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EIF3C / Eukaryotic translation initiation factor 3 subunit C · IHC design guide
Design Immunohistochemistry for EIF3C
Plan EIF3C chromogenic IHC in paraffin sections with a catalog antibody range of 2–5 μg/ml (datasheet A06470-1). Assess cytoplasmic staining against the tissue pattern and consider HPA’s multi-gene targeting caution when scoring (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
EIF3C 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 | General cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | High cytoplasmic signal in glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06470-1) | |
| Positive control | Appendix+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 | Multi-gene protein targeting may confound scoring (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unreported (UniProt) |
Section 1
Recommended EIF3C IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A06470-1) is accompanied by four published EIF3C tissue IHC protocols (PMC6331022; PMC7144925; PMC6389005; PMC9492341).
| Sample | Paraffin-embedded human lymphoma tissue; fixative not specified (datasheet A06470-1) |
| Fixation | Image fixative and duration unreported (datasheet A06470-1); 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 A06470-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06470-1) |
| Primary antibody | Rabbit anti-EIF3C, 2-5 μg/ml (datasheet A06470-1) |
| Primary incubation | Overnight at 4 °C (datasheet A06470-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A06470-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | EIF3C-positive staining in glandular cells of appendix (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 (datasheet A06470-1). If needed, compare citrate pH 6 (PMC7144925) or pH 9 retrieval (PMC6331022).
Section 2
What Is the Expected EIF3C Staining Pattern?
EIF3C should show predominantly cytoplasmic staining in paraffin sections (UniProt Q99613: cytoplasm; HPA tissue IHC: general cytoplasmic expression). Expect strong signal in appendix, breast, colon, and duodenal glandular cells, among other HPA high-staining cells (HPA tissue IHC). HPA rates its tissue pattern Supported, with medium agreement between staining and RNA data, and cautions that the antibodies target protein from more than one gene (HPA tissue IHC). EIF3C has no transmembrane segment (UniProt Q99613: topology).
| Clear cytoplasmic chromogen in appendix or colon glandular cells, with nuclei mainly defined by counterstain. | This matches a documented high-staining cell type and the expected compartment (HPA tissue IHC: High in glandular cells; UniProt Q99613: cytoplasm). Assess the pattern in identifiable cells rather than treating overall section darkness as a positive result. |
| Predominantly nuclear, membrane-rim, or extracellular staining with little cytoplasmic signal. | That distribution conflicts with the reported cytoplasmic location (UniProt Q99613; HPA subcellular: cytosol). Review morphology, counterstain, and detection controls before interpreting it as EIF3C; unusual compartment staining alone does not establish a new location. |
| Strong staining confined to cells expected to stain low, while adjacent expected-positive cells show little signal. | Cholangiocytes, cardiomyocytes, myocytes, fibroblasts, and adipocytes are listed as Low, not absent (HPA tissue IHC). Consider off-target binding or endogenous detection activity, but first confirm cell identity and compare staining within the same run. |
| Diffuse chromogen across cells and empty tissue spaces, without a defined cytoplasmic pattern. | This is less persuasive than cell-associated cytoplasmic staining (UniProt Q99613; HPA tissue IHC). As a general IHC assessment, compare a reagent control and inspect washing, blocking, and detection background before assigning cell-level positivity. |
| No detectable cytoplasmic signal in a well-preserved appendix or colon glandular-cell section. | Those cells are reported High (HPA tissue IHC), so an all-negative run warrants a technical check. Absence on one section does not establish biological loss; inspect the antibody, retrieval and detection steps, and an independently stained positive section. |
💡Expected EIF3C appearanceCall a section positive when identifiable HPA high-staining cells show clear cytoplasmic chromogen above surrounding background—such as appendix or colon glandular cells (HPA tissue IHC: High; UniProt Q99613: cytoplasm); dominant nuclear or diffuse acellular color is a warning pattern, not sufficient evidence of EIF3C.
