How to Dilute WB Samples for Equal Loading When Initial Amounts Vary by Gray Value

To standardize the sample loading amount in WB based on gray value and ensure equal protein loading, follow these steps:

 

1. Determine the target gray value

Select a target gray value, typically the lowest among the samples.

 

2. Calculate the dilution ratio

For each sample, determine the dilution ratio by calculating the ratio of its gray value to the target gray value using the following formula:

Dilution ratio = Sample gray value / Target gray value.

 

3. Perform dilution

Adjust each sample by diluting it with an appropriate buffer solution according to the calculated dilution ratio.

 

Following this procedure, the sample loading amount will be normalized based on its initial gray value, leading to consistent signal intensity across all samples during electrophoresis. If further clarification is required, please feel free to seek additional guidance.

 

MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.

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