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We narrowed to 347 results for: SAM activators

Showing: 301 - 320 of 347 results
  1. Antibodies 101: Polyclonal Antibodies

    Type
    Blog Post
    ...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 ...
  2. Antibodies 101: Chimeric Antibodies

    Type
    Blog Post
    ... 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....
  3. Lentiviral Vector Guide

    Type
    Guide
    ... and actively dividing cell types, whereas gamma-retroviruses can only infect mitotically active cell ...packaging plasmids. tat in trans Trans-activator; binds TAR to activate transcription from the LTR promoter...the transfer plasmid, and this requires Tat for activation. Second-generation plasmids include (Figure 3...lentiviruses most often integrate into transcriptionally active regions, regions recently involved in translocation...Instead of selection by an antibiotic, fluorescence-activated cell sorting (FACS) is used to sort cells expressing...CRISPR libraries, including knockout, inhibition/activation, and prime and base editing. Browse Addgene's... systems do not require the HIV protein Tat to activate the promoter on the transfer plasmid. Many of ...
  4. Protocol - How to Design Primers

    Type
    Protocol
    ...must selectively block and unblock repeatedly the reactive groups on a nucleotide when adding a nucleotide...primer is used to amplify a eukaryotic genomic DNA sample. However, a primer should not be too long (> 30...
  5. Gamma-Retroviral Vector Guide

    Type
    Guide
    ... and actively dividing cell types, whereas gamma-retroviruses can only infect mitotically active cell ...are found in proto-oncogenes. This can lead to activation of the oncogene and possible development of cancer...Instead of selection by an antibiotic, fluorescence-activated cell sorting (FACS) is used to sort cells expressing...site selection is driven by strong enhancers and active promoters. Nucleic Acids Research , 42 (7), 4257... (unique 3’) contains sequences necessary for activation of viral genomic RNA transcription. R is the ...cis Unique 3’; contains sequences necessary for activation of viral genomic RNA transcription. Removal of...Both lentiviruses and gamma-retroviruses use the same packaging genes. However, they are different viruses...
  6. Corralling and Cornering the Goldilocks Antibody

    Type
    Blog Post
    ...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 ...
  7. Antibody Guide

    Type
    Guide
    ...fluorophores are available in a range of colors activated by different wavelengths, allowing for easy differentiation...conjugated with HRP. Wash and block the membrane. Activate the HRP with a substrate kit. Image the membrane...antibody, followed by incubation and wash steps. Activate the conjugate and read the output via a plate ... antigen and the conjugate, typically HRP, is activated and its output measured. This method can be performed...secondary antibody is added. The conjugate is activated and the output is read. A standard curve with ...deposited into tubes or plate wells) by fluorescence activated cell sorting (FACS). Figure 7: Flow cytometry ...application Collect and lyse samples to make proteins available. Tissue samples may need additional processing...
  8. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    ...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...
  9. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    ...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...
  10. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...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...
  11. Antibodies 101: Validation

    Type
    Blog Post
    ...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 ...
  12. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    ...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 ....
  13. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...gene activity. Latent infection A state where a pathogen is present in an organism but not actively causing...is crucial for efficient AAV replication as it activates the necessary cellular machinery to produce large...non-dividing cells while reducing the risk of immune activation, both key advantages of rAAV vectors. AAV Types...Optogenetics Biological technique to control the activity of cells, often neurons, with light. Pseudotyping... new window) McCarty, D. M., Monahan, P. E., & Samulski, R. J. (2001). Self-complementary recombinant ...Link opens in a new window) Wu, Z., Asokan, A., & Samulski, R. J. (2006). Adeno-associated virus serotypes...in cis refers to genetic elements located in the same plasmid as the gene of interest. in trans In the...
  14. Cultivating Community Science at BosLab

    Type
    Blog Post
    ... 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 ...
  15. Molecular Cloning Techniques

    Type
    Guide
    ...exonuclease activity of T4 DNA polymerase. In LIC, the T4 DNA polymerase’s exonuclease activity creates “...free nucleotide. T4 will resume its polymerase activity, add back the free base, and become stuck at this...side of it into any plasmid that has the same sites in the same orientation within its MCS. Due to their...
  16. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ...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 ...
  17. Inside Addgene's 10 Year Anniversary Gala

    Type
    Blog Post
    ... 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...
  18. In Living Color: The Skinny on In Vivo Imaging Tools

    Type
    Blog Post
    ...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...
  19. Plasmids 101: Antibiotic Resistance Genes

    Type
    Blog Post
    ... 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...
  20. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    ...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...
Showing: 301 - 320 of 347 results