| Cell and tissue choice | Appendix, breast, colon, and duodenal glandular cells offer documented High staining; liver cholangiocytes and muscle cells are Low comparators, not true negatives (HPA tissue IHC). |
| Compartment and protein topology | EIF3C is cytoplasmic, with no transmembrane segment or signal peptide (UniProt Q99613); HPA ICC-IF supports cytosol. Read cytoplasmic signal in the cell context, without expecting a membrane outline. |
| Antibody evidence and specificity | The listed antibodies have Supported IHC status (HPA antibodies), while tissue reliability is Supported with medium staining–RNA agreement and a multi-gene-target caution (HPA tissue IHC). Validation supports pattern comparison, not attribution of every stained cell to EIF3C alone. |
| Isoforms and epitope | Two isoforms are listed (UniProt Q99613), but the payload gives no antibody epitope or isoform-specific staining evidence. Do not infer which isoform produced a positive IHC signal. |
| IF/ICC Q&A: what pattern should appear? | Expect cytosolic fluorescence (HPA subcellular: Cytosol, supported; UniProt Q99613: cytoplasm). HPA lists ICC-IF images in A-431, U-251MG, U2OS, Hep-G2, and MCF-7; this is a localisation reference, not an IF protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected-positive glandular cells are blank. | The run may have weak antibody or detection performance; appendix and colon glandular cells are reported High (HPA tissue IHC). | Check section integrity, antibody preparation, retrieval execution, detection reagents, and a known-positive section. Adjust conditions only after locating the failed step; no EIF3C-specific retrieval setting is supplied. |
| Color is strongest in nuclei or along cell borders. | The compartment conflicts with cytoplasmic EIF3C (UniProt Q99613; HPA subcellular: cytosol); counterstain or nonspecific signal may confuse localisation. | Inspect higher-magnification cell boundaries and a detection control. Score only convincing cell-associated cytoplasmic signal; document persistent unexpected staining separately. |
| Low-staining cell types look as dark as expected-positive cells. | HPA lists cholangiocytes, cardiomyocytes, myocytes, fibroblasts, and adipocytes as Low rather than negative (HPA tissue IHC); unexpected intensity also raises a specificity question. | Verify cell identity and compare the positive and low-staining areas on the same slide. Check reagent controls and, if available, an independently validated antibody before assigning the discrepancy to biology. |
| Background obscures cytoplasmic boundaries. | Diffuse color can arise from general chromogenic IHC background; a defined cytoplasmic pattern is expected (HPA tissue IHC; UniProt Q99613). | Compare a reagent control, then review blocking, washing, and detection conditions using standard IHC practice. Reassess signal in intact, identifiable cells after background is controlled. |
| Two IHC antibodies give different distributions. | HPA lists Supported IHC antibodies, but also reports only medium staining–RNA agreement and cautions that antibodies target protein from more than one gene (HPA antibodies; HPA tissue IHC). | Compare each antibody against the same HPA high-staining cells and expected cytoplasmic location (HPA tissue IHC; UniProt Q99613). Report the disagreement rather than combining the signals into one score. |
| A negative section is being treated as proof that EIF3C is absent. | HPA reports low tissue RNA specificity and general cytoplasmic protein expression; its Low cell categories are not negative controls (HPA tissue IHC). | Confirm that expected-positive cells stained in the same run, review morphology and controls, and describe the result as undetected under the tested conditions unless independent evidence supports absence. |
Experiment evidence
IHC & IF images showing EIF3C staining
A06470-1 · IHCIHC analysis of EIF3C using anti-EIF3C antibody (A06470-1). EIF3C was detected in a paraffin-embedded section of human lymphoma tissue. Heat
A06470-1 · IHCIHC analysis of EIF3C using anti-EIF3C antibody (A06470-1). EIF3C was detected in a paraffin-embedded section of human renal carcinoma tissu
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000184110-EIF3C/tissue/cerebral+cortex#imid_14401228
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000184110-EIF3C/tissue/colon#imid_14401146
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000184110-EIF3C/tissue/liver#imid_14401163
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000184110-EIF3C/tissue/kidney#imid_14394564
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB034045; green = EIF3C, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB034045; green = EIF3C, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for EIF3C IHC & IF
🧪Run appendix first and look for staining in glandular cells (HPA: High in appendix glandular cells). HPA detects EIF3C in all 45 scored tissues, so there is no validated negative tissue; use no-primary and isotype controls, and do not treat unstained cells within the appendix section as confirmed biological negatives (HPA: no negative rows).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: None in HPA: EIF3C 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 EIF3C in A-431, U-251MG, U2OS, Hep-G2, MCF-7, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section and a concentration-matched rabbit IgG isotype control for the rabbit primary (caption: rabbit anti-EIF3C); use EIF3C knockout material as a biological negative if available. For chromogenic staining, block endogenous peroxidase and assess endogenous biotin background because the illustrated detection uses a streptavidin–biotin complex with DAB (caption: SABC/DAB).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A06470-1 paraffin-section caption does not state a fixative (caption: fixative unreported). That caption uses heat retrieval in EDTA at pH 8.0, but it does not establish that retrieval is required for every specimen; the supplied evidence does not show that frozen sections or IF are easier (caption: EDTA pH 8.0 retrieval; HPA: cytosol supported by ICC-IF). In appendix, distinguish cellular cytoplasmic staining from background in gland lumens and inflammatory areas when scoring (HPA: High in appendix glandular cells; UniProt Q99613: cytoplasm).
