Rinehart Lab Phosphoprotein Reagents
Type
Collection
...laboratories performing phosphoproteomic experiments in need of reference spectra or phosphopeptide standards...
We narrowed to 829 results for: EED
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Cre-Lox and Other Site-Specific Recombinases
TypeCollection...the stop codon is excised, and gene expression proceeds. Gene Knockout: Placing the loxP sites on either... -
Optogenetics AAV Preps
TypeCollection...plasmids and we’ll make the viral vector prep you need. Or fill out our form to let us know what you would... -
Retrograde AAV viral preps
TypeCollection...plasmids and we’ll make the viral vector prep you need. Or fill out our form to let us know what you would... -
Plasmids for Stem Cell Research
TypeCollection...Pluripotent Stem Cells From Adult Blood Cells Under Feeder-Free and Xeno-Free Culture Conditions: A Clinically... -
Biosensor AAV Preps
TypeCollection...plasmids and we’ll make the viral vector prep you need. Or fill out our form to let us know what you would... -
Fluorescent Protein Guide: Empty Backbones
TypeCollection...label one fusion protein with different colors as needed for your experiments. Learn more about the flexibility... -
Plan Your Experiment
TypeGuide...population need to be identical (clonal) or is a mixed population okay? Does the edit need to be homozygous...interference Needs good primer design Western blot Knockouts Activation/interference Needs a good antibody...other organisms. As with most experiments, you first need to have a biological question in mind. Then you ...will help you narrow down which components you’ll need for a given experiment. In the table below, we highlight...dCas9 to be fused to transcriptional activators that need to be within a given range of the transcription ...HEK293) cells. For more difficult cell types, you may need to look into other options. Browse Addgene's collection...cells are not transcribing DNA to mRNA, nor do they need to translate mRNA to protein. Instead, you directly... -
Antibody Guide
TypeGuide...process may need to be optimized for individual assays. Clear cell debris by sedimentation. Proceed with IP..., an antimicrobial agent, can block some amines needed for conjugation reactions and should be removed...chosen. Signal Amplification The amount of signal needed from a specific antibody can vary depending on ...category, including an outline of the basic steps needed for each application, important controls, and any...samples to make proteins available. Tissue samples may need additional processing steps, such as sonication....signal levels. In ChIPseq, an input control will need to be collected before immunoprecipitation. Antibody...from. Then consider the following questions: Do you need polyclonal or monoclonal antibodies? Will you be... -
Adenovirus Guide
TypeGuide...therefore needs to be supplied through other means. In order to produce rAdV vectors, E1 needs to be expressed...especially when needing transgenes to express for a shorter period of time or when needing to package larger...starts and primarily encode for structural proteins needed to assemble icosahedral capsids and build new virions...acronyms, so if you're new to viral vectors or simply need a refresher, we've included a glossary at the end... In order to produce rAdV vectors, you generally need two plasmids: Shuttle plasmid (also known as transfer...produced in high titers for their use in experiments. Needle sticks and ripped gloves are common points of entry...system for generating recombinant adenoviruses . Proceedings of the National Academy of Sciences of the United... -
Gamma-Retroviral Vector Guide
TypeGuide...acronyms, so if you're new to viral vectors or simply need a refresher, we've included a glossary at the end...In order to produce gamma-retroviral vectors, you need three plasmids (Figure 2): Transfer plasmid — contains...stably express gag-pol and/or env , eliminating the need to deliver these genes on separate plasmids ( in...with contact to mucous membranes or broken skin. Needle sticks and ripped gloves are common points of entry...clinical-grade gamma-retroviral vector purification by high-speed centrifugation for CAR T cell manufacturing. Molecular...specificity of infection for CD4-expressing cells. Proceedings of the National Academy of Sciences , 94 (16)...the beginning of R, has a 5' cap structure, and proceeds through U5 and the rest of the provirus. Some ... -
CRISPR Guide
TypeGuide...requirements for a CRISPR knockout assay are simple. You’ll need a Cas enzyme and a gRNA specific to the gene of ...Cas9-gRNA complex binds a putative DNA target, the seed sequence (8–10 bases at the 3′ end of the gRNA targeting...) will begin to anneal to the target DNA. If the seed and target DNA sequences match, the gRNA will continue...Mismatches between the target sequence in the 3′ seed sequence inhibit target cleavage, while mismatches...NHEJ directly ligates the break ends without the need for a homologous template. Both NHEJ and HDR have... the depositing lab. After amplification, you’ll need to use next-generation sequencing (NGS) to verify...matches that of the original library. Next, you’ll need to generate lentivirus containing the entire library... -
