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Showing: 561 - 580 of 1036 results
  1. Antibody Guide

    Type
    Guide
    ...heavy chain constant region” or Fc. Different isotypes play different roles in the immune response, and ...of colors activated by different wavelengths, allowing for easy differentiation between multiple antibodies...roughly fifty different fluorophore colors in flow cytometry, but the largest panels used currently are in the...Antibodies are typically produced in one of three different methods: in animals, in cultured immune cells,...contain two Fab fragments, each recognizing a different epitope, bound by short peptide linkers. Diabodies...used as an antimicrobial agent. Other chemical interferences with conjugating reactions are possible. It ...chemiluminescent reaction in the presence of several different substrates. The luminescence produced by the reaction...
  2. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    ...reduced-in-size options often have reduced binding strength, half-lives, and/or specificity as well. They’...bispecific, meaning that each scFv can target a different antigen; binding of each can be either simultaneous...convert anyone to downsize their affinity reagents, trendy as that may sound. Validated antibodies are still... a scientist looking for that perfect fit!  References and Resources Additional resources on the Addgene...AddgeneBrowse our recombinant antibody plasmid collection References Grumezescu, A. M. (2018). Drug targeting and ...
  3. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...protein expression journey! Resources and References References Nabet et al. The dTAG system for immediate...expression of essential proteins (when knockouts aren’t viable). siRNA or RNAi are small, double stranded...days. There’s also a CRISPR for that! CRISPR interference (CRISPRi) targets a catalytically dead Cas9 ..., and expression is tunable by titration with different amounts of tetracycline. Tetracycline regulation...
  4. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    ...1013 - 1014 vg/kg, a dosage comparable to those currently used in human gene therapy trials. They further...reproducibility as scientists can assemble parts from different labs and experimental systems in their own research...mutants, determined protein expression levels from different promoters, and inhibited gene expression using...C-terminus of Cre (CreC) and the N-terminus of Cre (CreN) are fused to the truncated Phytochrome B (PhyBΔ...eye, muscle and epithelial cells, as well as in different stages of development, ensuring the breadth and...
  5. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    ...dependence on excitation power is important since different fluorescence imaging strategies use widely varying..., TIRF or spinning disk confocal use entirely different excitation power regimes. Therefore, it is unpredictable...the best way to compare the photostability of different fluorescent proteins? Measuring photostability...excitation power. By repeating these measurements for different fluorescent proteins and by comparing how the ...measurements reported in the literature are performed in different ways. We describe a number of issues with some...
  6. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    ... can only carry around 4.5 kb. Different AAV serotypes have preferences for specific cell types. For example...end of an experiment series would behave very differently (distracted) from the ones tested at the beginning...Experiment Behavioral scoring systems (experimenter differences in behavior observation); experimenter blinding...viral service for their behavioral studies. References Stoica L, Ahmed SS, Gao G, Sena-Esteves M. Gene...
  7. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    ...whenever you hear it (whether at a scientific conference or a karaoke bar), you probably want to sigh....methyltransferase 1 (DNMT1) to copy methylation marks from parental to daughter strands of DNA. When dCas9 is bound...nearly 100% demethylation without the confounds inherent to dCas9-TET, allowing one to study the causal... CRISPR-based tools for epigenetic editing.   References and resources:  Additional Resources on the Addgene...Addgene.org Read our CRISPR Guide to Epigenetics References Charlton J, Jung EJ, Mattei AL et al. (2020) ...
  8. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    ...Emily P. Bentley in October 2024. References and Resources References Demanèche, S., Bertolla, F., Buret...functions of natural plasmids. Created with BioRender.com. Resistance (R) plasmids: microbial defense... also causes outbreaks of food poisoning, the difference is virulence plasmids! This category includes...Mclellan, S. L., Willis, A. D., Comstock, L. E., & Eren, A. M. (2024). A cryptic plasmid is among the most...
  9. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    ...mammalian plasmid here and edit on! References and Resources References Kwart, D., Paquet, D., Teo, S. et...sequence constraints, or at least has looser or different requirements. Luckily, there are some available...necessarily more flexible than NGG; they’re often just different (e.g. NGAG, NGAN, etc.). Additionally, alternative...al). Additionally, SpG Cas9 with the same PAM preference of NG was isolated by the Kleinstiver Lab (read...
  10. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...28032891. This is a different version of Split-BioID. BirA* is fragmented at a different site than in Schopp... This can artificially select against PPIs that aren't stable outside the native cell context. The Split-BioID...of the BirA* fusion matters! Schopp et al saw different levels of fusion protein expression and biotinylation...research? Plasmids are available from Addgene! References Schopp, I.M., Ramirez, C.C., Debeljak, J., Kreibich...
  11. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ...or undetectable FRET, well-characterized FRET reference standards can be used to validate FRET measurements...sophisticated questions about cellular processes. Currently, FRET experiments can probe protein-protein interactions...Borghi 2016) to a selected protein. Overcoming current limitations to permit the use of multiple FRET ... diseases and even assist in drug discovery. References 1. Baird, Geoffrey S., et al. "Circular permutation...biomolecular structure and dynamics in live cells." Current opinion in structural biology 34 (2015): 52-59....
  12. Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux

