...science. General Guidelines Acrylamide percentage varies in SDS-PAGE gels. Use lower percentage acrylamide...buffer. Pro-Tips The volume of lysis buffer will vary depending on the size of your cell pellet but will...between 250–1000 µL . The ideal lysis buffer will vary depending on the cellular location of the protein...sample. Pro-Tip The ideal total protein loaded will vary between samples and target proteins but is typically... 1X TBST. Pro-Tip The ideal blocking buffer will vary between antibodies. Refer to the manufacturer's ...blocking buffer. Pro-Tip The optimal concentration will vary between antibodies but is typically between 1–10...

...increases reproducibility, as it eliminates the variation associated with repeated transient transfection...lots can promote or inhibit transfection. Test a variety of brands and FBS lot numbers to find one suitable...transduction efficiency. Infection efficiency will vary between cell lines. It is not recommended that lentiviral...that already contain 0.5 mL of virus solutions at various dilution. Make sure to use the polybrene-containing...transgene has integrated in different locations in the various cells in the culture. This is because lentiviral...of the lentivirus used, cells may also exhibit varying numbers of integration events. In other words, ... transgene in their genome. This can result in varying expression levels of the transgene from different...

...will vary depending on the sample type and the protein of interest. You may need to try a variety of fixation... Pro-Tip The optimal antibody concentration will vary but generally ranges from 1-10 µg/mL. Add 500 µL... target. The optimal antibody concentration will vary between antibodies. Review the manufacturer's instructions...

...reagents. Once produced, lentivirus can be used for a variety of downstream applications such as stable-cell ...FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to find one suitable ... cm dish. Pro-Tip There can be batch to batch variation when making the PEI working stock, therefore the...transfect cells with a fluorescent plasmid using a variety of ratios. Check the cells 1-2 days after transfection...

...Molecules absorb different wavelengths of light to varying degrees and most have a specific wavelength that...spectrophotometer between samples if they are expected to vary significantly in concentration. DNA dissolved in...in water is going to have a greater variance in concentration readings than a DNA sample dissolved in ...

...developed for 293T cells but can be adapted to a variety of target cell lines and selection markers. Note...FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to find one suitable ...viability and transduction efficiency. Titer will vary between cell lines. It is not recommended to subject...

... enzymes, allowing scientists to target a wide variety of recognition sequences. For a list of many commonly... 1 µg of DNA. The total reaction volume usually varies from 10-50 µL depending on application and is largely...is due to "Star Activity" and can happen for a variety of reasons, including high glycerol concentration...

...FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to find one suitable ...new batch of 1 mg/mL PEI prepared. There may be variation between batches of PEI depending on the user, ...transfect cells with a fluorescent plasmid using a variety of ratios. Check the cells 1–2 days after transfection...

...developed for A549 cells but can be adapted to a variety of target cell lines and selection markers. Note...FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to find one suitable ...viability and transduction efficiency. Titer will vary between cell lines. It is not recommended that lentiviral...

...approximately 1:3. Since the number of base pairs for each varies, it is difficult to calculate this based on DNA...for each plasmid you are trying to create, with varying ratios of recipient plasmid to insert. It is also... plating on the appropriate antibiotic and try varying the amount of recipient plasmid to insert in the...

...fluctuations and to minimize well-to-well variability that result from a variety of causes, such as pipetting error...multichannel pipet to minimize pipetting error and variability Mix samples very well by pipetting back and forth...

...P8833 Target cells Varies based on your experiment Culture media for target cells Varies based on your experiment...to 20 μl serum-free OPTI-MEM TIP: You may want to vary the ratio of shRNA plasmid, packaging plasmid, and...experiment Materials for assay Varies based on your experiment Detailed protocols for preparing polybrene...

...approximately 1:3. Since the number of base pairs for each varies, it is difficult to calculate this based on DNA...for each plasmid you are trying to create, with varying ratios of recipient plasmid to insert. It is also...your transformation worked. You could also try varying the amount of recipient plasmid to insert. Isolate...

...optimal concentration of primary antibody to use will vary and needs to be empirically determined. If possible...buffer. Pro-Tip The ideal antibody concentration will vary and needs to be empirically determined. We suggest...concentrations. The ideal range of the standard curve will vary between targets and needs to be empirically determined...

...results in a heterogenous polyclonal population that varies in the number of integration events and the site...culture has, in theory, been transduced. There are various schools of thought on using selective medium throughout...monoclonal lines generated using this protocol will vary based on several factors, including the growth properties...

...vector DNA and insert DNA used in the ligation will vary depending on the size of each and their concentration...failed ligations. The following table indicates the various controls: Control Ligase Interpretation Uncut vector...

...recombinant antibody can be purified for use in a variety of applications. Sharing speeds science. We believe...to mix. Pro-Tip The optimal ratio of DNA:PEI may vary significantly and should be empirically determined...

...FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to find one suitable ...protocol. Sample Data Figure 1: HEK293T cells at various confluencies. This AAV production protocol should...

...experiment would need to be repeated to account for variability....

...in your protocol. How to set the temperature will vary depending on the model of water bath, but will likely...