This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
EIF3B / Eukaryotic translation initiation factor 3 subunit B · IHC design guide
Design Immunohistochemistry for EIF3B
Plan EIF3B chromogenic IHC on paraffin sections using the general cytoplasmic pattern and highly stained colon glandular cells as a reference (HPA tissue IHC). This guide covers fixation consistency, controls, antibody titration and cytoplasmic scoring.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
EIF3B Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | General cytoplasmic staining in glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04318-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA have medium consistency (HPA tissue IHC) | |
| Regulation | Stress recruits EIF3B to granules (UniProt) | |
| Isoform / epitope | 2 isoforms; verify epitope coverage (UniProt) |
Section 1
Recommended EIF3B IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A04318-1) is paired with three published EIF3B IHC tissue protocols (PMC11087095; PMC7439432; PMC6698483).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A04318-1) |
| Fixation | Image fixative and duration unreported (datasheet A04318-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A04318-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04318-1) |
| Primary antibody | Rabbit anti-EIF3B, 2-5 μg/ml (datasheet A04318-1) |
| Primary incubation | Overnight at 4 °C (datasheet A04318-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04318-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | EIF3B-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A04318-1); the published protocols use their own reported retrieval conditions.
Section 2
What Is the Expected EIF3B Staining Pattern?
EIF3B should show predominantly cytoplasmic staining in paraffin tissue sections (UniProt P55884: cytoplasm; HPA tissue IHC: general cytoplasmic expression). High staining is reported in glandular cells of the colon, duodenum, appendix, adrenal gland, and endometrium, among other listed cell types (HPA tissue IHC). HPA rates the tissue staining Approved, with medium consistency against RNA expression (HPA tissue IHC). EIF3B has no transmembrane segment (UniProt P55884 topology).
| Clear cytoplasmic stain in colonic or duodenal glandular cells. | This fits the reported High staining in those cells (HPA tissue IHC) and EIF3B's cytoplasmic location (UniProt P55884). Assess the signal against cell morphology and the controls on the same run (standard IHC practice). |
| Predominantly nuclear stain, with little cytoplasmic signal. | This departs from the general tissue IHC pattern (HPA tissue IHC: cytoplasmic). Investigate staining specificity and detection conditions (standard IHC practice). Limited nucleoplasmic localisation in ICC-IF is labelled uncertain, so nuclear signal alone is not established as a tissue IHC pattern (HPA subcellular). |
| Strong stain in cells outside the expected compartment or cell population. | Compare the stained cells with tissue morphology and HPA's cell-level observations (HPA tissue IHC). Cross-reactivity or endogenous detection activity are possible explanations, not diagnoses from appearance alone (standard IHC practice). HPA lists no negative tissue here, so an unlisted cell type is not automatically negative (HPA tissue IHC). |
| Broad haze obscures cell borders and cytoplasm. | Diffuse background prevents a reliable compartment call (standard IHC practice). Check the negative detection control, blocking, washes, and detection conditions before scoring (standard IHC practice). The expected pattern is cellular and generally cytoplasmic (HPA tissue IHC). |
| No signal in a previously positive, listed tissue. | A blank run conflicts with the reported High cell-level staining for that tissue (HPA tissue IHC). First verify tissue identity, run controls, retrieval, antibody incubation, and detection (standard IHC practice). HPA's Approved rating has medium RNA–staining consistency, so one slide should not be treated as a universal sensitivity test (HPA tissue IHC). |
💡Expected EIF3B appearanceCall a positive result when the relevant cells show discernible cytoplasmic staining, especially High staining in HPA-listed cell populations (HPA tissue IHC); dominant nuclear-only stain or diffuse cell-free haze warrants review (HPA tissue IHC; standard IHC practice).
