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- Table of Contents
EPO · Western blot design guide
EPO Western Blot Planning Guide
Plan a EPO Western blot around the catalog-observed 21.3 kDa band, image-backed RP1004 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
EPO 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 | 21.3 kDa observed | |
| Observed band | 21.3 kDa observed | |
| Gel | 12-15% | |
| Positive control | Breast | |
| Negative control | Breast |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 21.3 kDa |
| ⓘ | Localization | Secreted |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated EPO Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Adipose tissue |
| Gel % | 12-15% |
| Load | 20-30 µg total protein per lane |
| Transfer | Semi-dry, short transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | RP1004 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 EPO Western Blot Band Size?
Use the product-observed 21.3 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 21.3 kDa | Matches the authoritative product WB observation. |
| 21.3 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 EPO appearancePlan around 21.3 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 21.3 kDa; use this as the primary experimental expectation. |
| Calculated mass | 21.3 kDa from UniProt P01588; 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 RP1004. |
| Situation | Likely cause | Next action |
|---|---|---|
| 21.3 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 EPO band
RP1004 · EPOObserved 21.3 kDa · product-specific WB image
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for EPO Western blot
🧪Use Breast as the first positive-control candidate and retain an orthogonal negative control.
Positive control: Breast (Not detected)
Negative control: Breast (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for EPO
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 |
|---|
Undetected expression · recommended negative controls
The Human Protein Atlas exposes no structured IHC tissue profile for EPO, so tissue-level positive/negative controls cannot be listed here. Use a documented EPO-positive lysate (recombinant EPO or a known-expressing line) as the positive control and a EPO knockdown/knockout lysate as the negative control; confirm against the antibody datasheet.
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Breast | Reported tissue cells | Not detected | Protein (HPA) | HPA → |
| Adipose tissue | Reported tissue cells | Not detected | Protein (HPA) | HPA → |
| Adrenal gland | Reported tissue cells | Not detected | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | Not detected | Protein (HPA) | HPA → |
| Adrenal gland | Reported tissue cells | Not detected | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | Not detected | Protein (HPA) | HPA → |
Section 3
Advanced EPO Western Blot Tips
Deeper troubleshooting and optimisation questions for EPO, answered from its protein features.
Which band should guide the blot?
Use 21.3 kDa, the observation attached to the authoritative RP1004 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 21.3 kDa expectation.
Which positive control should I start with?
Start with Breast, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Breast 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 EPO Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should RP1004 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
EPO Western Blot Reagents
Human/Mouse/Rat-reactive EPO Western blot reagents with authoritative product imagery.
Real WB data
Anti-EPO/Erythropoietin Antibody Picoband®
Cat # RP1004
Real WB data
Anti-Erythropoietin EPO Antibody Picoband®
Cat # A00484-2
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.