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We narrowed to 987 results for: igh

Showing: 801 - 850 of 987 results

  1. dTAG - You're it!

    Type
    Blog Post
    ...mouse models. The dTAG system The dTAG system pairs highly selective small molecule FKBP12F36V degraders (...observed little to no toxicity with dTAG-13 at doses as high as 20 µM (see Figures 3 and 5 in Nabet et al., Nature...

  2. The Breast Cancer Microenvironment: A Tumor’s Backstage Team

    Type
    Blog Post
    ...microenvironments and what encircles a disease is highly prone to affect it. While tumors may be the stars...surroundings and being shaped by them. Another set of highly influential microenvironment cells are Cancer-Associated...

  3. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    ...amino acids. The use of radioactive materials is highly regulated, requires designated work areas and come...passionate about communicating science to all audiences. Right now, she is a science instructor at The Innovation...

  4. Plasmids 101: Modular Cloning

    Type
    Blog Post
    ...of large-scale assembly useful, you may ask? You might be surprised by how multi-purpose it can be. Let... Plant Parts Kit Resources on the Addgene blog High Complexity Golden Gate Assembly with NEB’s DAD Interview...

  7. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ...1948 as a variation of the more commonly observed light emission by fluorescence. The widespread use of ...distances that are too miniscule for conventional light microscopy. Don't FRET - Use these FRET resources...

  8. Antibodies 101: ELISA (Enzyme-linked Immunosorbent Assay)

    Type
    Blog Post
    ... is biotin-conjugated, streptavidin (which has a high affinity for binding biotin), is conjugated to the...interest. The optical density (OD) - the measure of light absorbed by a solution - of these wells is plotted...

  9. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    ...are a multitude of known AAV serotypes that have slight variations in their viral capsids. Novel forms ...AAV types, AAV1, 2, 3 and 5 are thought to be as high as 70% among the human population. These pre-existing...

  10. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    ...for these RNA sequences (3 nM), however, was quite high and above what would be expected in patient samples...daunting. It took a true team effort and many late nights, not unlike most research, but the urgent need ...

  11. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    ...fold (Davis et al., 2002). However, polybrene is highly sensitive to freeze-thaw cycles and should be stored...resistance marker expression following transfection, higher-level expression after approximately 72 hours is...

  12. Illuminating Epigenetics with A FRET Based Biosensor

    Type
    Blog Post
    ...expression can change as well. The more frequently and tightly bound DNA is to histones, the more compacted it...occurs at CpG islands (regions of the DNA with a high frequency of C and G nucleotides). The conversion...

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

    Type
    Blog Post
    ...my experience the best set-up is panelists in a slight semi-circle (so they can see each other) on comfortable...redirect the conversation. The person you cut off might be miffed but the other 200 audience members will...

  17. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...think I need. Unlike lentivirus, AAV requires a higher number of viral particles for efficient gene transfer...only harvest the cell pellet. My yield would be higher if I used PEG precipitation, but for my experiments...

  18. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    ...Gamma-retrovirus genome (left) and an HIV lentiviral genome (right.)  HIV seems an unlikely candidate for use as...genomes in one retroviral particle: pseudodiploidy and high rate of genetic recombination. Proceedings of the...

  19. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...bp, but pay attention to the sequence’s Tm, as a higher Tm give more efficient recombination. We normally... two colonies for simple cloning procedures. Use high fidelity DNA polymerase This will minimize random...

  21. A Guide to Starting Your Own Journal Club

    Type
    Blog Post
    ...wanted to think more closely about the wealth of high-throughput sequencing (HTS) data we generate and...the aim of the journal club is to begin building a high level understanding across multiple teams or to ...

  23. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...cumbersome to use and lead to lower viral titers due to higher plasmid number. For more information on the key...Lentiviral particles with the VSV-G envelope have high infectivity in a wide range of cell types, so they...

  24. A Career in Grants: How To Become a Grant Professional

    Type
    Blog Post
    ...professionals identify grant opportunities with the highest likelihood of funding and develop a strategy and... project management, and program evaluation are highly transferable. Use your experience and your passion...

