MECOM Western Blot Experimental Design Guide

Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before running
Expected band 138.1 kDa
Observed band Not reported — verify product WB image
Gel 8-10%
Positive control ⓘ Adipose tissue
Negative control ⓘ Target knockdown/knockout
Important caveats
Reasons your observed band may differ from the expected size.
ⓘCalculated mass138.1 kDa
ⓘLocalizationNucleus / Nucleus speckle
ⓘProcessing / PTMRecord-dependent
ⓘReactivityHuman

Sample controls for MECOM Western blot

🧪Use Adipose tissue as the first positive-control candidate; no defensible HPA Not detected tissue was available, so use a target knockdown/knockout negative control.
Positive control: Adipose tissue (High)
Negative control: Target knockdown/knockout
HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for MECOM

Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart on each tissue page — click any row's HPA link to view the source.

Positive expression · recommended positive controls

Tissue Cell type Level Evidence Source
Adipose tissue Reported tissue cells High Protein (HPA) HPA →
Breast Reported tissue cells High Protein (HPA) HPA →
Adrenal gland Reported tissue cells High Protein (HPA) HPA →
Appendix Reported tissue cells High Protein (HPA) HPA →

Undetected expression · recommended negative controls

Tissue Cell type Level Evidence Source

No HPA Not detected tissue is available; use a target knockdown/knockout negative control.

Section 1

What Is the Expected MECOM Western Blot Band Size?

Use the product-observed 138.1 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
138.1 kDaMatches the authoritative product WB observation.
138.1 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected MECOM appearancePlan around 138.1 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band138.1 kDa; use this as the primary experimental expectation.
Calculated mass138.1 kDa from UniProt Q03112; retain as context.
Gel selection8-10%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A04590-1.
Why is my band missing or off?
SituationLikely causeNext action
138.1 kDaMatches the authoritative product WB observation.Confirm with orthogonal controls and the linked product record.
Additional bandMay reflect processing, modification, or non-specific signal.Run a dilution series and compare positive/negative controls.
Weak signalTarget abundance or transfer may be limiting.Verify transfer, increase positive-control abundance, and bracket exposure.
Section 2

Real Curated MECOM Western Blot Protocols

Start with the molecular-weight rule, then compare verified publication-derived conditions.

Recommended Western blot protocol parameters
Sample / lysateAdipose tissue
Gel %8-10%
Load20-30 µg total protein per lane
TransferWet/tank, extended transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryA04590-1 at datasheet starting dilution
Primary incubationOvernight at 4 °C with gentle agitation
SecondarySpecies-matched HRP conjugate at validated dilution
Wash3 × 5 min in TBST
DetectionChemiluminescent substrate
ExposureBracket exposures to avoid saturation
Section 4

Advanced MECOM Western Blot Tips

Deeper troubleshooting and optimisation questions for MECOM, answered from its protein features.

Which band should guide the blot?
Use 138.1 kDa, the observation attached to the authoritative A04590-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 138.1 kDa expectation.
Which positive control should I start with?
Start with Adipose tissue, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use a target knockdown/knockout control when no HPA Not detected tissue is available.
Which gel should I use?
Use 8-10% consistently across the quick facts, protocol table, and poster.
What transfer method to use for MECOM Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A04590-1 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
Compare only the supplied target-verified records: PMC10130150, PMC4169276, PMC4716764.
Boster reagents

MECOM Western Blot Reagents

Human-reactive MECOM Western blot reagents with authoritative product imagery.

Real WB dataWestern blot validation image for MECOM using A04590-1; observed band 138.1 kDa
Anti-EVI1/MECOM Antibody Picoband®
Cat # A04590-1
Reactivity: Human

Only image-backed, WB-validated Human/Mouse/Rat recommendations from the prepared catalog evidence are shown.

Source: prepared picoband-wb product evidence; each card retains its own SKU, URL, observed band, and authoritative WB image.