Lentiviral Vector Guide
TypeGuide...acronyms, so if you're new to viral vectors or simply need a refresher, we've included a glossary at the end...table . In order to produce lentiviral vectors, you need three (or four, for third generation) plasmids: ...heterologous promoter (often CMV or RSV), eliminating the need for transactivation by Tat. Most third-generation...Biology Reference . Production After cloning, you will need to produce the lentiviral vectors from your plasmids...with contact to mucous membranes or broken skin. Needle sticks and ripped gloves are common points of entry...definition, at the beginning of R, is capped, and proceeds through U5 and the rest of the provirus. Third-generation... -
Adeno-associated virus (AAV) Guide
TypeGuide...acronyms, so if you're new to viral vectors or simply need a refresher, we've included a glossary at the end...order to produce rAAV vectors, three plasmids are needed: Transfer plasmid (also known as cis plasmid) —.... The cell line used for rAAV vector production needs to express the adenovirus E1 gene, which is crucial...especially if any additional regulatory elements are needed. One strategy to overcome this obstacle, has been...with contact to mucous membranes or broken skin. Needle sticks and ripped gloves are common points of entry... -
Molecular Cloning Techniques
TypeGuide...is usually an ideal approach. This may be due to speed, cost, availability of starting materials, or just...compatible ends together. TOPO cloning thus does not need restriction enzymes or an exogenous ligase, providing... of PCR products into donor plasmids without the need for restriction enzyme cloning. Read more in our...the chosen orientation at a time, and without the need for any unwanted sequence at the junctions (such... -
Addgene's eBook Collection
TypeGuide...Not sure where to start? Need a quick reference? Addgene's eBooks are here to help!... Collection Not sure where to start? Need a quick reference? Addgene's eBooks are here to help! 0 of 0...101 blog series, this eBook covers everything you need to know about plasmids, from parts, nomenclature... -
Chemogenetics Guide
TypeGuide...back-metabolism of CNO to clozapine, demonstrating the need for alternative DREADD ligands. Compound 21, deschloroclozapine...while PSAMs have more than one domain, both of which need to be expressed. However, neuronal control through...experiment, an AAV-encoded chemogenetic plasmid may need to be targeted to specific tissues, cell types or... -
Optogenetics Guide
TypeGuide...Channelrhodopsin from Volvox carteri 570 Chronos High-speed, light-sensitive channel from Stigeoclonium helveticum...When designing your optogenetics experiment, you’ll need to pick both an opsin and a delivery system. Here...experimental design will determine whether you’ll need short or long periods of neuronal activation/inactivation... -
Antibodies 101: Flow Compensation
TypeBlog Post...will need compensation? To know if you will need to compensate in your flow panel, you will need to know...would say that APC is “bleeding,” or spilling over, into APC-Cy7 channel and needs to be compensated out...this bleed. A compensation value of 3.7 for APC into APC-Cy7 applied next, which fixed the bleed in this...fluorescent markers are read by the same laser, they will need to be compensated for one another. This overlap ...fluorophores FITC and PE on the same laser, you will need to compensate between the two of them because they...reads them out on different lasers, you should not need to compensate between them. Another rule of thumb...of the fluorescent markers are similar, you will need to compensate between them (although there are exceptions... -
How to Negotiate a Successful Lab Start Up Budget
TypeBlog Post...Identify the skills needed to use these tools Outline the knowledge and skill sets needed to do your science...that you’ll need to tap for your future research team. For example, if you’re going to need a tech because...Experiment Designs Consulting, Inc. “How much do you need [for your lab start up]?” A hiring chair or dean...doesn’t have to be this way, if you know what you need. Most often, early career scientists aren’t trained...instead have our clients focus on their experimental needs and consider a budget as a resource checklist. A... budget is an account of the material resources needed to accomplish your science, and can include: equipment...discussion first defines the material resources needed, then determines how much those resources will ...