  26. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ...eukaryotic ribosome. One resistance gene on your vector might require two different selection molecules! It all...protein, to visualize vector uptake. These reporters might be used as markers for automated cell sorting or...

  27. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    ...Although AYBE, AXBE, and ACBE do not approach the high product purity of CBEs and ABEs, they could be useful...transversion editors, this editor had relatively high indel rates ranging from 4.7% to 30% at tested sites...

  28. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    ...which help a researcher understand if a material is right for their experiment.     Table 1: Most popular ...learned to clone from the Addgene website.” We delight in seeing our resources shared via social media...

  29. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ...to Cas9, which requires that each DNA target have high sequence identity to the spacer sequence and contain...to detect single molecules of an RNA species with high specificity (Gootenberg et al., 2017). This system...

  33. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    ...? Let’s take a look at codon usage and when you might want to optimize codons for expression in a particular... on the codon usage frequency in a reference of highly expressed genes (Sharp and Li, 1987). In 2000 the...

  34. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    ... method, they found that SaCas9 did not display higher levels of off-target activity than SpCas9, confirming...editors. While using AAV to deliver large transgenes might seem daunting, it’s actually a challenge that the...

  35. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    ...This post was contributed by Kutubuddin Molla, a Fulbright Visiting Scholar at the Pennsylvania State University...Cuttack, Odisha, India. Currently Kutubuddin is a Fulbright Visiting Scholar at the Pennsylvania State University...

  37. Bioinformatics at Addgene

    Type
    Blog Post
    ... was to ensure the scientific community obtained high-quality data for our DNA-based reagents. Not only..., one of our first problems was deciding on the right system for us to start using. Prior experimenting...

  38. Plasmids 101: Control Plasmids

    Type
    Blog Post
    ... you are trying to repress a gene, your control might be a system where expression of that gene is knocked...selection of the positive control plasmids should be highly specific to the experiment and the interrogation...

  39. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...ionizing radiation, chemotherapeutics, and ultraviolet light. In this post, we will describe the mechanism of...systems, ssODN templates generally provide the highest frequency of HDR edits.  Check out our blog post...

  41. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ...,000 phosphorylation events in the proteomes of higher mammals (2). How can we begin to understand the...intersection of cell signaling, metabolism, and the mighty mitochondrion. You can follow her on Twitter @nieminm...

  42. How to Negotiate a Successful Lab Start Up Budget

    Type
    Blog Post
    ...financial aspects. Money often carries an emotional weight. Therefore, too much focus on money can dissuade...point, your budget bottom line costs may be rather high. That’s normal, because this is where you can start...

  43. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    ...components of CGBEs. Their eOPTI-CGBE and cOPTI-CGBE had higher editing efficiency and produced fewer indels than...accessible. It's not entirely clear why this is, but it might point to a more dynamic picture of chromatin than...

  44. CRISPR Library Amplification

    Type
    Protocol
    ...LB and weigh pellet. The total weight of each pellet should be ~1-2 g. Pro-Tip Make sure to weigh the empty... plates at 30 ℃ overnight. Plate 2.5 mL of the transformed cells on each of the eight bioassay plates ...are often used for screening, barcoding, or other high throughput multiplexed experiments. These pooled... we recommend the use of a diagnostic digest and high-throughput next generation sequencing (NGS). Select...Workflow Timeline Day 1: Transform, recover, set up overnight growth (Estimated time 2-3 hours) Transformation... Alternatives include Stbl4 cells or other ultra-high efficiency electrocompetent cells that are suitable...of electrocompetent cells is essential to ensure high efficiency uptake of plasmid library DNA. This quantity...

  45. Protocol - How to Inoculate a Bacterial Culture

    Type
    Protocol
    ...numbers, 50 or more per cell and have a high copy number. High copy number plasmids should only need to...determine if your plasmid is high or low copy. I didn't get any growth after overnight incubation. What went ...However, a liquid culture is capable of supporting a higher density of bacteria and is used to grow up sufficient... The following protocol is for inoculating an overnight culture of liquid LB with bacteria. Video Watch...Prepare liquid LB. For example, to make 400 mL of LB, weigh out the following into a 500 mL glass bottle: 4 ...2 mL in a falcon tube, but for larger preps you might want to use as much as a liter of LB in a 2 L Erlenmeyer... sterile aluminum foil or a cap that is not air tight. Incubate bacterial culture at 37°C for 12-18 hr...

