MBD1 · Western blot design guide

MBD1 Western Blot Planning Guide

Plan a MBD1 Western blot around the catalog-observed 66.6 kDa band, image-backed A02336-1 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 MBD1 (MBD1): expected band 66.6 kDa, antibody A02336-1, and guide-derived SDS-PAGE protocol steps
MBD1 Western blot protocol sheet — expected band 66.6 kDa, antibody A02336-1, controls and PMC citations. Open the full MBD1 WB guide →

MBD1 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 66.6 kDa
Observed band Not reported — verify product WB image
Gel 12-15%
Positive control ⓘ Adrenal gland
Negative control ⓘ Seminal vesicle
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 66.6 kDa
ⓘ Localization Nucleus / Nucleus matrix
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human / Mouse / Rat
Section 1

Real Curated MBD1 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateAdrenal gland
Gel %12-15%
Load20-30 µg total protein per lane
TransferSemi-dry, standard transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryA02336-1 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 2

What Is the Expected MBD1 Western Blot Band Size?

Use the product-observed 66.6 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
66.6 kDaMatches the authoritative product WB observation.
66.6 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected MBD1 appearancePlan around 66.6 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band66.6 kDa; use this as the primary experimental expectation.
Calculated mass66.6 kDa from UniProt Q9UIS9; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A02336-1.
Why is my band missing or off?
SituationLikely causeNext action
66.6 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 MBD1 Western blot

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

HPA tissue expression evidence for MBD1

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
Adrenal gland Reported tissue cells Medium Protein (HPA) HPA →
Appendix Reported tissue cells Medium Protein (HPA) HPA →
Bone marrow Reported tissue cells Medium Protein (HPA) HPA →
Bronchus Reported tissue cells Medium Protein (HPA) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Seminal vesicle Reported tissue cells Not detected Protein (HPA) HPA →
Smooth muscle Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced MBD1 Western Blot Tips

Deeper troubleshooting and optimisation questions for MBD1, answered from its protein features.

Which band should guide the blot?
Use 66.6 kDa, the observation attached to the authoritative A02336-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 66.6 kDa expectation.
Which positive control should I start with?
Start with Adrenal gland, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Seminal vesicle as an orthogonal HPA Not detected candidate.
Which gel should I use?
Use 12-15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for MBD1 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A02336-1 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
No verified publication protocol was supplied; use only the deterministic recommended protocol.
Boster reagents

MBD1 Western Blot Reagents

Human/Mouse/Rat-reactive MBD1 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for MBD1 using A02336-1; observed band 66.6 kDa
Anti-MBD1 Antibody Picoband®
Cat # A02336-1

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 Q9UIS9
  2. Human Protein Atlas — MBD1
  3. A02336-1 product record
  4. PMC10531752 — Effect of Fetal Bovine Serum or Basic Fibroblast Growth Factor on Cell Survival and the Proliferation of Neural Stem Cells: The Influence of Homocysteine Treatment (International journal of molecular sciences, 2023)
  5. PMC13391528 — [MBD1 knockdown inhibits proliferation, migration and angiogenesis of human umbilical vein endothelial cells in vitro] (Nan fang yi ke da xue xue bao = Journal of Southern Medical University, 2026)