... be run in duplicate or triplicate. The closeness of the data points generated by replicate values provides... the standard dilution series in duplicate or triplicate. Replicates allows you to calculate the curve...blot. Standard curves are critical for a number of popular scientific applications, and the quality of a ... deviation of each replicate measurement divided by the mean of all the replicates multiplied by 100. ...regularly, and mix samples very well.  Use duplicates or triplicates to increase the accuracy of your experiment... value. Though it varies from assay to assay, typically the experimental value should be within 15% of...correlation between the actual values and the predicate values and measures how well the data points fit...

...caudal NST. (A-C) Low-magnification images showing viral spread. (D-F)  High-magnification images immuno-stained...vector performance. Yet, most of the technical data needed to replicate and build on others’ results is available...Produce Chemically and Functionally Distinct Vectors. Molecular Therapy - Methods and Clinical Development...and community concerns AAV vectors are complex biological reagents; many factors, like storage buffers ... right AAV for your system or to successfully replicate published works. Of particular importance to these...improve our Addgene Data Hub, 76% of respondents indicated they wanted to see more negative data (Figure ...out of fear it points to flaws or gaps in their technical ability. But, especially for AAV data, that may...

...and often cannot eradicate metastasis or eliminate the probability of a relapse. Chemical and radiation ...the tumor, this results in frequent physiological and morphological changes. Tumors differ in their genotypic...-level barricades put up by cancer. As mentioned in this article, cancers differ in practically every ...immune response to the development of tumors is a complicated and dynamic phenomenon that is interestingly ...either due to age, infections, or other strenuous biological changes. In contrast to the acute immune responses...themselves from attack. It is a region of incessant chemical activity, and consists of other cell types, especially...attack the cancer cells are also suppressed by chemical signals such as IL-8, IFNγ, and colony stimulating...

..., can be expressed from lentiCas9-Blast. Purpose of LentiCas9-blast LentiCas9-blast is a Cas9 expression...single-vector systems typically express Cas9 and a gRNA from the same plasmid. LentiCas9-blast virus can be...began with only a few inventory items offered domestically, but by the end of 2016, we expanded our viral...2016 were (drumroll please)... Top Lentivirus: lentiCas9-Blast Most of Addgene’s lentiviral inventory is... preps. Cas9 is the headliner of most CRISPR applications and can be expressed from your cell line of ...2014). It is a dual-vector system, meaning it is typically used in conjunction with a second vector that ...several benefits. First, the low viral titers typically associated with Cas9-expressing lentiviruses will...

...By Alyssa Neuhaus Genetic modification of microglial immune cells is critical for central nervous system...mice. B) Quantifications showing high microglia specificity of GFP expression for the indicated ssAAV5s...ssAAV5s. C) Quantifications showing high microglia transduction efficiency for the indicated ssAAV5s. Figure...Emily Bentley Building on their CROPseq method for optical pooled CRISPR screening, Paul Blainey’s lab has...compatible with enrichment, single-cell sequencing, and optical pooled screens in human cells. Addgene offers vectors...3’ long terminal repeats (LTR). The 3’ LTR is duplicated during lentiviral integration, producing a second...described in Anzalone et al., 2019, including modifications to the Cas9 or reverse transcriptase enzymes...

... University where she engineers genetically encoded voltage indicators. Tian developed a video-based high-throughput...red‑shifted archaerhodopsin‑derived genetically encoded voltage indicator which had improved performance in...the improved voltage indicator coming from the screen to understand mouse cortical and hippocampal neural... Haoyi Qiu He Tian Every two years, we seek applications for awards to early career scientists using ...virtual conference attendance. We also opened up applications to research technicians and undergraduate researchers...the brain. “We hope that iGluSnFR3 will enable significant steps forward in our understanding of synaptic...the chromophore that are susceptible to redox modification, the MPST hydrosulfide can be transferred to...

...in PubMed and choose the appropriate publication. This publication will be listed at the top of the final... "Published plasmids" or "Unpublished or pre-publication plasmids" and set a desired distribution status...quickly associate your plasmids with the original publication where you described them. Search for your article...plasmids. You can submit "Unpublished or pre-publication plasmids": Unpublished plasmids or plasmids that...accepted. Addgene also encourages submitting pre-publication plasmids so they can be available when the paper...preprint submission, we are happy to update the publication information once the manuscript is published ...your desired distribution status: Hold for publication: we will initiate our MTA process and perform...

