...using ready-to-use bio-safe Coomassie G-250 stain (Addgene uses SimplyBlue SafeStain) and ImageJ software....using ImageJ software and a standard curve is generated. The ratio of the antibody protein content to ...reagents, and methods that we use in our lab at Addgene. Equipment and reagents from other vendors should...accommodate slight differences between products. Addgene does not endorse or recommend specific products... may not be functional. This sample would fail Addgene QC and would not be used. Section 3: Image Analysis...11469.296 4655.113 16642.522 16124.409 96.887% Pro-Tip Addgene only uses recombinant antibody preps that have ...

...reagents, and methods that we use in our lab at Addgene. Equipment and reagents from other vendors should...accommodate slight differences between products. Addgene does not endorse or recommend specific products...intended to support reproducibility in science. General Considerations The ELISA can be completed in a ...The color will change from yellow to blue and generally takes between 15 to 30 min. High concentrations...spectrophotometer. Use the spectrophotometer to generate a standard curve . If you don’t see any color ...

...reagents, and methods that we use in our lab at Addgene. Equipment and reagents from other vendors should...accommodate slight differences between products. Addgene does not endorse or recommend specific products... optimal antibody concentration will vary but generally ranges from 1-10 µg/mL. Add 500 µL of the diluted... optimal antibody concentration will vary but generally ranges from 1-10 µg/mL. Add 500 µL fluorescently-labeled...

...reagents, and methods that we use in our lab at Addgene. Equipment and reagents from other vendors should...accommodate slight differences between products. Addgene does not endorse or recommend specific products...intended to support reproducibility in science. General Guidelines Acrylamide percentage varies in SDS-...depending on the size of your cell pellet but will generally be between 250–1000 µL . The ideal lysis buffer...

...designed by the Addgene nonprofit plasmid repository and is being shared outside of Addgene for informational...administrators, or anyone else associated with Addgene, can be held responsible for your use of the information...

...achieves this by using centrifugal force, which is generated by spinning the sample at a high speed. Being ...specific temperatures. This protocol will cover the general procedure and features to keep in mind when using... larger containers. Ultracentrifuges are also generally floor sized centrifuges that can hold larger containers...

...designed by the Addgene nonprofit plasmid repository and is being shared outside of Addgene for informational...administrators, or anyone else associated with Addgene, can be held responsible for your use of the information...

...Use this protocol to generate lentivirus...downstream applications such as stable-cell line generation. Workflow Timeline Day 0: Seed 293T packaging...adherent cells) 0.45 μm polyethersulfone filter, Nalgene, 565-0010 Microcentrifuge tubes, Neptune 3745.X...

...reagents, and methods that we use in our lab at Addgene. Equipment and reagents from other vendors should...accommodate slight differences between products. Addgene does not endorse or recommend specific products...30 kDa MWCO buffer exchange column. (Optional) Regenerate the column by washing with 25 mL of Protein A...

...your DNA sample concentration and purity using Addgene’s DNA Quantification Protocol....ratio of nucleic acid to protein (260/280) is generally used as an indicator of the purity of DNA samples...

...annealed can be cloned directly into the overhangs generated by restriction digest of existing sites in the... additional bases to complement the overhangs generated when digesting the vector with EcoRI and SalI ...

...downstream applications. Safety Warnings Lentivirus is generally considered biosafety level 2+. Please ensure that...adherent cells) 0.45 μm polyethersulfone filter, Nalgene, 565-0010 (for viral preps, if prep was not previously...

...Recombinant antibodies are created by cloning antibody genes from specific hybridoma cells into plasmids. Plasmids... tubulin (sometimes referred to as housekeeping genes). This allows for relative comparison of expression...Science Guides Antibody Guide Antibody Guide View Addgene's Antibody Collection Jump To: Antibody Guide Production...with myeloma cells (immortalized B cells). The genetic reorganization that creates variety in the antibody... be produced from plasmids, with an antibody’s genetic code inserted into the plasmid. This produces the...antibody is the antigen binding site. In B cells, the genetic region responsible for the variable region goes...epitopes on one antigen. Monoclonal antibodies are generated from cultured B cells, where each culture should...

...consideration the information above, primers should generally have the following properties: Length of 18-24...

... medium. This dot is composed of millions of genetically identical bacteria that arose from a single bacterium...

...temperature between 4°C and room temperature. This general protocol suits almost any water bath. Video Watch...

...your plasmid. Many DNA analysis tools, including Addgene’s Sequence Analyzer , allow you to identify which...

...cut), use a sequence analysis program such as Addgene's Sequence Analyzer . Determine an appropriate reaction...

..., BD BD300912 0.45 μm polyethersulfone filter, Nalgene, 565-0010 (for viral preps, if prep was not previously...

...consider using a multichannel pipette. Please watch Addgene’s protocol on multichannel pipetting to learn more...