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- Table of Contents
HDAC5 · Western blot design guide
HDAC5 Western Blot Planning Guide
Plan a HDAC5 Western blot around the catalog-observed 122 kDa band, image-backed A01230-5 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
HDAC5 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 | 122 kDa observed | |
| Observed band | 122 kDa observed | |
| Gel | 8-10% | |
| Positive control | Colon | |
| Negative control | Use knockout/knockdown or orthogonal negative |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 122 kDa |
| ⓘ | Localization | Nucleus / Cytoplasm |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated HDAC5 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Appendix |
| Gel % | 8-10% |
| Load | 20-30 µg total protein per lane |
| Transfer | Wet/tank, extended transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A01230-5 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 HDAC5 Western Blot Band Size?
Use the product-observed 122 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 122 kDa | Matches the authoritative product WB observation. |
| 122 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 HDAC5 appearancePlan around 122 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 122 kDa; use this as the primary experimental expectation. |
| Calculated mass | 122 kDa from UniProt Q9UQL6; retain as context. |
| Gel selection | 8-10%; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with A01230-5. |
| Situation | Likely cause | Next action |
|---|---|---|
| 122 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 HDAC5 band
A01230-5 · HDAC5Observed 122 kDa · product-specific WB image
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HDAC5 Western blot
🧪Use Colon as the first positive-control candidate and retain an orthogonal negative control.
Positive control: Colon (High)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for HDAC5
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 |
|---|---|---|---|---|
| Colon | Reported tissue cells | High | Protein (HPA) | HPA → |
| Salivary gland | Reported tissue cells | High | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | High | Protein (HPA) | HPA → |
| Bone marrow | Reported tissue cells | High | Protein (HPA) | HPA → |
| Heart muscle | Reported tissue cells | Low | Protein (HPA) | HPA → |
| Skeletal muscle | Reported tissue cells | Low | Protein (HPA) | HPA → |
Undetected expression · recommended negative controls
The available HPA profile lists no reliably-negative tissue for HDAC5 (a broadly-expressed target). Use a HDAC5 knockdown (siRNA/shRNA) or knockout lysate as the negative control and confirm signal loss versus the parental lysate.
| Tissue | Cell type | Level | Evidence | Source |
|---|
Section 3
Advanced HDAC5 Western Blot Tips
Deeper troubleshooting and optimisation questions for HDAC5, answered from its protein features.
Which band should guide the blot?
Use 122 kDa, the observation attached to the authoritative A01230-5 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 122 kDa expectation.
Which positive control should I start with?
Start with Colon, 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 HDAC5 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A01230-5 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
HDAC5 Western Blot Reagents
Human/Mouse/Rat-reactive HDAC5 Western blot reagents with authoritative product imagery.
Real WB data
Anti-HDAC5 Antibody Picoband®
Cat # A01230-5
Real WB data
Anti-HDAC5 Antibody Picoband®
Cat # A01230-6
Real WB data
Anti-Histone deacetylase 5 HDAC5 Antibody Picoband®
Cat # PA2180
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.