...likely that the shifts caused to the practical and philosophical landscape of the scientific enterprise...Director of Media Communications, Harvard University Ray Howell -Founder of Howell Communications Philip Brenner.... Revitalizing biomedical research: recommendations from the Future of Research Chicago Symposium F1000Research...and keynote by former congressman and CEO of the American Association for the Advancement of Science, Rush...solutions. The event also culminated in a joint American Society for Biochemistry and Molecular Biology...union dues. Time will tell the extent of the implications of this ruling and whether graduate students...focus toward advocacy and action with sessions specifically chosen to better define productive routes for...

...Mechanisms of epigenetic modifications The main mechanism of epigenetic modification is chromatin remodeling...in this study,epigenetic changes were statistically significantly associated with reduced life-span, but...methylation in living cells." Journal of the American Chemical Society 126.19 (2004): 5982-5983. PubMed PMID...literally means ‘on top of the genome’). Epigenetic modifications can result in phenotypic changes without any...any change in genotype. Epigenetic modifications play a large role in turning genes ‘off’ and ‘on’, signaling...becomes. This prevents the cellular machinery from physically interacting with the genes in the compacted region...two main mechanisms, either through histone modifications, which can influence the transcription of genes...

...arrows indicate a conversion directly catalyzed by the base editor. Dotted red arrows indicate the change... point modifications. CRISPR base editing persists through multiple cell divisions, indicating that this...target modification in primary T cells, making them a promising tool for cell therapy applications. Dual...Base editing publication highlights In the table below, you'll find a list of publications we've highlighted...Early CRISPR applications were often limited by the low editing efficiency of homology-directed repair...all. Base editing to the rescue! Base editors chemically modify DNA bases without breaking the DNA backbone...following base editing and DNA repair. Black arrows indicate natural cellular processes. BER = base excision...

...which provides the AAV replication (rep) and capsid (cap) genes. AAV replication uses the host’s polymerase...this AAV purification video! It didn’t exist when I did my first iodixanol gradient purification, but I ...margins of my protocol. While these tips weren’t critical to the experiment, they definitely made my life...In this post, I’ll share some AAV production, purification, and titration tips, while also summarizing ...adenovirus genes which help mediate AAV viral replication; and 3) the transfer plasmid which encodes a ... that aids in the homologous recombination of duplicated regions like the ITRs. Restriction digest screening...consider performing PEG precipitation. Instead of sonicating the cell pellet, I lyse the cells with this AAV...

...blogger Jessica Polka, a Postdoctoral Research Fellow with Pamela Silver.  Most types of biological motion...these applications, purified R bodies could also perform as actuators in microelectromechanical systems... with new ways to interact mechanically with their environment.   Jessica Polka is a Postdoctoral Research...Systems Biology at Harvard Medical School. Follow her on Twitter @jessicapolka.     References 1. Kanabrocki...it to suit our purposes? In search of simple biological machines R bodies can repeatedly extend and...R-bodies, check out this video. Most types of biological motion (whether endocytosis, vesicle trafficking...to pH changes without the need for ATP or other chemical fuel sources. Like an actin filament or a microtubule...

...3’ end binds template DNA for PCR amplification, while modifications and homologous sequences are encoded...multiple modifications A key feature of IVA cloning is the ability to combine multiple modifications. Any ...’ Check your amplification by gel electrophoresis. In our experience, good amplification after PCR means...gain traction? It is likely that both vector amplification with low fidelity polymerases and the cost of...this approach requires no special reagents or purification kits, it is low cost and can be immediately ...to produce the linear fragments, as desired modifications and homologous regions can be encoded in the...parental DNA (37 °C for 15 mins). This enzyme specifically cuts methylated DNA, so it will selectively ...

... ensure they bind their antigens specifically. Possible applications for validation include western blot...addgene.org Addgene's Antibody Purification Protocol Addgene's Video Antibody Purification Protocol Addgene's Antibody...cells can perform human-like post-translational modifications that bacterial and most yeast cells cannot. ...production system may depend on what downstream applications you are planning, cost, and the amount of protein...secretes fewer of its own proteins, making the purification process simpler. Due to its high yields, E. ...using a mammalian production system, cell lines typically used include Human Embryonic Kidney (HEK) cells...production (Schwarz et. al., 2020). Harvest and purification At its core, harvesting recombinant antibodies...

