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EIF3F / Eukaryotic translation initiation factor 3 subunit F · Western blot design guide
Design a Western Blot for EIF3F
Source-linked EIF3F Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-EIF3F WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
EIF3F Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~37.6 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked EIF3F Western Blot Protocol Options
The A30682 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | various cells (catalog A30682) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A30682; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected EIF3F Western Blot Band Size?
EIF3F is predicted at 37.6 kDa; annotated modifications and complex membership could affect appearance, but distinct migration effects are not demonstrated.
| Band near 37.6 kDa | Consistent with full-length EIF3F; confirm band identity with antibody controls. |
| Band near 37.6 kDa in cytoplasmic lysate | Consistent with EIF3F's annotated cytoplasmic location. |
| Close doublet near 37.6 kDa | Could reflect annotated phosphorylation, but a visible shift is not established. |
| Higher band under conditions that preserve complexes | Could reflect EIF3F in the eIF3 complex; confirm its identity. |
💡Expected EIF3F appearanceEIF3F has a predicted mass of 37.6 kDa and no supplied empirical band size; look near 37.6 kDa in cytoplasmic lysate and confirm identity with antibody controls.
| Predicted EIF3F mass | Places the full-length protein near 37.6 kDa. |
| N-acetylalanine at residue 2 | No visible migration effect is established. |
| Phosphoserine at residue 46 by CDK11 in vitro | A migration shift is not established for cellular blots. |
| N6-acetyllysine at residue 238 | No visible migration effect is established. |
| Phosphoserine at residue 258 | A migration shift is possible but not demonstrated. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | EIF3F is cytoplasmic; absence may reflect poor cytoplasmic recovery or detection. | Check cytoplasmic extraction, loading, and antibody performance. |
| Band higher than expected | EIF3F participates in the eIF3 complex, which may persist if denaturation is incomplete. | Check denaturation and confirm band identity with antibody controls. |
| Band lower than expected | No signal peptide or propeptide cleavage is annotated; the smaller band's identity is uncertain. | Check sample integrity and confirm band identity. |
| Multiple bands | Only one isoform is annotated; additional bands have no established assignment. | Use peptide blocking or another antibody to assess specificity. |
| Weak or no signal | EIF3F may be insufficiently detected in the prepared lysate. | Check loading, transfer, and antibody performance. |
Experiment evidence
Western blot images showing EIF3F band
A30682 · EIF3F
A30682 · EIF3F
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for EIF3F Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for EIF3F in Western blot, you can use adrenal gland lysate, a high-expression HPA tissue.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Adipose tissue is reported as not detected, but confirm the negative result in your lysate.
HPA tissue expression evidence for EIF3F
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.
Higher expression tissues · candidate positive controls from IHC
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Endometrium | glandular cells | High | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Low | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Low | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced EIF3F Western Blot Tips
Deeper troubleshooting and optimisation questions for EIF3F, answered from its protein features.
How should EIF3F band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Are alternative EIF3F isoforms expected?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no basis for assigning multiple bands to EIF3F isoforms.
Which EIF3F phosphorylation sites are annotated?
PTM · UniProt lists phosphoserine at residue 46, attributed to CDK11 in vitro, and phosphoserine at residue 258. These are UniProt coordinates and may differ from paper or antibody numbering. Neither annotation establishes a detectable band shift.
UniProt lists N-acetylalanine at residue 2 and N6-acetyllysine at residue 238. Check coordinate conventions when comparing antibody or paper descriptions. These annotations do not establish distinct Western blot bands.
UniProt lists N-acetylalanine at residue 2 and N6-acetyllysine at residue 238. Check coordinate conventions when comparing antibody or paper descriptions. These annotations do not establish distinct Western blot bands.
Does this guide establish induction of EIF3F?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
How should transfer be checked for EIF3F?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A30682 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should EIF3F be quantified on a Western blot?
Quantitation · EIF3F is annotated in the cytoplasm and as a component of the eIF-3 complex. Compare the same candidate band across comparable samples, using a consistent loading reference and exposures within the measurement range. The record does not establish modification-specific bands for separate quantification.
How should an EIF3F band compare with predicted mass?
Interpretation · EIF3F has a predicted mass of 37.6 kDa, but no observed band position is supplied. Use 37.6 kDa as a reference, not an exact expected migration position. The annotated modifications alone do not establish a visible shift or explain any difference from predicted mass.
The record lists one isoform, no glycosylation sites, and no signal peptide or propeptide. Its four modified residues do not, by themselves, explain extra bands. Verify the identity of any unexpected band before assigning it to EIF3F or a particular modification.
The record lists one isoform, no glycosylation sites, and no signal peptide or propeptide. Its four modified residues do not, by themselves, explain extra bands. Verify the identity of any unexpected band before assigning it to EIF3F or a particular modification.
Boster reagents
EIF3F Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-EIF3F/Eif3 Epsilon Antibody
Cat # A30682
The catalog reports A30682, an anti-EIF3F antibody with reported human, mouse, and rat reactivity. A Western blot image is available, described as analysis of various cells; the supplied evidence does not identify the cell types or establish validation across all three species.
Which to pick: A30682 is the only listed EIF3F option and has a Western blot image. Check that its reported reactivity matches your sample; the image caption does not specify which cells or species were tested.
Source: BosterBio EIF3F gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O00303.
- Human Protein Atlas. EIF3F tissue expression.
- PMC4619320 — target-verified WB comparison
- PMC13459262 — target-verified WB comparison
- PMC7599166 — target-verified WB comparison