HSP70 | positive control/quantitation standard
AS08 371S | protein standard for quantitation and positive control
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Standard curve: 3 loads are recommended (eg 0.1, 0.2, 0.3 pmol). Adjust range to fit your samples and your experiment.
Positive control: a 2 μl load per well is optimal for most chemiluminescent detection systems. Higher standard concentration needs to be used to allow detection by Coomasie stains. Such gels with higher standard concentration can not be used for quantitation using chemiluminescence.
This standard is stabilized and ready and does not require heating before loading on the gel.
|Expected | apparent MW||
|Not reactive in|
Concentration: after re-constitution with sterile milliQ water final concentration of the standard is 0.15 pmoles/µl
|Selected references||to be added when available, product released in September 2015|
Quantitation: When quantitated standards are included on the blot, the samples can be quantitated using the available software. Excellent quantitation can be obtained with images captured on the Bio-Rad Fluor-S-Max or equivalent instrument using Bio-Rad QuantityOne software. The contour tool is used to the area for quantitation and the values are background subtracted to give an adjusted volume in counts for each standard and sample. Using above protocol linear standard curves are generated over 1-1.5 orders of magnitude range in target load. It is important to note that immunodetections usually show a strongly sigmoidal signal to load response curve, with a region of trace detection of low loads, a pseudolinear range and a region of saturated response with high loads. For immunoquantitation it is critical that the target proteins in the samples and the standard curve fall within the pseudolinear range. Our total detection range using this protocol spans over 2 orders of magnitude, but the quantifiable range is narrower.
Quantitative western blot: detailed method description.
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