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Guinea pig bone marrow sample with two megakaryocytes (arrows). Image courtesy of CDC PHIL ID#14465

New production method for lentivirus overcomes the barrier to using a promising pseudotype for transducing stem cells more effectively

What’s the problem, scientifically speaking? While hematopoietic stem and progenitor cells (HSPCs) have great potential for development into cell-based therapeutics, they are very difficult to transduce. This technical problem has slowed researchers’ ability to turn HSPCs into the next generation of medicines. What have been the technical barriers? The problem with transducing HSPCs has been […]

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Chemiluminescent Western Blot

Secrets to Chemiluminescent Western Blotting

What you need to know to optimize your chemiluminescent Western blots Chemiluminescence remains the most frequently used method to detect target proteins on Western blots. Many reagents are commercially available for chemiluminescent detection but all share basic characteristics. The secondary antibody is labeled with an enzyme, usually horseradish peroxidase (HRP). After incubation with the secondary […]

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effect of blocking agent

Getting Rid of the Noise: Western Blot Blocking

Why is the Western blot blocking step important? After a sample has been transferred from the gel to a blotting membrane, most Western blotting protocols include a blocking step before incubating the blot with the primary antibody. Understanding how blocking works is key to maximizing the quality of your Western blot, improving specificity, sensitivity, and […]

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Western Blot Normalization

Quantitative Westerns: What is the best way to normalize your Western blot?

Far from being an “is-it-there-or-not” technique, modern digital detection instruments can make Western blotting reproducible and quantitative. By working within the linear dynamic range of your detection method and normalizing the data to control for variations in protein load and membrane transfer, you can get truly quantitative results. But what is the best way to […]

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Western Blotting