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We narrowed to 545 results for: SEC

Showing: 61 - 80 of 545 results

  1. Antibodies 101: Chimeric Antibodies

    Type
    Blog Post
    ...anti-rat IgG1 secondary to detect Protein A and green fluorophore-tagged anti-mouse IgG2a secondary to detect...IgG1 secondary antibody to bind to the anti-Protein A antibody and an anti-mouse IgG2a secondary antibody...constraining using the indirect method since a secondary antibody reactive to a certain isotype will bind...antibodies that are the same isotype then the secondary antibody will bind to both and you will not be...the same isotype and will react with the same secondary. If you were to change one of the antibody’s isotypes...isotypes, however, you could then use 2 different secondaries for your antibody pair and visualize both targets...multiplexed indirect staining methods since the secondary antibody used will react to both primary antibodies...

  2. Which Fluorescence Microscopy Techniques is Best for Me?

    Type
    Blog Post
    ...microscopy technique to incorporate optical sectioning. Optical sectioning refers to the ability to extract light...) tissue sections and 3D cultures These samples do not necessarily require optical sectioning, but open...this, the use of multiple fluorophores can add a second layer of contrast, color contrast, to an image ...volumes to enhance spatial contrast (i.e. optical sectioning (2) or super-resolution (3). Fluorescence microscopy...establish for this to happen.Using red light has a second advantage: red light penetrates deeper into biological... on the sample, a temporal resolution on the millisecond scale may be needed. Most commonly these experiments... of the coverslip (4). Essentially, an optical section is achieved that boosts contrast by filtering out...

  3. Plasmids 101: Protein Expression

    Type
    Blog Post
    ...conditions compared to other systems. Insect expression systems Insect cells can also be used to produce ...are two types of insect expression systems; baculovirus infected and non-lytic insect cells. Baculovirus...genes integrated into the insect cell genome. Both of these types of insect expression systems can be ...stable proteins. Protein expression refers to the second step of this process: the synthesis of proteins...production and purification. These include mammalian, insect, bacterial, plant, yeast and cell free expression...halts protein production, but there are non-lytic insect cell expression systems (sf9, Sf21, Hi-5 cells)...of large amounts of protein. Some drawbacks of insect cell expression systems include that virus production...

  4. A Guide to Designing a Scientific Poster: Content, Layout, and More

    Type
    Blog Post
    ...acknowledgements sections. Some other additions you might wish to include are a references section, contact information...styles; one for your section titles, and a different one for the body of those sections. This strategy helps...poster. While a poster generally contains the same sections as a primary research article, it’s important ...common poster size, but this can vary as well.  Sections of your poster In almost all scientific posters...creative use of a footer in your poster. This is a section where you can insert your references and contact...font here to be much smaller than in other poster sections, so you can include a lot more. Font size: Making...helps your sections stand out, and makes the entire poster more navigable.  Choose your content wisely...

  5. Antibodies 101: Selecting the Right Antibody

    Type
    Blog Post
    ...in a similar way as you plan to. To secondary or not to secondary?  The application will also determine...primary-conjugated) or indirect (secondary) methods. In those cases, remember that a secondary should be used when ...substrate, such as a fluorophore, or a conjugated secondary antibody that will bind to your primary antibody... the precise location of a protein in a tissue section then a directly-conjugated antibody will provide...immune complexes are smaller than a primary:secondaries:conjugates complex. If you are trying to visualize...protein in low abundance on a Western blot, a secondary antibody is ideal as it will help to amplify the...you can choose different fluorophore-conjugated secondary antibodies specific to each isotype or species...

  6. Antibodies 101: Fab Fragments

    Type
    Blog Post
    ...fragments make good secondaries for cases where you need to pre-form primary-secondary antibody complexes...species as your secondary antibody to your sample prior to adding your primary and secondary antibodies, ... lab, this is the region that determines what secondary antibody you should use when performing indirect...portion means that most of the common anti-IgG secondary antibodies you already have in your refrigerator...or you need a light-chain specific conjugated secondary antibody. These requirements limit the flexibility...contrast, anti-IgG antibodies, commonly used as secondary antibodies, have much broader applicability. It...as when performing IHC in a sample where your secondary antibody recognizes endogenous antigens. In these...

