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ELISPOT severe

Physical Membrane Damage and Artifacts

Symptom
Membrane shows physical damage, scratches, or irregular patterns. Spots may appear distorted or artifacts present as non-specific signals.
Common Causes
  1. 1 Flow rate on automated washer too high or manual pipetting too harsh
  2. 2 Pipette tips touching the membrane during washing or reagent addition steps
  3. 3 Membrane dried out at various experimental stages
  4. 4 Membrane not pretreated with 35% ethanol before coating
Solutions
  1. 1 Use gentler washing procedure; reduce flow rate on automated washers
  2. 2 Exercise care during all pipetting steps, especially automated washing
  3. 3 Keep membrane hydrated at all stages; never allow to dry out
  4. 4 Pretreat membrane with 35% ethanol followed by 3× PBS washes
Related Video (2)
Mabtech ★ 85
ELISpot tutorial: step-by-step full assay protocol
"Step-by-step protocol tutorial with handling tips directly addresses proper technique to avoid physical membrane damage from incorrect pipetting and washing procedures"
Animated biology With arpan ★ 72
Elispot assay ( assay to determine cytokine producing cells)
"Overview of ELISpot assay principles and basic procedure provides foundational understanding of membrane handling requirements and potential artifact sources"
Source: abcam.com ↗
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