    Type
    Blog Post
    ... to address the unique needs of students from different departments across the university and alterations...track postdocs nationwide. On this level, FoR is currently compiling a database of institutional responses...implementation of new strategies designed to cope with current increases. These two rulings along with progress...Howell -Founder of Howell Communications Philip Brenner, PhD - Assistant Professor of Sociology. University...public policy. Follow David on Twitter @driglar.   References: 1. Alberts, Bruce, et al. "Rescuing US biomedical...
  13. The Materials Science of Optogenetics Experiments

    Type
    Blog Post
    ...Several protocols have been written outlining the different steps and materials required for each [1, 2]. ...discussed above, each opsin variant has slightly different activation/inactivation kinetics. For example,...Biology from the University of Michigan and he is currently a Postdoctoral Fellow in the lab of Mary Jeanne... or follow him on Twitter @derekpsimonphd.    References 1. Gutierrez DV, et al. Optogenetic control of...channelrhodopsin for activation of CNS neurons. Current protocols in neuroscience / editorial board, Jacqueline...
  14. Chemogenetics Guide

    Type
    Guide
    ...Ligand(s) Effect Outcome (in neurons) Reference PSAM4 Gly Varenicline, uPSEM 792, uPSEM 817 Cl - influx Neuronal...RASSLs DREADDs PSAMs LMOs Plan Your Experiment References Chemogenetic tools are actuators for specific... of chemogenetic receptors. There are several different types of DREADDs that can be broadly classified...releases intracellular calcium stores. There are currently three Gq DREADDs based on human muscarinic receptors...cyclase and downstream cAMP production. There are currently two Gi DREADDs based on human muscarinic receptors...increasing intracellular cAMP concentrations. There is currently one Gs-DREADD, rM3D, that was created by replacing...intracellular region of a turkey erythrocyte β-adrenergic receptor with a rat M3 muscarinic receptor. This...
  15. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...randomized with a defined set of different trinucleotides (corresponding to different amino acids) in the regions...this, they injected the spike protein from two different coronaviruses into a llama over several days and...your target protein. The Seeger lab designed 3 different mRNA sybody libraries for use in the protocol,.... Find the Seeger lab's SARS-CoV-2 sybodies! References Geertsma ER (2013) FX Cloning: A Versatile High-Throughput...
  16. Magnetic Control of Proteins: More than a Dream

    Type
    Blog Post
    ...Find the MagLOV plasmid here!   References and resources References Duret, G., Polali, S., Anderson,...just be an artifact and eliminate sources of interference, but the result persisted. (Even better, multiple...be developed from other fluorescent proteins.  Currently, the team is working to identify how MagLOV’s ... of many optogenetic tools. And, since MagLOV’s parent protein AsLOV2 has already been adapted into a ...
  17. Antibodies 101: Flow Compensation

    Type
    Blog Post
    ...However, if your flow cytometer reads them out on different lasers, you should not need to compensate between... Guide Chart.  Pro tip! Some antibodies go by different names depending on your flow cytometer’s setup...look at the other channels you’re using first. A different fluorophore channel may be affecting your populations...first before you apply compensation values to your current channels. A notable exception If you have a sample...CD8+ T cells and cancer immunotherapies. She is currently a research technician in Arlene Sharpe's lab at...
  18. Twenty Years of Addgene Sharing: CRISPR

    Type
    Blog Post
    ...previous Addgene CRISPR publications.  References and resources References Pyhtila, B., Kasowitz, S., Leeson...with CRISPR pooled libraries, although we used different categories (Figure 2C). Note that in Figure 2,...through the years.  Libraries are divided into different categories based on function: knockout (KO), activate...have CRISPR plasmids deposited from labs in ~40 different countries, and we’ve shipped CRISPR plasmids to...
  19. Of Myc and Men

    Type
    Blog Post
    ...antibodies, context is key.    References and Resources References Evan GI, Lewis GK, Ramsay G, Bishop...cell proliferation and tumorigenesis.  Using two different synthetic peptides corresponding to linear sequences... or the C-terminus, Evan, et al. isolated six different monoclonal antibodies (Evan, et al. 1985). Three...-9E10 variable regions and sticking them onto different scaffolds. But over the years, several alternative...
  20. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    ...pathway engineering and optimization. However, current best practices for assemblies of more than 10 modules...rely on two-step hierarchical approaches using different Type IIS restriction enzyme specificities at each...Assembly test systems Our research focused on three different levels of assembly: High efficiency and accurate...products to market, New England Biolabs, Inc.       References 1. Vladimir, Potapov, et al. "Optimization of...cloning protocols Check out our molecular biology reference pages ...
Showing: 561 - 580 of 1036 results