| Tissue and cell selection | HPA reports low tissue specificity, with High staining in several epithelial or glandular populations and Purkinje cells; glial cells in hippocampus and smooth muscle cells are listed as Low (HPA tissue IHC). Use the named cell population when judging a control. |
| Cellular location and stress | EIF3B is cytoplasmic and can localise to stress granules following cellular stress (UniProt P55884). That conditional annotation does not establish that puncta should appear in routine paraffin sections (UniProt P55884). |
| Isoforms and epitope interpretation | UniProt lists 2 isoforms and no signal peptide, propeptide, or transmembrane segment (UniProt P55884). Those annotations alone do not identify which isoform an antibody detects or predict its performance after retrieval. |
| IHC antibody evidence | HPA lists IHC as Approved for HPA048983 and CAB017562; its tissue profile has medium consistency with RNA data (HPA antibodies; HPA tissue IHC). Treat agreement with the reported compartment and cell type as supporting evidence, then assess controls (standard IHC practice). |
| IF/ICC Q: where should EIF3B appear? | A: Mainly in the cytosol (HPA subcellular: supported); additional nucleoplasm is uncertain (HPA subcellular). UniProt reports stress-granule localisation following stress (UniProt P55884). These IF observations do not change the expected cytoplasmic tissue IHC pattern (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Listed positive cells are blank. | A failed run step or insufficient detectable signal is possible (standard IHC practice); HPA reports High staining in the named cells (HPA tissue IHC). | Confirm tissue and cell identity, then check retrieval, antibody incubation, and detection with run controls (standard IHC practice). Do not infer EIF3B-specific fixation sensitivity from these records. |
| Signal is nuclear-dominant. | The result differs from general cytoplasmic tissue staining (HPA tissue IHC); HPA calls the additional nucleoplasmic ICC-IF location uncertain (HPA subcellular). | Review cell morphology and controls; reassess staining and detection conditions before assigning localisation (standard IHC practice). |
| Unexpected cells stain strongly. | Cross-reactivity or endogenous detection activity is possible (standard IHC practice). HPA's tissue profile gives named cell-level observations, not a complete negative-cell list (HPA tissue IHC). | Check tissue anatomy and a negative detection control; investigate any signal that persists outside the expected cell pattern (standard IHC practice). |
| Background masks cytoplasmic detail. | Non-specific binding, inadequate washing, or detection background can obscure cellular staining (standard IHC practice). | Inspect the negative control and review blocking, washes, and detection conditions; score only where cytoplasm can be distinguished (standard IHC practice). |
| Hippocampal glia or smooth muscle look weak. | Low staining is reported for these cell populations (HPA tissue IHC). Weak signal there does not by itself indicate a failed run. | Compare with a listed High-staining cell population and the run controls before changing conditions (HPA tissue IHC; standard IHC practice). |
| Punctate cytoplasmic signal appears. | Stress-granule localisation is possible after cellular stress (UniProt P55884), but these records do not establish puncta as the routine paraffin IHC pattern. | Check whether general cytoplasmic staining and control behaviour remain credible; avoid calling puncta stress granules from IHC appearance alone (HPA tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing EIF3B staining
A04318-1 · IHCIHC analysis of EIF3B using anti-EIF3B antibody (A04318-1). EIF3B was detected in a paraffin-embedded section of human breast cancer tissue.
A04318-1 · IHCIHC analysis of EIF3B using anti-EIF3B antibody (A04318-1). EIF3B was detected in a paraffin-embedded section of human liver cancer tissue.
A04318-1 · IFIF analysis of EIF3B using anti-EIF3B antibody (A04318-1). EIF3B was detected in an immunocytochemical section of SiHa cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000106263-EIF3B/tissue/cerebral+cortex#imid_15282706
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000106263-EIF3B/tissue/colon#imid_15282624
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000106263-EIF3B/tissue/liver#imid_15282625
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000106263-EIF3B/tissue/pancreas#imid_15282584
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody CAB017562; green = EIF3B, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody CAB017562; green = EIF3B, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for EIF3B IHC & IF
🧪Run bronchus first; respiratory epithelial cells should stain strongly (HPA: High in respiratory epithelial cells). HPA detects EIF3B in all 45 scored tissues, so there is no validated negative tissue; use no-primary and isotype controls, and treat unstained acellular background as the within-slide negative reference rather than assuming any neighboring cell type is EIF3B-negative (HPA: no negative tissue rows).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: EIF3B is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show EIF3B in HEK293, MCF-7, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Run a no-primary, secondary-only section and a concentration-matched nonimmune rabbit IgG control alongside the test section (caption: rabbit primary antibody; standard IHC practice). Use an EIF3B knockout specimen or a validated immunizing-peptide block as a biological specificity control, and block endogenous peroxidase before HRP/DAB detection (standard IHC practice; caption: HRP/DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A04318-1 paraffin-section caption does not state the fixative (caption: fixative unreported). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required; the supplied evidence does not establish whether frozen sections or IF are easier (caption: heat-mediated EDTA retrieval; HPA: ICC-IF images in HEK293, MCF-7 and U2OS). In bronchus, endogenous peroxidase can produce misleading DAB signal if left unblocked (standard IHC practice; caption: HRP/DAB detection).
HPA tissue IHC evidence for EIF3B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: EIF3B is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced EIF3B IHC Tips
Troubleshoot EIF3B staining in paraffin sections by checking retrieval, cytoplasmic localisation, controls and scoring before interpreting changes in signal.
