We narrowed to 296 results for: SAM activators
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TypeBlog PostPublishedJuly 14, 2021, 1:15 p.m....recognize the same region, or epitope, of the target protein. These antibodies are also all the same isotype...heterogeneous mixture of many antibodies that recognize the same protein. In the immune system, antibodies are produced...peptide sequence derived from this protein. At the same time, an adjuvant such as KLH or Freund’s adjuvant... without causing high background due to cross-reactivity. When to use polyclonal antibodies One advantage...clones will still recognize your protein. For the same reason, they’re also useful in experiments where...experiment, you should either order several vials of the same lot of polyclonal antibody, or select a monoclonal...your polyclonal antibody, you’ll want to use the same positive control across experiments to be aware ...
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Antibodies 101: Chimeric Antibodies
TypeBlog PostPublishedApril 25, 2023, 1:15 p.m.... are the same isotype and therefore cannot be indirectly probed simultaneously on the sample. What to ...because the pair belong to the same isotype and will react with the same secondary. If you were to change...the indirect method since a secondary antibody reactive to a certain isotype will bind to all members ...that group. If you use two antibodies that are the same isotype then the secondary antibody will bind to... Figure 2: Two primary antibodies with the same isotype can not be used for multiplexed indirect...parental species which can be bound by broadly reactive secondary antibodies to that parental species.... -
Corralling and Cornering the Goldilocks Antibody
TypeBlog PostPublishedAug. 9, 2022, 1:15 p.m....compare different clones of the same target to see if the results are the same/similar, emphasizing the antibody...your sample to. For example, we use our negative control to compare exposure times of our samples. A prominent...confirming it will produce the same results even to a different sample. Finally, as a lab we discuss the...underlying causes of this unreliability: cross-reactivity (binds to non-target proteins), variability (...Baker, 2015), causing a huge waste of precious samples, time, and funds – up to US$350 million in 2015...noise. We then compare the negative control to a sample with the primary included to confirm that the signal...antibody and not from the background noise. If the sample images similarly to the negative control, that ... -
Adenoviral Vector Production and Troubleshooting
TypeBlog PostPublishedJune 14, 2018, 11:44 a.m....particles = number of particles per mL of sample. The purity of the sample can be estimated by looking at the...Adenoviral vectors (AdV) are attractive vectors for research applications and gene therapy: they can ...Trapnell. "Evaluation of the concentration and bioactivity of adenovirus vectors for gene therapy." Journal... -
Viral Vectors 101: Producing Your rAAV
TypeBlog PostPublishedJuly 16, 2024, 1:15 p.m....known titer on every qPCR run. Note that the rAAV sample has to be digested with DNase before undergoing...Grimm, D., Kern, A., Pawlita, M., Ferrari, F., Samulski, R., & Kleinschmidt, J. (1999). Titration of AAV...Harvey, B. K. (2017). Assaying the Stability and Inactivation of AAV Serotype 1 Vectors. Human Gene Therapy..., J., Hennigs, J. K., & Körbelin, J. (2021). Inactivation of Adeno-Associated Viral Vectors by Oxidant-Based... -
Antibodies 101: The Basics of Western Blotting
TypeBlog PostPublishedMarch 8, 2021, 2:41 p.m....thiols, to cleave disulfide bonds. Load the samples, one sample per lane, onto the top of a resolving gel...all of the proteins in a sample migrate through the stacking gel at the same pace and enter the resolving...consistent size and density present in all samples. Uneven sample loading or incomplete protein transfer ...Figure 1: Overview of the Western blot process. The sample, a mixture of protein is separated in a gel, transferred...is the first step of a western. To prepare the samples for SDS-PAGE, measure the protein content and normalize...normalize to ensure equivalent loading. Denature the samples to their primary amino acid sequence by boiling...crosslinked polymer acrylamide. In addition to the samples, a protein standard of known molecular weight in... -
Antibodies 101: Validation
TypeBlog PostPublishedMarch 24, 2022, 1:15 p.m....antibody signal in the retinal sample, but little to no signal in the liver sample, which you do (C), so your...epitopes on the same target reduces the likelihood that both antibodies would bind to the same off-target ...core goals of antibody validation: Confirm reactivity with the target antigen - Recognition of an immunogen...to - particularly when you start using complex samples such as whole cell lysate or tissue sections. It...require variable expression of your target across samples - it is hard to distinguish between specific, invariable...there is a wealth of transcriptomic data from this sample type, so you use your bioinformatics skills (or...Removing (or reducing) the protein of interest in your sample, should lead to reduced or eliminated antibody ... -
Viral Vectors 101: Pseudotyping
TypeBlog PostPublishedApril 20, 2021, 2:30 p.m....pseudotype To pseudotype your viral vector, you use the same protocol for generating lentivirus, but you use ...Different pseudotyped viruses are differentially inactivated by nonspecific complement mechanisms in serum...anti-alphaGal antibodies. The VSV-G envelope protein is inactivated by serum regardless of the producer cell type...high concentrations as described above and is inactivated by the complement system in serum (DePolo et ...Vector Particles Produced in Human Cells Are Inactivated by Human Serum. Molecular Therapy 2:218–222 .... -
