...Lab plasmids Scot Wolfe Lab plasmids Detailed Information Protocols & Resources Contents of Kit Original...

...622-30. A benchmarked protein microarray-based platform for the identification of novel low-affinity extracellular...

...promoter, and so on. The following table provides information for the assembly standards most commonly used...

...specific DNA sequences. Ligand binding triggers a conformational change in the receptor and results in activation...

...Tagging in Human Cells Find additional cell line information from the Allen Institute for Cell Science data...

...search our full site. Find resources for more information on shRNA design and delivery. Constitutive Lentiviral...

... Please see the following reference for more information: A Human Platelet Receptor Protein Microarray...

...arrays in a 2012 issue of Nature Protocols . More information and help can be found at TALeffector Resources...

...specimens. These technologies have the potential to transform neuroscience research by bringing the precision...

...Promoter Plasmids using the table below. Additional information is available for each construct and can be accessed...

...added to this list? Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve this... tags such as fluorescent protein (FP) tags. Information on genes and associated neurodegenerative diseases...SOD1-VC SOD1 Venus CMV ALS Tiago Outeiro 89780 isoform PKC Gamma (JCE692) PRKCG PKC1 Spinocerebellar ataxia...

... the following spreadsheet, fill out as much information as possible on the sequences, and email the file...

... is stable at 4°C for up to one year. The powder form of polybrene is stable at 4°C for several years....provided for your convenience. See “warranty information” in appendix. Table of Contents A. pLKO.1-TRC...Digesting pLKO.1 TRC-Cloning Vector C.4 Ligating and transforming into bacteria D. Screening for Inserts D.1 Recommended... TRC-Cloning Vector I.2 Recipes I.3 Warranty information Back to Top A. pLKO.1-TRC Cloning Vector A.1 ...pCMV-VSVG from Robert Weinberg’s lab. For more information, visit Addgene’s Mammalian RNAi Tools page. Several...Measure the DNA concentration. C.4 Ligating and Transforming into Bacteria Use your ligation method of choice...final volume Incubate at 16°C for 4-20 hours. Transform 2 μL of ligation mix into 25 μL competent DH5 ...

...light chains. These four domains come together to form a “Y” shape, as shown in Figure 1. The two arms ...the five human isotypes. Some isotypes naturally form multi-antibody units, while others exist as monomers...may want to identify or collect the protein in any form it may be present in. However, polyclonals can vary...antibody, for each antibody used. Blank controls in the form of sample buffer must also be run. Unique samples...fluorophores. Horseradish peroxidase (HRP) HRP performs a chemiluminescent reaction in the presence of...activated and its output measured. This method can be performed using the indirect detection method via a conjugated... able to recognize the protein in its native conformation. Attachment of the antibody to the beads is ...

...hort P alindromic R epeat (CRISPR) systems, which form an adaptive immune system in bacteria, have been...genome engineering. Find essential background information on CRISPR and the basics for planning your first...

...from Arabidopsis thaliana is fused to truncated forms of cytosolic STIM1 to control calcium channels ~...UVR8 domains UVR8 plant photoreceptor protein that forms photolabile homodimers with slow reversal kinetics...the common actuators used in optogenetics. For information on sensors, check out our Biosensors Plasmid ...blue-shifted channelrhodopsin from Platymonas subcordiformis 445 CoChR Channelrhodopsin from Chloromonas...465 SwiChRca Chloride-conducting channel, Step-waveform inhibitory ChR with mutation C128A (derived from...widely used Cry2-CIB1 system, light induces a conformational change in cryptochrome Cry2 to permit CIB1 ...cryptochromes with light-induced changes in protein conformation/dimerization. In addition to the applications...

... Day 1: Transform, recover, set up overnight growth (Estimated time 2-3 hours) Transformation should be...of 5 mL (3 mL SOC + 2 mL transformed Endura from two separate transformations). Pro-Tip Do not pipette...Follow this protocol to perform amplification of CRISPR pooled plasmid libraries in Escherichia coli ...assumes familiarity with standard bacterial transformation and basic knowledge of the library being amplified...amplified. Please see our protocol on Bacterial Transformation . The generalized pooled library amplification... be performed at the end of the day to ensure that growth times are limited. Day 2: Harvest cells and ...using a new vented falcon tube for every two transformations. Each of the four 14 mL Vented Falcon Tubes...

... of cold 4% paraformaldehyde in PBS on ice for 15 min . Pro-Tip While 4% Paraformaldehyde fixation works...rocking platform. Permeabilize cells for 10 min at room temperature ( RT ) on a rocking platform in 500...rocking platform. Section 3: Labeling with antibody Block for 20 min at RT on a rocking platform in 500...labeling HeLa cells for a target protein using the formaldehyde fixation method. The protocol may need to be...specific products or equipment. Inclusion of this information is solely for transparency intended to support...Pipette controller Pipette tips and pipettes Rocking platform Tweezers Fluorescent microscope 0.5–10 µL single...coated coverslips HeLa cells 24-well plate 4% Paraformaldehyde 5 mg/mL 4′,6-diamidino-2-phenylindole (DAPI...

...incubation, transform DH5α E. coli cells by heatshock with the plasmid of interest. See our transformation page...for a detailed heatshock transformation protocol. Plate 50 µL of transformed E. coli /rescue media suspension...also like... Making LB Agar Plates Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture...plating antibiotic over-agar for the selection of transformed E. coli . Over-agar spreading of antibiotic makes...investigator to conveniently plate and select transformed cells containing plasmids differing in their ... sterile water (or other antibiotic) E. coli transformed with a plasmid containing the carbenicillin (...for 18 hours. Day 2 Observe plates for colony formation. Shown below are the results from an experiment...

... a shaking platform. Wash the membrane 3x for 5 min in 1X TBST at RT on a shaking platform. Section 6:... a shaking platform. Wash the membrane 3x for 5 min in 1X TBST at RT on a shaking platform. Prepare the...specific products or equipment. Inclusion of this information is solely for transparency intended to support...supply iBlot 2 Gel Transfer Device Roller Spatula Platform shaker Cold room Gel imager -80 °C freezer Reagents...the manufacturer's instructions for additional information specific to your antibody, such as ideal blocking...between antibodies. Refer to the manufacturer's information before running your experiment. Remove the membrane...overnight in primary antibody at 4 °C on a rocking platform. Pro-Tip Primary antibody incubation can be reduced...