Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 1,074 results for: CAN

Showing: 181 - 200 of 1074 results

  1. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    ...biotinylated proteins can be captured on streptavidin affinity beads or matrices. The bait protein can even target...2018). BioID and BioID2 can take hours to label proteins and APEX2, although it can rapidly label proteins...the mgSrtA variant can be displayed on the surface of a cell of interest, where it can label interacting...affinity of biotin for avidin, proximity labelling can capture weak/transient protein interactions as well...used for both soluble and insoluble proteins, which can be a challenge for classic antibody based affinity...methods.  BioID: a promiscuous biotin ligase that can efficiently label neighboring proteins The first ...proteins. Thus to capture protein complexes, scientists can simply express a bait protein fused to BirA* in cells...

  2. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    ... This step can involve multiple smaller steps that all help to ensure that antibodies can access their... that are not exclusive to IF, but they can have a significant impact on the outcome of your IF experiment...bind to their targets, most commonly proteins. You can then use fluorescent microscopy to look for those...ICC are essentially just more general terms that can also refer to assays using non-fluorescent visualization...polyclonal, which means that multiple secondary molecules can bind to a single primary, resulting in signal amplification...of possible primary antibodies that exist and you can begin to see why generating fluorophore-conjugated...made in different species on a single sample, you can then use the corresponding secondary antibodies conjugated...

  3. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...mutagenesis is a highly versatile technique that can be used to introduce specific nucleotide substitutions...or deletions) in a tailored manner. The approach can be used in conventional cloning (to introduce or ...contains the template, site-directed mutagenesis can be used to mutate the PAM sequence (an NGG sequence...Site Directed Mutagenesis In brief, point-mutations can be introduced to plasmids using primers (with the...plasmid containing the mutation). Similar results can be obtained with PCR screening using the primers ...repetitive sequences, but if present, a minor extension can usually ensure that the 3’-base(s) do not form secondary...larger constructs, but plasmids as large as ~6 kb can be amplified fairly easily by simply following the...

  4. The Addgene Antibody Data Hub

    Type
    Blog Post
    ...when appropriate, and can be saved at any point throughout the process, so you can work through it at your... antibody, the Data Hub can help you decide if an antibody is a good candidate for your experiment. If...data on which might work in your application. It can be quite difficult to decide on which antibody to...’s Antibody Data Hub Addgene’s Antibody Data Hub can help with this decision. In the open-access Data ...researchers interested in the Addgene antibody collection can look at user-deposited data detailing how an antibody...of the Addgene Data Hub.   Antibody data reports can be searched and sorted by gene/target, name, application... conditions in which the data was generated, you can scroll down the page to the materials and methods...

  5. Lab Automation at Addgene

    Type
    Blog Post
    ...one pod can pick up 96 tips for easy sample transfer between 96 well plates. The second pod can pick up...simple steps you can take to automate processes in your own lab. Some things your lab can do to get into...to do to find said robot and to find out how you can get one. As a starting point, I would recommend conferences...is it’s customizability. It has a large deck that can house many different types of devices. As long as...’s easy to pipette samples in and out of it, you can set up a procedure for it using the Biomek FxP. For... is pretty simple. By clicking a few buttons you can quickly tell the Biomek FxP how much sample to pick... making it easy to spread out your samples. This can be useful if you’re transferring samples from 96 ...

  6. How-to: 5 Steps to a Great Panel Discussion

    Type
    Blog Post
    ... They can result in lively discussions with both panelists and audience fully engaged. They can also be..., but the side can work. Only put out enough seats for the expected attendees (you can have extras ready...and program organizers can help make sure the event stays on schedule. People can only sit so long. I recommend...audience playing Candy Crush or reading email). My advice is always "don't do it unless you can do it well"... food or a fantastic view (these are nice if you can afford them). I am talking about making sure the ...interesting stories, riffs or opinions that each panelist can contribute. The moderator should identify the funny...panelists of the stories or topics you know they can speak about. There should be some spontaneity, but...

  7. 25 Work From Home Tips for Scientists

    Type
    Blog Post
    ...things you can do to prepare. Adapt to WFH while parenting Keep your kids on a schedule so you can rely ...that you can adapt to your kids' needs. Plan ahead. Think about and set up activities that kids can do more... week to flatten the curve, but working remotely can be more difficult if your work is based in the lab...outdoors is so huge, these are activities that you can easily do while staying away from others.  Meditate...assist in labs that are testing for COVID-19. You can also give back by supporting local businesses (ex...until now.  Network. There’s a lot of networking you can do online. See if a scientist or someone you’re connected...writing a full paper, that’s great. Otherwise, you can begin working on these drafts and for a few model...

