Tech Tips Video: Milk or BSA? Choosing a blocking protein for Western Blot


Posted by Ken B on Sep 5, 2018 3:00:00 AM

So you've set the timer to five minutes for the first of three TBST washes for your western blot membrane. Now what? Sure, you could check your email or social media for the 30th time before lunch. Or you could do something informative, like check out a CST Tech Tips video! This is a new short video series featuring the same scientists who develop and validate CST antibodies, here to offer insights and protocol tips.

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Topics: Protocols, Western Blot, techniques, Tech Tips

Are you ready to transfer your protein? Wet vs. semi-dry transfer


Posted by Neha G on Apr 18, 2018 3:15:00 AM

 You are all amped up to run a western blot to identify “your favorite protein.” The lysates have been run and proteins separated by SDS-PAGE. Now it’s time to transfer proteins from the gel to the membrane, and you’re sitting wondering….wet or semi-dry?? Or maybe you are better prepared than I was as a graduate student and you already know your next step, in which case you are aware of the pros and cons of wet and semi-dry transfer.

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Topics: Science Education, techniques, Western Blot, Protocols

Have You Ever Wondered...How Do I Choose a Loading Control?


Posted by Chris S on Mar 21, 2018 6:15:00 AM

For those who wish to bring a measure of rationality to the western blot data in the literature, loading controls can induce a fair amount of hand wringing. There are some obvious criteria. First, the protein level of the loading control must not be altered by experimental conditions. Second, the molecular weight of the control must be different from the protein of interest. Third, and most importantly, the signal detected for both the loading control and the protein of interest must be in a linear range, otherwise you’ll find yourself with burnt-out bands that you’ll be unable to quantify. 

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Topics: Western Blot

Have you ever wondered: Am I setting up my loading controls correctly?


Posted by Ken B on Nov 1, 2017 3:00:00 AM

You’re gathering data from all your experiments and preparing to present to your advisor and thesis committee at your annual progress report. You have an interesting hypothesis, and you have a validated antibody that recognizes your target protein on a western blot (WB). The molecular weight of the band is correct, and the expression of the target protein changes just the way you predicted it would. Now, you know — and you’d bet the house on it — when that powerpoint slide comes up, someone on your committee is going to ask about loading controls.

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Topics: Western Blot, Antibody Performance, Companion Reagents, Protocols, techniques

Have You Ever Wondered: What's The Lowdown on Phospho-Specific Antibodies?


Posted by Ken B on Jul 26, 2017 3:20:00 AM

Early exploration of unmapped biological signaling pathways were carried out using radiolabeled phospho-imaging. The development of phospho-specific antibodies to detect and quantify protein phosphorylation made life easier for researchers (less 32P waste to deal with), but the interpretation of data from these experiments comes with its own set of caveats. 

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Topics: Western Blot, Antibody Performance, Post Translational Modification, techniques, Primary Antibodies

The Cost of a Failed Western Blot


Posted by Claire S on Sep 14, 2016 3:00:00 AM


Thinking about lab expenses isn’t as enjoyable as investigating your favorite signaling pathway. However, because research money is hard to come by, it is something that should be considered when picking your reagents for western blotting. It makes sense to keep the quality of your primary antibody in mind, because the success of the entire experiment depends on the antibody being reliable, specific, and sensitive. If the antibody does not perform as expected your experiment may fail . . . and the cost of a failed western blot may be more than you think.

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Topics: Antibody Performance, Antibody Validation, Western Blot, techniques, Reproducibility

The Cost of a Failed Western Blot


Posted by Claire S on Aug 13, 2015 8:13:02 PM


Thinking about lab expenses isn’t as enjoyable as investigating your favorite signaling pathway. However, because research money is hard to come by, it is something that should be considered when picking your reagents for western blotting. It makes sense to keep the quality of your primary antibody in mind, because the success of the entire experiment depends on the antibody being reliable, specific, and sensitive. If the antibody does not perform as expected your experiment may fail . . . and the cost of a failed western blot may be more than you think.

READ MORE >

Topics: Antibody Performance, Antibody Validation, Western Blot, techniques

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