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- Table of Contents
HCCS · Western blot design guide
HCCS Western Blot Planning Guide
Plan a HCCS Western blot around the catalog-observed 30.6 kDa band, image-backed A00924-2 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
HCCS 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 | 30.6 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Parathyroid gland | |
| Negative control | Soft tissue |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 30.6 kDa |
| ⓘ | Localization | Mitochondrion inner membrane / Membrane |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated HCCS Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Oral mucosa |
| 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 | A00924-2 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 HCCS Western Blot Band Size?
Use the product-observed 30.6 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 30.6 kDa | Matches the authoritative product WB observation. |
| 30.6 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 HCCS appearancePlan around 30.6 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 30.6 kDa; use this as the primary experimental expectation. |
| Calculated mass | 30.6 kDa from UniProt P53701; 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 A00924-2. |
| Situation | Likely cause | Next action |
|---|---|---|
| 30.6 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 HCCS band
A00924-2 · HCCS
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HCCS Western blot
🧪Use Parathyroid gland as the first positive-control candidate and Soft tissue as the HPA Not detected negative candidate.
Positive control: Parathyroid gland (High)
Negative control: Soft tissue (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for HCCS
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 |
|---|---|---|---|---|
| Parathyroid gland | Reported tissue cells | High | Protein (HPA) | HPA → |
| Oral mucosa | Reported tissue cells | High | Protein (HPA) | HPA → |
| Colon | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Adrenal gland | Reported tissue cells | Medium | Protein (HPA) | HPA → |
Section 3
Advanced HCCS Western Blot Tips
Deeper troubleshooting and optimisation questions for HCCS, answered from its protein features.
Which band should guide the blot?
Use 30.6 kDa, the observation attached to the authoritative A00924-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 30.6 kDa expectation.
Which positive control should I start with?
Start with Parathyroid gland, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Soft tissue 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 HCCS Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00924-2 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
HCCS Western Blot Reagents
Human/Mouse/Rat-reactive HCCS Western blot reagents with authoritative product imagery.
Real WB data
Anti-HCCS Antibody Picoband®
Cat # A00924-2
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 P53701
- Human Protein Atlas — HCCS
- A00924-2 product record
- PMC4139152 — Conserved residues of the human mitochondrial holocytochrome c synthase mediate interactions with heme (Biochemistry, 2014)
- PMC13225780 — Multi-omics landscape and functional validation of HCCS in breast cancer: from pan-cancer immunometabolic characterization to regulating tumor proliferation (Frontiers in genetics, 2026)