  8. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    ...interspaced with linkers that can fold into a secondary structure that can bind specific proteins via aptamer...CRISPRi system can be repurposed to include modular RNA scaffolds. These RNA scaffolds can recruit various... to include modular synthetic RNA scaffolds that can recruit various transcriptional regulators (Zalatan...expression, multiple scRNA transcriptional programs can be regulated with dCas9 as a master ON/OFF switch...module, alternative sets of transcriptional programs can be layered with distinct programs controlled by each...expression at the targeted site. Multiple scaffold RNAs can be designed to allow simultaneous, multidirectional...tunable control of gene expression. This strategy can be applied to effectively change metabolic flux. ...

  9. Plasmids 101: Biotinylation

    Type
    Blog Post
    ...free avidin can be added to your sample. Any free biotin-binding sites on this complex can bind to your...biotin is added to a protein or macromolecule. Biotin can be added enzymatically or chemically. Enzymatic biotinylation... in the egg white. Avidin is a glycoprotein that can bind four molecules of biotin.   In the late ...glycoprotein that consists of four identical subunits that can each bind one molecule of biotin. Biotin and avidin...added to. Biotin also has an exposed side chain that can be easily manipulated to make reactive species without...polypeptide is biotinylated (more on this below) it can be incubated or co-expressed with avidin or a structurally...the 1960s, strepavidin. The Biotin-avidin complex can then be used and/or manipulated for a scientist’s...

  10. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    ...enough that their bulk can be a barrier at times. Smaller, slimmer affinity reagents can have deeper penetration..., meaning that each scFv can target a different antigen; binding of each can be either simultaneous or...kD but can have weak signals due to their monovalent nature. They are stable up to 80°C and can refold...readily available for a large number of targets, and can require specific conjugates. And, of course, once...often used in clinical applications. Fab fragments can be created in lab in situations where a suitable ...orientation of the two fragments. However, rigidity can be introduced through mutation, allowing diabodies...repeats, have high specificity and binding affinity, can be produced in E. coli, and are both soluble and ...

  11. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ...collection can be used for in vivo phenotypic screening of small-molecule libraries and can be combined...monogenic diseases. Additionally, retroviral vectors can regulate gene expression by delivering shRNA or miRNA...and the 2nd and 3rd Generation Lentiviral Systems can be found in our Lentiviral guide. Popular packaging...RNA-mediated gene silencing mechanism. Scientists can express siRNAs to direct the cellular RNAi machinery...Cre-recombinase. The Cre-lox system is a technology that can be used to induce site-specific recombination events...gene of interest pLenti-puro (Plasmid #39481) You can clone your gene of interest into this tetracycline...lentiviral vector backbone from the Shih lab. You can find more lentiviral expression vectors (empty backbones...

  12. Antibodies 101: Conjugation

    Type
    Blog Post
    ... streptavidin, both of which can bind up to four biotin molecules and can themselves be linked to a labeling... sodium azide, which can interfere with conjugation reactions. If needed, you can often remove the incompatible...spectrum, so they are great for multiplexing and can be used in a variety of assay types. Be sure to check...antibody conjugates in an earlier post, where you can find some additional tips for working with these ... which is light produced by a chemical reaction, can be detected using specialized imaging instruments...mass cytometry (Bendall, et al., 2012). And, you can also conjugate antibodies directly to beads for purification... linked to your antibody. This chemical coupling can be achieved through several different mechanisms....

  13. Career Coaching for Scientists: Why and Where Do I Find One?

    Type
    Blog Post
    ...for clear information on costs. Can you suggest some coaches? Yes I can. Please note that Addgene does ... many scientists.  Sometimes it can be helpful having a coach that can offer extra support for issues ...Professional Career Coaching can be an excellent tactic for scientists making a career shift or who wish...wish to improve their current job situation. What can a coach do that is different from an adviser, boss...accessible. Sometimes only a few hours of coaching time can make an enormous difference so don’t assume that ...offices, postdoc offices or professional organizations can provide referrals. There are bad coaches out there... a good one that is a style match for your needs can take some research.  It helps to spend some time ...

