Band Warping from Well Overloading (Western Blot (Misshapen / Uneven Bands))

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Individual lanes show warped or distorted bands while adjacent lanes appear normal. Affected lanes may show smearing, broadening, or lateral spreading of protein bands. Common causes: 1) Excessive protein amount loaded exceeding well capacity 2) Uneven protein loading across different wells 3) Sample volume exceeding well volume capacity 4) High protein concentration causing local gel matrix saturation Solutions: 1) Do not exceed recommended well loading capacity (typically 20-30 µg total protein for mini-gels) 2) Refer to gel manufacturer and comb size specifications for maximum sample volumes 3) Perform protein quantification (BCA or Bradford assay) to ensure equal loading 4) Dilute high-concentration samples to appropriate levels before loading