ADAM12 · Western blot design guide

ADAM12 Western Blot Planning Guide

Plan an ADAM12 Western blot around the catalog-observed 99.5 kDa band, image-backed A02264-2 evidence, HPA controls, and verified protocol records.

Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
Western blot protocol sheet for ADAM12 (ADAM12): expected band 99.5 kDa, antibody A02264-2, and guide-derived SDS-PAGE protocol steps
ADAM12 Western blot protocol sheet — expected band 99.5 kDa, antibody A02264-2, controls and PMC citations. Open the full ADAM12 WB guide →

ADAM12 Western Blot Experimental Design Guide

Expected bands, documented protocol parameters, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before running
Expected band 99.5 kDa
Observed band Not reported — verify product WB image
Gel 8-10%
Positive control ⓘ Testis
Negative control ⓘ Adrenal gland
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 99.5 kDa
ⓘ Localization Cell membrane / Secreted
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human / Mouse / Rat
Section 1

Real Curated ADAM12 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysatePlacenta
Gel %8-10%
Load20-30 µg total protein per lane
TransferSemi-dry, standard transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryA02264-2 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 2

What Is the Expected ADAM12 Western Blot Band Size?

Use the product-observed 99.5 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
99.5 kDaMatches the authoritative product WB observation.
99.5 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected ADAM12 appearancePlan around 99.5 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band99.5 kDa; use this as the primary experimental expectation.
Calculated mass99.5 kDa from UniProt O43184; retain as context.
Gel selection8-10%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A02264-2.
Why is my band missing or off?
SituationLikely causeNext action
99.5 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.

Sample controls for ADAM12 Western blot

🧪Use Testis as the first positive-control candidate and Adrenal gland as the HPA Not detected negative candidate.
Positive control: Testis (High)
Negative control: Adrenal gland (Not detected)
HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for ADAM12

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

Positive expression · recommended positive controls

TissueCell typeLevelEvidenceSource
Testis Reported tissue cells High Protein (HPA) HPA →
Placenta Reported tissue cells High Protein (HPA) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Adrenal gland Reported tissue cells Not detected Protein (HPA) HPA →
Appendix Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced ADAM12 Western Blot Tips

Deeper troubleshooting and optimisation questions for ADAM12, answered from its protein features.

Which band should guide the blot?
Use 99.5 kDa, the observation attached to the authoritative A02264-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 99.5 kDa expectation.
Which positive control should I start with?
Start with Testis, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Adrenal gland as an orthogonal HPA Not detected candidate.
Which gel should I use?
Use 8-10% consistently across the quick facts, protocol table, and poster.
What transfer method to use for ADAM12 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A02264-2 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
No verified publication protocol was supplied; use only the deterministic recommended protocol.
Boster reagents

ADAM12 Western Blot Reagents

Human/Mouse/Rat-reactive ADAM12 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for ADAM12 using A02264-2; observed band 99.5 kDa
Anti-ADAM12 Antibody Picoband®
Cat # A02264-2

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.

References

  1. UniProt O43184
  2. Human Protein Atlas — ADAM12
  3. A02264-2 product record
  4. PMC7115740 — ADAM12 silencing promotes cellular apoptosis by activating autophagy in choriocarcinoma cells (International journal of oncology, 2020)
  5. PMC5668165 — ADAM12 Is a Novel Regulator of Tumor Angiogenesis via STAT3 Signaling (Molecular cancer research : MCR, 2017)