Which retrieval conditions should I start with for EIF3B paraffin-section IHC?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A04318-1). The selected paraffin-section example used that retrieval before incubation with 2 μg/ml primary antibody overnight at 4°C (datasheet A04318-1). If staining is weak, compare a slightly longer heating interval on matched sections while keeping cooling, antibody concentration and detection consistent (standard IHC practice). Examine tissue integrity alongside signal, because excessive heating can damage morphology and make cytoplasmic staining difficult to judge (standard IHC practice). Use a positive tissue control on each run; high staining in colon glandular cells provides one documented reference pattern (HPA: High in colon glandular cells).
How should I troubleshoot variable EIF3B staining across differently fixed sections?
The selected paraffin-section caption does not report a fixative, so target-specific fixation sensitivity is unknown (datasheet A04318-1). Record fixative, time to fixation, fixation duration and processing history for each specimen, then compare sections processed together (standard IHC practice). Keep retrieval at EDTA pH 8.0 and the primary antibody at 2 μg/ml during the initial comparison (datasheet A04318-1). Assess morphology and a positive control before attributing a weak result to fixation, since poor preservation or inconsistent processing can change apparent staining (standard IHC practice). Do not derive an EIF3B fixation requirement from its cytoplasmic location or phosphorylation sites (UniProt P55884: localisation and modified residues).
What EIF3B staining pattern should I expect in a paraffin section?
Look primarily for cytoplasmic staining, consistent with the general tissue-IHC pattern and EIF3B's annotated cytoplasmic location (HPA: general cytoplasmic expression; UniProt P55884: cytoplasm). Compare signal within the same cell type against a matched control section, rather than treating every stained structure as equivalent (standard IHC practice). Supported cytosolic localisation in cell imaging strengthens the compartment expectation, while nucleoplasmic localisation is marked uncertain (HPA: subcellular localisation). EIF3B can localise to stress granules following cellular stress, but punctate chromogenic staining alone cannot establish stress-granule identity (UniProt P55884: stress granules; standard IHC practice). Check glandular-cell staining in colon as a documented positive reference if that tissue is available (HPA: High in colon glandular cells).
Can this IHC stain distinguish EIF3B isoforms or modified epitopes?
EIF3B has 2 annotated isoforms, and the supplied antibody caption does not identify an epitope or establish isoform specificity (UniProt P55884: isoforms 1 and 2; datasheet A04318-1). Interpret a positive section as antibody-detected EIF3B signal rather than assigning it to a particular isoform (standard IHC interpretation). The protein contains an RNA recognition motif at residues 185–268 and annotated phosphorylation sites, including serines 78, 81, 83 and 85 (UniProt P55884: domains and modified residues). Those annotations do not show whether this antibody recognises a modified epitope (UniProt P55884: modified residues; datasheet A04318-1: epitope unspecified). Establish isoform or modification specificity with epitope documentation and orthogonal controls before making that claim (standard IHC practice).
How should I adapt the EIF3B localisation question to multiplex IF?
Treat IF/ICC as a separate assay: the selected antibody example documents paraffin-section chromogenic IHC, not an IF dilution or fixation condition (datasheet A04318-1). Multiplex EIF3B with a validated marker for the cell type being evaluated, and confirm that the two primary antibodies and detection channels can be distinguished (standard IF practice). Select fluorophores after checking tissue autofluorescence and single-channel controls, especially when evaluating faint cytosolic signal (standard IF practice; HPA: supported cytosolic localisation). EIF3B has no transmembrane segment and is annotated in the cytoplasm, so use controlled permeabilisation to access its intracellular epitope (UniProt P55884: topology and localisation; standard IF practice). Validate fixation and permeabilisation empirically with positive and negative controls; no target-specific IF conditions are supplied here (standard IF practice).
How can I separate EIF3B staining from diffuse chromogenic background?
Start with the documented 2 μg/ml primary antibody and 10% goat-serum block, then compare matched sections with and without primary antibody (datasheet A04318-1; standard IHC practice). The selected example uses peroxidase-linked secondary detection and DAB; quenching endogenous peroxidase is a general chromogenic IHC control, not evidence of an EIF3B-specific requirement (datasheet A04318-1; standard IHC practice). If no-primary sections stain, inspect blocking, washes, secondary-antibody binding and endogenous pigment before changing retrieval (standard IHC practice). Reduce overly broad signal by titrating primary antibody and detection conditions while retaining a positive control (standard IHC practice). Judge improvement by preserved cytoplasmic contrast and morphology, not by a darker DAB reaction alone (HPA: general cytoplasmic expression; standard IHC practice).
