ProteinSimple eBook - 9

Simplifying Protein Analysis * Westerns and Total Protein Analysis

Big proteins are a BIG challenge with a traditional
Western blot. If you're spending too much time
troubleshooting the transfer of a high molecular
weight protein, you're not alone! The process of
optimizing gels and buffer systems and ﬁguring out
whether to use a tank or semi-dry transfer is a real
pain. And judging by the number of online forums
dealing with this topic, it's clear Western blotting of
high molecular weight proteins is an obstacle in the
road to publication-worthy data for many.
Our Simple Western assay for high molecular
weight proteins leaves no big protein behind!
Wes, Peggy Sue and Sally Sue have extended their
molecular weight range and can now detect
Figure 5. Panel of large proteins run with the
66-440 kDa pre-filled plate on Wes. We used
0.2 mg/mL of A10 lysate to detect dystrophin,
2 μg/μL of A431 lysate to detect BRCA2, 1.0 μg/μL
of A431 lysate to detect TRRAP, 0.25 μg/μL of K562
lysate to detect CBP, and 1.0 μg/μL of K562 to detect
DNA-PK and UBR5. All samples were denatured at
95 °C for 5 minutes. Analysis on Wes was complete
in under three hours.
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| GENengnews.com

proteins up to 440 kDa. And they do it with
the same simplicity, sensitivity, and reproducibility
they're known for (Figure 5).
We promised it was going to be simple, right?
Whichever instrument or assay you choose, you'll
get the separation you need, identiﬁcation of your
target protein, and truly quantitative data that
enables you to make accurate experimental decisions.
With 21CFR11 option, and a choice of throughput
Simple Western is the platform of choice of many
major pharma, CROs and academics laboratories.
Go ahead, move that mountain of gel apparatuses,
transfer tanks, and shakers off your bench and
Simple your Western. It's time. n
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