  14. Technical Design of a Western Blot

    Type
    Blog Post
    ...that can be done.  Note that each type of transfer requires different equipment. Wet transfers can be done... and can help amplify a weak signal. However, they have high cross-reactivity, vary significantly from...light-insensitive. They can detect femtogram levels of protein. These enzymes cannot be used for multiplexing...? What membrane? What should you block with? It can be so overwhelming that you might just stick with...monoclonal Both polyclonal and monoclonal antibodies can work well for a western blot. Polyclonal antibodies...batch-to-batch variation. Confirmation of sequence can be done from relatively cheap plasmid sequencing....of different rAbs that target the same protein — can be used to reproduce the benefits of polyclonal antibodies...

  15. Selecting Your Plasmid Purification Kit

    Type
    Blog Post
    ...final eluate. Bead-based kits can be more flexible in this situation as you can simply add more beads to your...Some applications require very little DNA and you can get away with slightly lower quality preps, while...Cloning Grade DNA (high quality/low quantity) – You can usually complete a cloning experiment with a relatively... on the experiment), but issues with DNA quality can inhibit important reactions like PCR or restriction... on the experiment, mammalian cell transfections can require quite large amounts of DNA (easily in the...DNA quality? There are a few different issues that can arise during the plasmid purification process. One...possibility of damaging the plasmid DNA itself, which can happen if you leave the sample in a lysis buffer ...

  16. MXS Chaining

    Type
    Blog Post
    ...procedure can be used to incorporate each element into the final construct. This strategy can be used to... each other, a third module cannot be placed between them, but rather, can only be added to the 5’ or ...synthetic biology for example, high-throughput cloning can be used to combine the functions of different genetic...reporter constructs. These fluorescent reporters can be used to detect structure and protein localization...sequences, and more. By combining these components, one can generate constructs to visualize cell cycle dynamics...directional, meaning that (in the previous example) one can control whether the third module is added to the ...References Golden Gate Type II restriction enzymes Can assemble up to 10 modules in a single step. Through...

  17. Healthcare Consulting: A Door to the Business of Life Sciences

    Type
    Blog Post
    ...drugs, the diseases they can treat, and diagnostic advancements. Consultants can help these teams develop...of time can sometimes become cauldrons on the verge of boiling. On other occasions, clients can be excessively...are just examples, and healthcare consulting firms can be involved in many similar activities depending ...shifting goal posts” as a part of a case, consulting can sometimes be challenging. The fast paced nature of...working hours and the unpredictability of schedules can take on employee morale and productivity, and many... any highly collaborative work place, challenges can arise due to conflicting work styles, team situations...abruptly change needs and expectations. Such situations can get stressful and to help manage that, most consulting...

  18. The Challenges of Cell Culture

    Type
    Blog Post
    ...the line, and can include assays for authentication and contamination) and cell lines can become cross ... The exact number of passages can vary between cells lines, as this can be dependent on the mutation rate...approach, the many mistakes of the past can be prevented and we can progress faster to desired clinical endpoints...retracted and there is a loss of reputation that can impede collaborations, especially with industry. ...test known as Short Tandem Repeat (STR) profiling can be used to ID the cell line. The same test is so ...forensic technique (aka DNA fingerprinting) that can place suspects at the scene of a crime (every human...human in the world has a unique STR profile) and can also be used as a paternity test. The test is accurate...

  19. Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies

    Type
    Blog Post
    ...into the brain can usually be achieved by a directed brain injection, but some serotypes can cross the blood...-associated viral vectors (AAV), neuroscientists can combine classical genetic tools with the manipulation...defined cell types. Specific transgene expression can be also achieved in nontransgenic animals by using... viral strategies Combining different approaches can increase the specificity of cell targeting even further...further. For example, injecting AAVs at one location can result in expression of an optogenetic payload in...the brain. A more locally defined set of neurons can then be targeted with optical stimuli. The restricted...targeted cell population.  Combinatorial strategies can also be used for retrograde tracing: visualizing ...

  20. Searchable and Sortable gRNAs for Your Next CRISPR Experiment

    Type
    Blog Post
    ...searchable, which can help exclude the data that you are not interested in. You can use the carats in ... with CRISPR, using one of these validated gRNAs can save you the time that would be spent making and ...and testing entirely new gRNA designs. You can now easily find many validated gRNAs in our newly curated...sequence will yield a useful gRNA.  Alternatively, you can select a target sequence which has already been shown...therefore usually transcribe your gRNAs in vitro, you can send us the sequences of your effective gRNAs, and...sequences here. Features of the validated gRNA table You can access the datatable here. Current table columns ....  The data in any column are searchable and you can search by any part of the data. For example, entering...
Showing: 181 - 200 of 1074 results