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- Table of Contents
MC4R · Western blot design guide
MC4R Western Blot Planning Guide
Plan a MC4R Western blot around the catalog-observed 36.9 kDa band, image-backed A00619-2 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
MC4R 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 | 36.9 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Bone marrow | |
| Negative control | Fallopian tube |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 36.9 kDa |
| ⓘ | Localization | Cell membrane / Cell projection, cilium membrane |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated MC4R Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Bone marrow |
| Gel % | 12-15% |
| Load | 20-30 µg total protein per lane |
| Transfer | Semi-dry, standard transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A00619-2 at datasheet starting dilution |
| Primary incubation | Overnight at 4 °C with gentle agitation |
| Secondary | Species-matched HRP conjugate at validated dilution |
| Wash | 3 × 5 min in TBST |
| Detection | Chemiluminescent substrate |
| Exposure | Bracket exposures to avoid saturation |
Section 2
What Is the Expected MC4R Western Blot Band Size?
Use the product-observed 36.9 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 36.9 kDa | Matches the authoritative product WB observation. |
| 36.9 kDa calculated | Use as UniProt context, not as a replacement observed band. |
| Unexpected additional signal | Do not assign identity without orthogonal positive/negative controls. |
💡Expected MC4R appearancePlan around 36.9 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 36.9 kDa; use this as the primary experimental expectation. |
| Calculated mass | 36.9 kDa from UniProt P32245; retain as context. |
| Gel selection | 12-15%; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with A00619-2. |
| Situation | Likely cause | Next action |
|---|---|---|
| 36.9 kDa | Matches the authoritative product WB observation. | Confirm with orthogonal controls and the linked product record. |
| Additional band | May reflect processing, modification, or non-specific signal. | Run a dilution series and compare positive/negative controls. |
| Weak signal | Target abundance or transfer may be limiting. | Verify transfer, increase positive-control abundance, and bracket exposure. |
Experiment evidence
Western blot images showing MC4R band
A00619-2 · MC4R
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MC4R Western blot
🧪Use Bone marrow as the first positive-control candidate and Fallopian tube as the HPA Not detected negative candidate.
Positive control: Bone marrow (High)
Negative control: Fallopian tube (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for MC4R
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
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Reported tissue cells | High | Protein (HPA) | HPA → |
| Kidney | Reported tissue cells | High | Protein (HPA) | HPA → |
| Nasopharynx | Reported tissue cells | High | Protein (HPA) | HPA → |
| Pancreas | Reported tissue cells | High | Protein (HPA) | HPA → |
Section 3
Advanced MC4R Western Blot Tips
Deeper troubleshooting and optimisation questions for MC4R, answered from its protein features.
Which band should guide the blot?
Use 36.9 kDa, the observation attached to the authoritative A00619-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 36.9 kDa expectation.
Which positive control should I start with?
Start with Bone marrow, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Fallopian tube 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 MC4R Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00619-2 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
MC4R Western Blot Reagents
Human/Mouse/Rat-reactive MC4R Western blot reagents with authoritative product imagery.
Real WB data
Anti-MC4-R/MC4R Antibody Picoband®
Cat # A00619-2
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
- UniProt P32245
- Human Protein Atlas — MC4R
- A00619-2 product record
- PMC3312751 — Melanocortin receptors in rat liver cells: change of gene expression and intracellular localization during acute-phase response (Histochemistry and cell biology, 2012)
- PMC11856147 — Melanocortin-4 Receptor Antagonism Inhibits Colorectal and Anaplastic Thyroid Cancer In Vitro and In Vivo (Journal of clinical medicine, 2025)
- PMC11825094 — Tonic ubiquitination of the central body weight regulator melanocortin receptor 4 (MC4R) promotes its constitutive exit from cilia (PLoS biology, 2025)
