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- Table of Contents
EEF2K · Western blot design guide
EEF2K Western Blot Planning Guide
Plan an EEF2K Western blot around the catalog-observed 82.1 kDa band, image-backed A02277-3 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
EEF2K 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 | 82.1 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 8-10% | |
| Positive control | Skin | |
| Negative control | Cerebral cortex |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 82.1 kDa |
| ⓘ | Localization | Not reported |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Rat |
Section 1
Real Curated EEF2K Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Skin |
| Gel % | 8-10% |
| 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 | A02277-3 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 EEF2K Western Blot Band Size?
Use the product-observed 82.1 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 82.1 kDa | Matches the authoritative product WB observation. |
| 82.1 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 EEF2K appearancePlan around 82.1 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 82.1 kDa; use this as the primary experimental expectation. |
| Calculated mass | 82.1 kDa from UniProt O00418; 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 A02277-3. |
| Situation | Likely cause | Next action |
|---|---|---|
| 82.1 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 EEF2K band
A02277-3 · EEF2K
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for EEF2K Western blot
🧪Use Skin as the first positive-control candidate and Cerebral cortex as the HPA Not detected negative candidate.
Positive control: Skin (Medium)
Negative control: Cerebral cortex (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for EEF2K
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 |
|---|---|---|---|---|
| Skin | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Breast | Reported tissue cells | Low | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | Low | Protein (HPA) | HPA → |
| Bone marrow | Reported tissue cells | Low | Protein (HPA) | HPA → |
Section 3
Advanced EEF2K Western Blot Tips
Deeper troubleshooting and optimisation questions for EEF2K, answered from its protein features.
Which band should guide the blot?
Use 82.1 kDa, the observation attached to the authoritative A02277-3 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 82.1 kDa expectation.
Which positive control should I start with?
Start with Skin, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Cerebral cortex as an orthogonal HPA Not detected candidate.
Which gel should I use?
Use 8-10% consistently across the quick facts, protocol table, and poster.
What transfer method to use for EEF2K Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A02277-3 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
EEF2K Western Blot Reagents
Human/Rat-reactive EEF2K Western blot reagents with authoritative product imagery.
Real WB data
Anti-EEF2K Antibody Picoband®
Cat # A02277-3
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 O00418
- Human Protein Atlas — EEF2K
- A02277-3 product record
- PMC11663172 — eEF2K alleviates doxorubicin-induced cardiotoxicity by inhibiting GSK3β and improving autophagy dysfunction (Cell biology and toxicology, 2024)
- PMC13042790 — Targeted Degradation of eEF2K by a Structure-Guided PROTAC Strategy for the Treatment of Triple-Negative Breast Cancer (Advanced science (Weinheim, Baden-Wurttemberg, Germany), 2026)
- PMC3580832 — Thermal injury activates the eEF2K-dependent eEF2 pathway in pediatric patients (JPEN. Journal of parenteral and enteral nutrition, 2012)