...next-gen sequencing. ChIP is an incredibly useful application for exploring these interactions, and can give... protein interactions, including epigenetic modifications. But it's not an approach for the faint of heart...your sample before moving on to the next step. Typically, this can be done by cutting the tissue sample...into random, small pieces of roughly equal size, typically between 200-1000 bp. This is to ensure that your...background information. Your options for this step are sonication, which uses high-frequency sound waves to break...break up the DNA, or enzymatic digestion. Sonication provides a truly random digestion, but is harsher ...to the selective nature of the cutting. While sonication is usually the preferred method, enzymatic digestion...

...2024 American Chemical Society. Both PILATeS (18 kDa) and STReTCh (11 kDa) are significantly smaller...cofactors or using significantly truncated forms that limit its functionality. To make biophysical and structural...Sensors for Bioluminescent Readout of Mechanical Forces in Biological Systems. ACS Sensors, 9(7), 3489–3495...disruption in M. abscessus  Bioluminescent reporter of mechanical forces   Epitope tag antibody collection grows...performs well in many different antibody-based applications.  IPI’s recombinant version is a fusion of mouse...properties, and is amenable to a wide range of biophysical assays (Kroupova et al., 2024). They were able... Cereblon construct for crystallographic and biophysical studies of protein degraders. Nat Commun., 15...

...eukaryotic secretory pathway."Biochemical and biophysical research communications 430.3 (2013): 1114-1119. ...referring to the evil side of a mystical "Force," this quote is equally applicable to many microscopy experiments...within these compartments undergo significant post-translational modifications including glycosylation, disulfide... the cytoplasm? Or more troubling, what if a significant fraction of the fusion protein molecules fail...time. Previously, we have described a number of practical considerations that should be taken into account...human (and most eukaryotic) proteins localize to chemically distinct subcellular environments, including ...are equally susceptible to these non-native modifications, which can all potentially affect function [...

...fluorescent genetically encoded voltage indicator Article contributed by Shreya Vedantam Genetically encoded... for its biotechnological applications including conversion of CO2 into water and electrical energy generation...can be reconstituted in vivo to form a protein identical in sequence to the unmodified enzyme. Each half...tested the base editors in vivo and achieved therapeutically relevant editing efficiencies in the mouse ... base editors at Addgene. Levy et al., Nature Biomedical Engineering, 2020. PubMed PMID: 31937940. Detection...encoded voltage indicators (GEVIs) are proteins that detect and output the voltage changes in neural activity...fluorescent optics. Existing red fluorescent voltage indicators, however, fall short compared to GFP-based sensors...

...HIV. A notable landmark study showed significant clinical efficacy against a form of blindness known as...patients with clinical diagnosis of LCA and mutations in RPE65 (n=3), showed significant improvements ...and other ongoing clinical trials support the utility of AAV as a broadly applicable gene delivery tool...Almost all patients have achieved persistent immunological reconstitution with a normally functioning T...translating gene therapies from conceptual design to clinical trials involve identifying a therapeutic gene,...carcinogenic profiles have limited their usability in clinical settings. On the other hand, vectors derived from...Unlike most naturally occurring viruses used in clinical trials, wild type AAV is not associated with any...

...calcium indicator (Shemetov et al., 2021). In addition to FRET, intensiometric calcium indicators NIR-GECOs...characteristics, performance, and applications of near-infrared calcium indicators in this recent review (Shcherbakova...expertise is development and applications of fluorescent proteins, genetically encoded biosensors, and optogenetic...Qian, Y. et al. A genetically encoded near-infrared fluorescent calcium ion indicator. Nat Methods 16, ...Shemetov, A.A. et al. A near-infrared genetically encoded calcium indicator for in vivo imaging. Nat Biotechnol...Shcherbakova, D.M. Near-infrared and far-red genetically encoded indicators of neuronal activity. J Neurosci Methods...information on NIR FPs, their properties and applications. At the end, there is a Table that summarizes...