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- Table of Contents
HSPA9 · Western blot design guide
HSPA9 Western Blot Planning Guide
Plan a HSPA9 Western blot around the catalog-observed 73.7 kDa band, image-backed PA1789 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
HSPA9 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 | 73.7 kDa observed | |
| Observed band | 73.7 kDa observed | |
| Gel | 12-15% | |
| Positive control | Adrenal gland | |
| Negative control | Skeletal muscle |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 73.7 kDa |
| ⓘ | Localization | Mitochondrion / Nucleus, nucleolus |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated HSPA9 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Adrenal gland |
| 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 | PA1789 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 HSPA9 Western Blot Band Size?
Use the product-observed 73.7 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 73.7 kDa | Matches the authoritative product WB observation. |
| 73.7 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 HSPA9 appearancePlan around 73.7 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 73.7 kDa; use this as the primary experimental expectation. |
| Calculated mass | 73.7 kDa from UniProt P38646; 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 PA1789. |
| Situation | Likely cause | Next action |
|---|---|---|
| 73.7 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 HSPA9 band
PA1789 · HSPA9Observed 73.7 kDa · product-specific WB image
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HSPA9 Western blot
🧪Use Adrenal gland as the first positive-control candidate and retain an orthogonal negative control.
Positive control: Adrenal gland (High)
Negative control: Skeletal muscle (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for HSPA9
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 |
|---|---|---|---|---|
| Adrenal gland | Reported tissue cells | High | Protein (HPA) | HPA → |
| Breast | Reported tissue cells | High | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | High | Protein (HPA) | HPA → |
| Bone marrow | Reported tissue cells | High | Protein (HPA) | HPA → |
| Adipose tissue | Reported tissue cells | Low | Protein (HPA) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Skeletal muscle | Reported tissue cells | Not detected | Protein (HPA) | HPA → |
Section 3
Advanced HSPA9 Western Blot Tips
Deeper troubleshooting and optimisation questions for HSPA9, answered from its protein features.
Which band should guide the blot?
Use 73.7 kDa, the observation attached to the authoritative PA1789 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 73.7 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 Skeletal muscle 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 HSPA9 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should PA1789 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
HSPA9 Western Blot Reagents
Human/Mouse/Rat-reactive HSPA9 Western blot reagents with authoritative product imagery.
Real WB data
Anti-Grp75/HSPA9 Antibody Picoband®
Cat # PA1789
Real WB data
Anti-Grp75 Antibody Picoband® (monoclonal, 4I9)
Cat # M02561-2
Real WB data
Anti-Grp75/HSPA9 Antibody Picoband®
Cat # PB9642
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.