HPA tissue IHC evidence for EIF3C
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: EIF3C is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced EIF3C IHC Tips
Troubleshoot EIF3C staining in paraffin sections by checking cytoplasmic localisation, retrieval conditions and antibody specificity (UniProt Q99613; HPA tissue IHC).
What retrieval conditions should I try first for weak EIF3C staining?
Start with heat mediated antigen retrieval in EDTA, pH 8.0, for the catalog antibody in paraffin sections (datasheet A06470-1). Keep the antibody incubation at the reported 2 μg/ml overnight at 4°C while assessing retrieval, so changes in staining are easier to attribute to that step (datasheet A06470-1; standard IHC practice). If staining remains weak, compare a carefully timed citrate retrieval trial against the EDTA condition on adjacent sections, checking morphology and background (standard IHC practice). The caption specifies the buffer and pH but no heating duration, so establish a reproducible heating and cooling schedule locally rather than assigning it a product specific time (datasheet A06470-1).
Could fixation explain weak or uneven EIF3C staining?
Target specific EIF3C sensitivity to fixative type or fixation duration is unknown from the supplied evidence (selected image caption A06470-1). The catalog image documents a paraffin embedded lymphoma section but does not report its fixative, so it cannot establish an EIF3C fixation requirement (selected image caption A06470-1). For troubleshooting, compare sections with documented processing histories, keep section thickness and retrieval consistent, and inspect whether staining varies with tissue preservation (standard IHC practice). Record fixation type and duration for new specimens, because these variables can alter antigen accessibility in paraffin section IHC generally; avoid assigning any observed difference to EIF3C until it is reproduced under controlled conditions (standard IHC practice).
Where should convincing EIF3C signal appear in tissue sections?
Score cytoplasmic staining as the expected pattern: EIF3C is annotated in the cytoplasm, and the tissue profile reports general cytoplasmic expression (UniProt Q99613 subcellular location; HPA tissue IHC). Its role in the eIF3 translation initiation complex supports examining cellular cytoplasm rather than treating isolated nuclear staining as confirmation (UniProt Q99613 function and subunit). Compare signal in intact cells with the local background and with a no primary control before assigning a positive call (standard IHC practice). If nuclear only or sharp membranous staining dominates, review morphology, detection background and antibody specificity; EIF3C has no annotated transmembrane segment (UniProt Q99613 topology; standard IHC practice).
Can this stain distinguish EIF3C isoforms or modified epitopes?
Do not interpret this catalog stain as isoform specific without a mapped antibody epitope and isoform validation: EIF3C has 2 annotated isoforms (UniProt Q99613 isoforms; datasheet A06470-1). The record identifies a PCI domain at residues 673–849 and modified residues including N terminal phosphoserines, but neither establishes which sequence this antibody recognises (UniProt Q99613 domains and modified residues; datasheet A06470-1). Compare the antibody's documented immunogen with isoform sequences if that information becomes available, then validate any predicted difference experimentally (standard IHC practice). A shared epitope could report both isoforms, while altered epitope accessibility after retrieval could change intensity without reflecting abundance (standard IHC practice).
How should I assess EIF3C in a multiplex IF experiment?
