I analyzed IL-1b in whole cell lysate through western blot and treated LPS as a positive control.
Predicted size of IL-1b is 17/31 kD (mature/pro-form).
But in the result, band around 40 kD was detected.
So i think that band represents IL-1b but, because size of the band is different from predicted one, i cann't be sure whether that really shows IL-1b or not.
Is there any one who give me advice?
Hi Suyeol,
The predominant molecular form of IL-1beta found in cell lysates is the precursor (uncleaved) form that should be detected at approximately 35-37 kDa so very close to your finding. One good way to identify a band in WB is to treat half of your cell population with the relevant siRNA.
Best wishes,
Claire
I've always looked for a 34kD band for my IL1b precurser band, as shown in FIgure 1A in the paper linked to this answer. One thing you want to pay special attention to is that your LPS treatment is indeed upregulating the precurser expression of IL1b, as it should be little to none in non-treated samples.
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