  7. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    ...may have an entirely different use in mind! This section of our plasmid pages is relatively new, so some...without a purpose and would like to fill in this section, let us know! Is there a way for me to quickly... these are found in the "Resource Information" section of the plasmid page (discussed below). Great! ...use in your Materials and Methods and References sections when citing the use of your colleague’s plasmid...information about the plasmid contents? The middle section of our plasmid pages contain all the detailed information...backbone, and each is given its own gene/insert section on the plasmid page. Tags are listed according ... successful cloning. Cloning information: This section provides useful information about how the vector...

  8. Building and Validating Recombinant Antibodies: A Case Study

    Type
    Blog Post
    ...Benefits and pitfalls of secondary antibodies: why choosing the right secondary is of primary importance...multiple subclasses of IgG antibody. Broad range secondary antibodies are commonly used in research laboratories... the different subclasses, but there are also secondary antibodies that are specific to each subclass....2012), which limits the use of subclass specific secondary antibodies to detect multiple targets with different...original hybridoma. By using subclass specific secondary antibodies and subclassed switched R-mAbs, multiple...Plasmid-Based Recombinant Antibodies  Plasmids 101: Secondary Nanobody Toolbox Antibodies 101: Introduction ...

  9. 5 Steps to Writing Grant Proposals for Grad Students

    Type
    Blog Post
    ... typical sections in a NIH RFA, for example, and tips to consider while writing each section. Research...the limitations of current methodologies. This section sets the stage for why your funded proposal will...of the text. Specific aims The Specific Aims section highlights each milestone that must be met in order...Typical proposals consist of 3 to 5 aims, with secondary milestones to meet along the way to achieving ...detail (within the page limits) provided in this section will convince the reviewers that the proposal is... a first time budget writer, you may find this section challenging because it’s hard to determine the ...

  10. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...with CRISPR-Cas9. The survey had two sections, the first section asked which of the two methods researchers...researchers found to be more successful, and the second section asked basic questions about the NgAgo protocols...unprecedented freedom to target any sequence of DNA. Second, NgAgo target specificity is dictated by a DNA ...Blog - Leave suggestions and tips in the comments section below or contact [email protected] if you'd like...community on NgAgo forums, blogs, or in the comments section below. Keep pipetting and happy genome editing!...

  11. Addgene-Seeding Labs Plasmid Grant: Accelerating Science Globally - 5 Scientists, 4 Countries & 3 Continents!

    Type
    Blog Post
    ... and analysing the insecticidal potential of Cry proteins in order to manage insect pests and to support...toxicity against local insect pests. By developing biological agents such as insecticidal proteins, the use...from the identification of plant compounds and insecticidal proteins to the development of renewable biotechnology... desaturating fatty acids. Identification of Insecticidal Proteins Linh Bao Ton (M. Sc.), Nong Lam University...

  12. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    ...assaying a secreted protein, first block the Golgi apparatus with Brefeldin A to inhibit secretion and then...binds to a target and a fluorochrome-conjugated secondary antibody binds to the primary antibody. Direct...non-specific staining that may arise from the use of secondary antibodies. It’s also particularly useful for ...beneficial when expression levels are low as multiple secondary antibodies can bind to a primary antibody thus... staining protocol, you should also include a secondary antibody control in which the primary antibody...to detect background staining arising from the secondary antibody.  It is also recommended that users include...

  13. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    ...the primary and secondary antibodies from your membrane, clearing the way for the second blot. There are...that permanently stain the membrane, making that section of the blot unusable for reprobing. Preparing your...simply repeat the imaging step to check if any secondary antibody remains, whether that involves adding...re-blocking your membrane and re-incubating with your secondary antibody. This is the one time in western blotting...western blot as normal. Be sure to rinse away the secondary antibody you used to assess the stripping, and...

