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- Table of Contents
EIF3B · Western blot design guide
EIF3B Western Blot Planning Guide
Plan a EIF3B Western blot around the catalog-observed 92.5 kDa band, image-backed A04318-1 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
EIF3B 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 | 92.5 kDa observed | |
| Observed band | 92.5 kDa observed | |
| Gel | 8-10% | |
| Positive control | Cervix | |
| Negative control | Use knockout/knockdown or orthogonal negative |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 92.5 kDa |
| ⓘ | Localization | Cytoplasm / Cytoplasm, Stress granule |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated EIF3B Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Adrenal gland |
| 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 | A04318-1 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 EIF3B Western Blot Band Size?
Use the product-observed 92.5 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 92.5 kDa | Matches the authoritative product WB observation. |
| 92.5 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 EIF3B appearancePlan around 92.5 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 92.5 kDa; use this as the primary experimental expectation. |
| Calculated mass | 92.5 kDa from UniProt P55884; 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 A04318-1. |
| Situation | Likely cause | Next action |
|---|---|---|
| 92.5 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 EIF3B band
A04318-1 · EIF3B
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for EIF3B Western blot
🧪Use Cervix as the first positive-control candidate and retain an orthogonal negative control.
Positive control: Cervix (High)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for EIF3B
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 |
|---|---|---|---|---|
| Cervix | Reported tissue cells | High | Protein (HPA) | HPA → |
| Adrenal gland | Reported tissue cells | High | Protein (HPA) | HPA → |
| Cerebellum | Reported tissue cells | High | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | High | Protein (HPA) | HPA → |
| Smooth muscle | Reported tissue cells | Low | Protein (HPA) | HPA → |
| Hippocampus | Reported tissue cells | Low | Protein (HPA) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|
Section 3
Advanced EIF3B Western Blot Tips
Deeper troubleshooting and optimisation questions for EIF3B, answered from its protein features.
Which band should guide the blot?
Use 92.5 kDa, the observation attached to the authoritative A04318-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 92.5 kDa expectation.
Which positive control should I start with?
Start with Cervix, 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 EIF3B Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A04318-1 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
EIF3B Western Blot Reagents
Human/Mouse/Rat-reactive EIF3B Western blot reagents with authoritative product imagery.
Real WB data
Anti-EIF3B Antibody Picoband®
Cat # A04318-1
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 P55884
- Human Protein Atlas — EIF3B
- A04318-1 product record
- PMC11380452 — FOXA1-dependent PUS1 regulates EIF3b stability in a non-enzymatic pathway mediating prostate cancer bone metastasis (International journal of biological sciences, 2024)
- PMC2666973 — Phosphorylation of the eukaryotic initiation factor 3f by cyclin-dependent kinase 11 during apoptosis (FEBS letters, 2009)
- PMC12998141 — EIF3B regulates the cell cycle of lung adenocarcinoma cells by activating the GTSE1 mediated ERK/MAPK pathway (Respiratory research, 2026)