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- Table of Contents
HINT1 · Western blot design guide
HINT1 Western Blot Planning Guide
Plan a HINT1 Western blot around the catalog-observed 13.8 kDa band, image-backed PB9632 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
HINT1 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 | 13.8 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 15% | |
| Positive control | Kidney | |
| Negative control | Heart muscle |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 13.8 kDa |
| ⓘ | Localization | Cytoplasm / Nucleus |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated HINT1 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Epididymis |
| Gel % | 15% |
| Load | 20-30 µg total protein per lane |
| Transfer | Semi-dry, short transfer |
| Membrane | 0.2 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | PB9632 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 HINT1 Western Blot Band Size?
Use the product-observed 13.8 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 13.8 kDa | Matches the authoritative product WB observation. |
| 13.8 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 HINT1 appearancePlan around 13.8 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 13.8 kDa; use this as the primary experimental expectation. |
| Calculated mass | 13.8 kDa from UniProt P49773; retain as context. |
| Gel selection | 15%; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with PB9632. |
| Situation | Likely cause | Next action |
|---|---|---|
| 13.8 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 HINT1 band
PB9632 · HINT1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HINT1 Western blot
🧪Use Kidney as the first positive-control candidate and Heart muscle as the HPA Not detected negative candidate.
Positive control: Kidney (High)
Negative control: Heart muscle (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for HINT1
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 |
|---|---|---|---|---|
| Kidney | Reported tissue cells | High | Protein (HPA) | HPA → |
| Fallopian tube | Reported tissue cells | High | Protein (HPA) | HPA → |
| Ovary | Reported tissue cells | High | Protein (HPA) | HPA → |
| Gallbladder | Reported tissue cells | High | Protein (HPA) | HPA → |
Section 3
Advanced HINT1 Western Blot Tips
Deeper troubleshooting and optimisation questions for HINT1, answered from its protein features.
Which band should guide the blot?
Use 13.8 kDa, the observation attached to the authoritative PB9632 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 13.8 kDa expectation.
Which positive control should I start with?
Start with Kidney, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Heart muscle as an orthogonal HPA Not detected candidate.
Which gel should I use?
Use 15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for HINT1 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should PB9632 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
HINT1 Western Blot Reagents
Human/Mouse/Rat-reactive HINT1 Western blot reagents with authoritative product imagery.
Real WB data
Anti-HINT1 Antibody Picoband®
Cat # PB9632
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 P49773
- Human Protein Atlas — HINT1
- PB9632 product record
- PMC12886079 — S-nitrosylation of HINT1 in macrophages aggravates foam cell formation and atherosclerosis (Redox biology, 2026)
- PMC8080686 — Deacetylation by SIRT1 promotes the tumor-suppressive activity of HINT1 by enhancing its binding capacity for β-catenin or MITF in colon cancer and melanoma cells (Experimental & molecular medicine, 2020)
- PMC10940622 — Histidine re-sensitizes pediatric acute lymphoblastic leukemia to 6-mercaptopurine through tetrahydrofolate consumption and SIRT5-mediated desuccinylation (Cell death & disease, 2024)