  46. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...Introduction Many molecular biology techniques require highly purified and concentrated plasmid DNA. This page...microcentrifuge Desktop vortexer Vacuum (optional) Reagents Overnight culture of bacteria transformed with your plasmid...Protocol: Generalized DNA Purification Grow an overnight culture of bacteria . Pro-Tip Refer to appropriate... with DNA preparation. Pro-Tip If your entire overnight culture cannot fit into a single centrifuge tube...of dH 2 O Store Solution III at 4°C Grow 2 mL overnight cultures from single colonies of bacteria containing...often thought that an incubation of 20 min to overnight at -20 °C or -80 °C will improve precipitation... the tube for 5 min at room temperature on the highest setting. Note: You should see clearly separated...

  47. Pouring LB Agar Plates

    Type
    Protocol
    ...least 30 min. The high pressure will prevent your gel mix from boiling over at high temperature. Pro-Tip...your agar is at the right temperature, we recommend using a laser thermometer. Light the flame at the plate...them out at room temperature overnight to allow them to dry. After overnight drying, we place the plates...the appropriate liquid solvent. See table to the right for appropriate antibiotic concentrations. Antibiotic...its cap or aluminum foil (but do not make an air-tight seal!) and tape the bottle with autoclave tape. ... from the autoclave and will cool your gel-mix slightly. Even so, you should always use thermally insulated...resistant to the antibiotic. Incubate both plates overnight at the appropriate growth temperature and check...

  48. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...protein, Apolipoprotein L3 (red) overnight, blocked, and incubated overnight with Anti-Desmin [D7]. Plates...protocol may need to be adjusted to accommodate slight differences between products. Addgene does not ...plate seal and incubate at 37 °C for 30 min , or overnight at 4 °C . Section 2: Block the plate Prepare the...set to 400 rpm for 2 h at room temperature or overnight at 4 °C . Section 3: Primary antibody incubation...set to 400 rpm for 2 h at room temperature or overnight at 4 °C . Section 4: Secondary antibody incubation...set to 400 rpm for 2 h at room temperature or overnight at 4 °C . Section 5: TMB reaction Carefully remove...to blue and generally takes between 15 to 30 min. High concentrations of HRP yield a greenish solution....

  49. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...Protocol Version 1.0. December 2006. Copyright Addgene 2006, All Rights Reserved. This protocol is provided...ligation, use only long-wavelength UV light. Short wavelength UV light will increase the chance of damaging...culture plate. Incubate cells at 37°C, 5% CO 2 overnight. Although cells should regularly be passaged in... in ten 6 cm plates and grow at 37°C, 5% CO 2 overnight. Day 2: b. The target cells should be approximately...Plate target cells and incubate at 37°C, 5% CO 2 overnight. Day 2: b. Target cells should be approximately...plate, add between 0.05-1 mL virus (add 0.5 mL for a high MOI, and 0.1 mL for a low MOI). Scale the amount...cell line. d. Incubate cells at 37°C, 5% CO 2 overnight. Day 3: e. Change to fresh media 24 hours after...

  50. Lentivirus Production

    Type
    Protocol
    ...PEI μg ratios provided high transfection efficiencies as measured by the highest proportion of GFP positive...without limiting cell growth. Left panels: bright field images; right panels: GFP channel images. Last reviewed...meter Stir plate Magnetic Stir Bar Reagents DMEM high glucose, Corning 10-013-CV L-alanyl-L-glutamine ...alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat-inactivated FBS and ...the packaging cell line is critical for obtaining high viral titer. 293T cells should be split 3 times ...purification should include an endotoxin removal step. For high quality plasmid DNA, the plasmids should also be...transfection to determine what ratio gives the highest percentage of GFP positive cells. Refer to the ...
Showing: 801 - 850 of 987 results