Cultivating Community Science at BosLab
TypeBlog PostPublishedNov. 2, 2017, 1:35 p.m.... DIY Biology and, on occasion, would meet in the same space where BosLab exists today. Back then, it consisted...saw the potential in the space and decided to reactivate it. We started out by holding a few journal club... getting the community to collect various soil samples, extract DNA, sequence it, and analyze the data... characterize the microbiomes of the different samples. The goal is to not only learn something about ... -
Plasmids 101: Knockout/Knock-In Plasmids
TypeBlog PostPublishedDec. 1, 2016, 3:30 p.m....strategies to investigate a gene's function is to inactivate, or "knockout", the gene by replacing it or disrupting...describe the techniques and the plasmids used to inactivate specific genes in mammalian cells. Despite the...one of its exons has simply been replaced with the same sequence flanked by LoxP sites while the resistance... are easier to use and are more efficient at inactivating genes. CRISPR can target a genomic sequence ... -
Inside Addgene's 10 Year Anniversary Gala
TypeBlog PostPublishedJuly 3, 2014, 3:48 p.m.... the interactive photo graffiti wall. I had a lot of fun playing the prototype of an interactive game ...playing all night was hand-picked (like our plasmid samples!) by Addgenies. If you were there, you might have... -
In Living Color: The Skinny on In Vivo Imaging Tools
TypeBlog PostPublishedMarch 27, 2014, 5:14 p.m....last year, Verkhusha added two far-red light photoactivatable (PA) near-infrared fluorescent proteins (FPs...signal-to-background ratio, thus, allowing higher resolution in samples containing substantial autofluorescence background...751-4. doi:10.1038/nmeth.2521) Far-red light photoactivatable near-infrared fluorescent proteins engineered... -
Plasmids 101: Antibiotic Resistance Genes
TypeBlog PostPublishedJan. 30, 2014, 3:29 p.m.... are trying to disrupt or delete, which both inactivates the gene and acts as a marker for the mutation...of experiments it is best practice not to use the same resistance cassette for the mutation and for plasmid...cassettes and your gene of interest is cloned into/inactivates or completely removes (in the case of Gateway... -
An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency
TypeBlog PostPublishedDec. 17, 2020, 1:30 p.m....approach – co-CRISPR from Craig Mello’s lab – used inactivation of the unc-22 gene as their selection marker...Drosophila S2 cells (Böttcher et al., 2014), inactivation of non-homologous end-joining results in a further...changes 5’ to the PAM, sequences corresponding to the same strand as the PAM produced a higher editing efficiency...elegansGenome with Lethal Mutation Co-Conversion and Inactivation of NHEJ Repair. Genetics 199:363–377 . https... -
Improving Transduction Efficiency with Exosome AAVs
TypeBlog PostPublishedMarch 14, 2019, 1:54 p.m.... have a higher transduction efficiency, i.e. the same number of AAV particles is able to infect more cells... protects them from antibody recognition and inactivation (György et al., 2014), but scientists have yet...difficult to reach, and are more resistant to inactivation through anti-AAV antibodies. References György... -
Visualizing Protein Turnover In Situ
TypeBlog PostPublishedJan. 16, 2018, 3:20 p.m....classic approach for studying protein turnover is radioactive pulse-chase experiments. However, these experiments...require radiolabeled amino acids. The use of radioactive materials is highly regulated, requires designated... suspected to interact, are added to a cellular sample. Both antibodies have a short antibody specific... -
Choosing the B(right)est Fluorescent Protein: Photostability
TypeBlog PostPublishedJune 8, 2017, 2:30 p.m....This can differ between fluorescent proteins in the same spectral class (Cranfill et al, 2016). This non-...described by a mono-exponential decay (analogous to radioactive decay). Consequently, the intensity decay is ...comparison of several fluorescent proteins of the same spectral class on the microscopy system and in the... -
A Guide to Designing a Scientific Poster: Content, Layout, and More
TypeBlog PostPublishedOct. 22, 2019, 12:20 p.m....your poster. While a poster generally contains the same sections as a primary research article, it’s important...can be accomplished with an exciting title and attractive figures. Next, the poster should be legible from...four column style, and in all cases the idea is the same; the reader will navigate the poster from column... -
AAVs CREATed for Gene Delivery to the CNS and PNS
TypeBlog PostPublishedSept. 28, 2017, 2:01 p.m....under the control of an inducer, i.e. tet-off transactivator (tTA), is injected at a high dose to maximize...An AAV expressing the inducer (i.e. tet-off transactivator) is co-injected. The dose of this AAV can be... Central PMCID: 5088052. 3. Lentz TB, Gray SJ, Samulski RJ. Viral Vectors for Gene Delivery to the Central... -
Transferable Skills Guide: Identifying Your Transferable Skills
TypeBlog PostPublishedJan. 11, 2018, 3:29 p.m....paying bills and having a life outside my job. The same can be said about teamwork, communication, writing... every piece of work you touch, and be a more attractive candidate for that ideal job. How can you identify...process. You stepped up and got your team on the same page. This came naturally to you. How is this new...