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Protocol for Quantitative Western Blots

Strategy

Choose a Normalization Strategy

Prepare Samples

Prepare Samples

  • Tip: Determine protein concentration with an assay.
Perform Electrophoresis

Perform Electrophoresis

Transfer to Membrane

Transfer to Membrane

Incubate with Primary Antibodies

Incubate with Primary Antibodies

Wash

Wash

  • Tip: Clean Incubation boxes with methanol before you use them to prevent contamination.
Incubate with Secondary Antibodies

Incubate with Near-Infrared Fluorescent Secondary Antibodies

Wash

Wash

  • Tip: Use freshly-made buffers to prevent contamination.
Detect with Odyssey® imaging system

Detect with Odyssey® imaging system

Analyze with Image Studio™ Software

Analyze with Image Studio™ Software

Store or Archive Blots with Western Blot Storage Bags

Store or Archive Blots

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