How should I score EIF3B IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic scoring region before comparing sections, since EIF3B shows general cytoplasmic tissue expression (HPA: tissue-IHC profile; standard IHC practice). Record the percentage of positive cells and staining intensity, or calculate an H-score as the sum of percentage at each intensity multiplied by its intensity grade (standard IHC practice). For spatial analyses, report positive-cell density per mm² of viable tissue and state how excluded regions were selected (standard IHC practice). Normalise comparisons to the same cell type, viable area, section handling and imaging or scoring settings (standard IHC practice). Include a shared positive control because the tissue-IHC profile has Approved reliability with medium consistency against RNA expression (HPA: reliability).
When is an unexpected EIF3B-positive area likely to be artefactual?
A convincing result should show reproducible staining in the expected cytoplasmic compartment and in identifiable viable cells (HPA: general cytoplasmic expression; standard IHC practice). Compare the questioned area with a no-primary control and a known positive section; high staining in colon glandular cells is documented (standard IHC practice; HPA: High in colon glandular cells). Treat isolated nuclear-only signal cautiously because cytosol is supported in cell imaging whereas nucleoplasmic localisation is uncertain (HPA: subcellular localisation). Inspect section edges and necrotic regions for uneven or nonspecific DAB deposition, and check endogenous peroxidase with an appropriate control (standard IHC practice). A change in EIF3B IHC intensity alone does not establish altered translation-initiation activity (UniProt P55884: eIF-3 function; standard IHC interpretation).
Boster reagents
Best EIF3B / Eukaryotic translation initiation factor 3 subunit B IHC Antibodies
A04318-1 has real IHC data from human paraffin sections and IF data from SiHa cells (catalog image captions); its listed reactivity covers human, mouse and rat (catalog).
Real IHC data
Anti-EIF3B Antibody ®
Cat # A04318-1
A04318-1 will render with an IHC figure from a human breast cancer paraffin section; its separate IF caption documents SiHa cells (catalog image captions). M04318 will render without an IHC or IF figure; ICC/IF is listed among its applications, with human, mouse and rat reactivity (catalog).
Which to pick: Choose A04318-1 for tissue IHC: its own caption documents staining in a human breast cancer paraffin section after EDTA pH 8.0 retrieval at 2 μg/ml; the fixative is unreported (A04318-1 IHC caption). For IF/ICC, A04318-1 has a SiHa cell IF image at 5 μg/ml, while M04318 is a rabbit monoclonal with ICC/IF listed but no supplied IF image (catalog). Both list human, mouse and rat reactivity, but the supplied tissue IHC images document human samples only (catalog; A04318-1 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P55884 (EIF3B_HUMAN, Eukaryotic translation initiation factor 3 subunit B).
- Human Protein Atlas. EIF3B tissue IHC expression (reliability: Approved).
- Human Protein Atlas. EIF3B subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the nucleoplasm..
- Human Protein Atlas. EIF3B antibody validation summary (2 antibodies).
- Eukaryotic Translation Initiation Factor 3b is both a Promising Prognostic Biomarker and a Potential Therapeutic Target for Patients with Clear Cell Renal Cell Carcinoma. Journal of Cancer 2017 — PMC5604456.
- EIF3B stabilizes PCNA by counteracting SYVN1-mediated ubiquitination to serve as a promotor in cholangiocarcinoma. Aging 2024 — PMC11087095.
- Eukaryotic initiation factor 3B is overexpressed and correlates with larger tumor size, advanced FIGO stage, and shorter overall survival in epithelial ovarian cancer patients. Journal of clinical laboratory analysis 2020 — PMC7439432.
- Downregulation of eukaryotic translation initiation factor 3b inhibited proliferation and metastasis of gastric cancer. Cell death & disease 2019 — PMC6698483.
- PubMed PMID:9388245 — UniProt-cited evidence.
- PubMed PMID:8995410 — UniProt-cited evidence.
- PubMed PMID:12853948 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for EIF3B (Eukaryotic translation initiation factor 3 subunit B). Expected localisation Cytoplasmic staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04318-1). Validated IHC antibody: A04318-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A04318-1); 2. Fixation: Image fixative and duration unreported (datasheet A04318-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A04318-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04318-1); 8. Primary antibody: Rabbit anti-EIF3B, 2-5 μg/ml (datasheet A04318-1); 9. Primary incubation: Overnight at 4 °C (datasheet A04318-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA have medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; verify epitope coverage (UniProt); Regulation: Stress recruits EIF3B to granules (UniProt). Evidence: UniProt P55884, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A04318-1, PMC11087095, PMC7439432. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/EIF3B/immunohistochemistry-ihc-guide.