniques in basic...about making your own competent cells and our protocols page to learn about bacterial transformation in...plasmid cloning Resources on Addgene.org Read our protocols for plasmid cloning Find our molecular biology...biology reference here Find molecular biology protocols here ...

...         Adipocytes [28] Keratinocytes [29] Photoreceptors [30] Otic hair cells [31] Neurons [32, 33... pluripotent stem cells from urine samples. Nat Protoc, 2012. 7(12): p. 2080-9. PubMed PMID: 23138349....from human ES and patient-specific iPS cells. Nat Protoc, 2015. 10(7): p. 941-58. PubMed PMID: 26042384....Generation, purification and transplantation of photoreceptors derived from human induced pluripotent stem...cerebral cortex neurons and neural networks. Nat Protoc, 2012. 7(10): p. 1836-46. PubMed PMID: 22976355...expression of insulin genes in liver and ameliorates streptozotocin-induced hyperglycemia. Nat Med, 2000. 6(5):...

...of Golden Gate cloning, the Haynes lab’s Protocols.io protocol describes an elegant system that allows ... Priode and Haynes, protocols.io. 2021. dx.doi.org/10.17504/protocols.io.brgcm3sw     New system developed...flanked by randomized nucleotides in place of a protospacer adjacent motif (PAM). They're used to determine...

... wavelengths of low photosynthetic activity. This still allows photosynthetically active wavelengths to... coral’s symbiotic dinoflagellates that are photosynthetic (Salih et al., 2000). Chromoproteins also enhance...Protein 101 blog series Learn more about FRET Find protocols and tips for making plasmid constructs Resources... (2000) Fluorescent pigments in corals are photoprotective. Nature 408:850–853 . https://doi.org/10.1038...

...included in commercial sets and most machines have a protocol that automatically performs the normalization ...normalization step.  Machines Most general-use protocols, unless otherwise specified, were likely developed on the...LightCycler, you’ll likely need to re-optimize protocols for that machine. (Teo, et al, 2002; see paper...an individually run sample when developing the protocol and optimized until the multiplexed and individual...samples or sample conditions change, the multiplexed protocol may have to be re-optimized.  While troublesome...eds.), Quantitative Real-Time PCR: Methods and Protocols, Methods in Molecular Biology, vol. 1160, DOI ...

...Using a spectrophotometer is among the faster methods of quantification. Many spectrophotometers have an... Antibody Purification Protocol Addgene's Video Antibody Purification Protocol Addgene's Antibody Guide...Antibodies in Basic Neuroscience Research. Curr Protoc Neurosci. 2020 Dec;94(1):e106. doi: 10.1002/cpns...

...able to come of age. The in vivo assembly cloning protocol In Vivo Assembly (IVA) cloning (Garcia-Nafria ... to be performed using a simple two-step, 2 hr protocol prior to transformation (Figure 1). As this approach...procedures are performed using the same three step protocol:  Fragment generation - Any linear DNA fragments...plasmid cloning Resources on Addgene.org Read our protocols for plasmid cloning Find our molecular biology...biology reference here Find molecular biology protocols here  ...

...IF, look for general IF protocols for your sample type. Starting with a protocol that usually works in ...details to help you plan and optimize your own protocols. Here we’ll start with just the first couple of...cause some confusion. Paraformaldehyde: Many IF protocols call for paraformaldehyde (PFA) as the fixative...datasheet to see if it has been tested in IF and if a protocol is available. When possible, try to start with...

...versatility lies in the user’s ability to modify the protocol to best fit their specific needs. Herein, we’ll... use the same secondary antibody for any ELISA protocol that uses primary antibodies of the same isotype...Advantages Disadvantages Direct Fast and simple protocolReduced chance of cross-reactivity Higher level of...sensitivityGood antibody availability Longer protocolLess specificityIncreased chance of cross-reactivity... High specificityHigh sensitivity Longer protocolIncreased chance of cross-reactivityChallenging to develop...

...genetic tools to the brain because of its low cytotoxicity, low immunogenicity, and the long-term expression...The issue with these FPs is that they are less photostable than GFP. In addition, antibody-based amplification...fluorescence. Furthermore, many of these FPs are cytotoxic, prone to aggregation and cross-react with anti-GFP... CA3 pyramidal neurons which are known to be notoriously difficult to label. It has also been shown that...structure and color variants." Cold Spring Harbor Protocols 2009.12 (2009): pdb-top63. PubMed PMID: 20150100...

...Enterobacter sp., Erwinia amylovora, E. carotovora subsp. Carotovora, E. herbicola, E. rubrifaciens, E. stewartii...salmonicida, Anabaena spp., Azospirillum braziliense, Azotobacter spp., Bordetella spp., Caulobacter spp., Enterobacteriaceae...Agrobacterium tumefaciens, Alcaligenes eutrophus, Azotobacter vinelandii, Brevibacterium methylicum, Caulobacter...Caulobacter crescentus, Desulfovibrio vulgaris, Erwinia carotovora, E. chrysanthemi, Escherichia coli, Gluconacetobacter...

...used in C. roseus, but processes specific to photosynthetic active tissues cannot be studied in hairy roots...leaf transformation: Check if there are working protocols available for your species This will save you ...first choice for establishing a new transformation protocol as it can easily show which tissue have been transformed...started. Once you have established a well working protocol in your lab, it can become a strong tool for your...assay by agroinfiltration of leaves." Arabidopsis Protocols. Humana Press, 2006. 225-229. PubMed PMID: 16739580...

...Platinum Gate TALEN Kit, please see the Yamamoto lab’s protocol for TALEN construction. Sakuma et al...plasmids, watch informational videos, download protocols, and more! Looking for background information ...phosphoprotein system, which includes detailed protocols provided by the Rinehart lab. Steinfeld et al...the most recently deposited tools, resources, protocols, and more. MoClo modular cloning system Synthetic...assembly and activity? The laboratory of Takashi Yamamoto has created a complete TALEN assembly system after...overlap, minimal protein aggregation, and high photostabilty. When multiple cassettes are integrated, each...

...at Harvard University currently studying the serotonergic system and behavior.     References 1. Southern...hybridization for detection of gene copy number changes." Protocols for Nucleic Acid Analysis by Nonradioactive Probes...): 205-226. PubMed PMID: 17332643. 4. Haruyama, Naoto, Andrew Cho, and Ashok B. Kulkarni. "Overview: engineering...engineering transgenic constructs and mice." Current Protocols in Cell Biology (2009): 19-10. PubMed PMID: 19283728...by PCR Resources on Addgene.org Check out Our Protocol Pages Find Molecular Biology Tools Analyze Your...

...Plasmid Cloning Protocols Find Virus Protocols Find Basic Molecular Biology Protocols Gibson Assembly...can be mixed yourself (e.g. see the Miller Lab Protocol). The Gibson assembly process can be used to assemble...commitments. These kits also come with specific protocols, suggestions for ratios of product to insert, ...

...proteins (NIR FPs) were derived from natural photoreceptors incorporating biliverdin IXα (BV) autocatalytically...from your source and to your camera.  Two-photon image of miRFP670nano3-expressing neurons in the... derived from bacterial phytochrome (BphPs) photoreceptors have higher apparent cellular brightness than...were derived from cyanobacteriochrome (CBCR) photoreceptors (Oliinyk et al., 2019). They were engineered...Molecular brightness vs. iRFP713, % Oligomeric state Photo-stabilityc, t1/2, s pKa Brightness in HeLa cells...