  14. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    ...will focus on these tools in this blog post). The second major area deals in post-experiment quality control...software has been designed for this purpose. This second area is extremely important and given the lack ...simple catalogue of reported sgRNA designs. The second database, CrisprGE (published 27 Jun 2015), is ...goals. Examples:  “feature aware”, “SNP aware”, “secondary structure aware”, and “microhomology aware”. Trends...language or another. Supported organisms: This section lists the genomes supported by each tool. The genomes...have currently available organisms listed in this section. CRISPR software advice for specific users You... pursued. You should also look at the glossary section of the CRISPR Software Matchmaker, it provides ...

  15. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    ... unclear how much they will actually care. The section of the book I’d really like to press upon my non-scientist...called simply “Bad Science.” Zimmer kicks off the section saying “science itself is never inherently bad ...call bad science.”  The three chapters in this section describe the challenges of trusting science in ...essential tool in biology also falls within this section. The GFP story has its own Franklin in Doug Prasher...eventually replace scientists themselves.  The sections on optogenetics and CRISPR of course address some...competitiveness of U.S. science and technology.” This last section of the book, Future Science, describes the author...

  16. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    ...plasmids, one encoding gag and pol and a second encoding rev.  Secondly, 3rd generation packaging systems have...pose too much of a biosafety risk to scientists. Second-generation vectors are likely sufficient for most...based on whether or not they have a chimeric 5'LTR. Second-generation transfer plasmids have a wildtype 5'... protein and consequently are considered safer. Second-generation transfer plasmids MUST be packaged with...Third-generation vectors contain gag, pol, and rev. Second-generation contains those three genes plus tat....

  17. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ...light-pulse at one wavelength and stay on until a second pulse of light, with a different wavelength, turns... specific neurons without needing to consider secondary signaling pathways. There are many different opsin...exposed to blue-light and activates downstream secondary signaling pathways. Photoswitchable tethered ligands...Created with BioRender.com.   Controlling secondary pathways with photoswitches OptoXRs are great ...Photoswitches can be used to activate or suppress secondary signaling pathways with light. This is achieved...Bamberg, E., Nagel, G., & Deisseroth, K. (2005). Millisecond-timescale, genetically targeted optical control...

  18. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    ...research well. Who could fill these different roles? A second or third mentor. Therefore, seek out additional...First, your main project may seem stuck in a rut. Second, many of the great discoveries in science arose...effectively tell a story? Generally, the answer to the second question is yes. As you get better at telling your...drafts, reviews, cover letters, fellowships, etc. Second, get constructive feedback from experts. If someone...mentor, manage, communicate, and teach The dirty secret about becoming a group leader is that people are... His interests include the quality control of secretory proteins in the Endoplasmic Reticulum, optimization...

  19. Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers

    Type
    Blog Post
    ...Cloning of the second plasmid can be done in one day in one assembly reaction. This second plasmid saves...CRISPR/Cas system, what we are doing is adding in a second plasmid to target an epitope tag onto the transcription...different assays, not just transcription factors. The second vector is a homology directed repair donor plasmid...selection marker. If things work correctly, the second plasmid inserts a small epitope tag called FLAG...

  20. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    ...bloggers Joachim Goedhart and Marieke Mastop from the Section of Molecular Cytology and Van Leeuwenhoek Centre... selecting a bright fluorescent protein. In the second post of this series, we will discuss how to select...the fluorescence intensity evolves over time. The second issue is that, in experiments that measure photostability...Joachim Goedhart is an assistant professor at the Section of Molecular Cytology and Van Leeuwenhoek Centre...joachimgoedhart.   Marieke Mastop is a PhD candidate at the Section of Molecular Cytology (University of Amsterdam)...
Showing: 61 - 80 of 545 results