Treat IF as a separate application requiring its own validation; the supplied catalog image documents chromogenic IHC in a paraffin section (selected image caption A06470-1). Pair EIF3C with a marker for the cell type being assessed and inspect cytosolic overlap within segmented cells, since the supported subcellular location is cytosol (HPA subcellular; standard IF practice). Choose a fluorophore channel with low measured tissue autofluorescence and include single stain controls to evaluate bleed through (standard IF practice). Because EIF3C has no annotated transmembrane segment and is cytosolic, test gentle permeabilisation for intracellular antibody access, then check that cell morphology and marker staining remain interpretable (UniProt Q99613 topology; HPA subcellular; standard IF practice).
How can I reduce diffuse brown background without losing EIF3C signal?
First inspect no primary and detection only controls to separate antibody associated signal from chromogen or tissue background (standard IHC practice). The catalog example uses 10% goat serum blocking, a biotinylated secondary, streptavidin biotin detection and DAB; these are documented conditions for that image (selected image caption A06470-1). In a peroxidase based workflow, check peroxide blocking and consider a biotin control if background follows the detection chemistry (standard IHC practice). Titrate primary antibody around the reported 2 μg/ml, adjust blocking or washes one variable at a time, and retain interpretable cytoplasmic staining in positive cells as the endpoint (datasheet A06470-1; HPA tissue IHC; standard IHC practice).
What should I measure when comparing EIF3C across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Score cytoplasmic staining within a predefined cell population, reporting both the percentage of positive cells and intensity or a consistent H-score (HPA tissue IHC; standard IHC scoring practice). Use matched section thickness, retrieval, antibody concentration, DAB development and imaging settings across groups, because each can shift apparent staining intensity (standard IHC practice). Normalise cell counts to the number of eligible, intact cells; for spatial comparisons, report positive cell density per mm² of viable tissue and exclude necrosis (standard histology quantification practice). Document thresholds and cell type annotations before comparing groups, since HPA reports low tissue specificity and variable cell type staining (HPA tissue IHC).
How do I distinguish genuine EIF3C staining from tissue artefact?
Give greatest weight to reproducible cytoplasmic staining in intact cells, the pattern reported for EIF3C in tissue and supported for its cytosolic location (HPA tissue IHC; HPA subcellular). Compare cell identity and staining distribution with morphology: HPA reports high staining in several epithelial and other cell populations but low staining in some muscle and stromal populations (HPA tissue IHC). Be cautious with staining confined to section edges, necrotic regions, nuclei or vessel contents, and check no primary and peroxidase controls for artefact (standard IHC practice; UniProt Q99613 subcellular location). The HPA assessment is Supported yet cautions that its antibodies target protein from more than one gene, so use an independent specificity check before claiming EIF3C specific differences (HPA tissue IHC).
Boster reagents
Best EIF3C / Eukaryotic translation initiation factor 3 subunit C IHC Antibodies
The IHC-validated antibody has images from human lymphoma and renal carcinoma paraffin sections (catalog image captions). Listed reactivity covers human, mouse and rat (datasheet: reactivity).
Real IHC data
Anti-EIF3C Antibody ®
Cat # A06470-1
A06470-1 is listed for IHC, with images of human lymphoma and renal carcinoma paraffin sections (datasheet: applications; catalog image captions). Its listed reactivity is human, mouse and rat, while the supplied IHC images show human tissue (datasheet: reactivity; catalog image captions).
Which to pick: Choose A06470-1 for paraffin-section IHC: its images document EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody in human tissue (catalog image captions). No IF/ICC application or IF image is supplied for A06470-1, so an IF/ICC choice is unsupported (datasheet: applications; catalog: IF image alts). For cross-species work, A06470-1 lists human, mouse and rat reactivity, but its IHC images show human tissue only; the fixative is unreported (datasheet: reactivity; catalog image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q99613 (EIF3C_HUMAN, Eukaryotic translation initiation factor 3 subunit C).
- Human Protein Atlas. EIF3C tissue IHC expression (reliability: Supported).
- Human Protein Atlas. EIF3C subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. EIF3C antibody validation summary (4 antibodies).
- Eukaryotic translation initiation factor 3 subunit C is associated with acquired resistance to erlotinib in non-small cell lung cancer. Oncotarget 2018 — PMC6331022.
- The m6A reader YTHDF1 promotes ovarian cancer progression via augmenting EIF3C translation. Nucleic acids research 2020 — PMC7144925.
- Knockdown of EIF3C promotes human U-2OS cells apoptosis through increased CASP3/7 and Chk1/2 by upregulating SAPK/JNK. OncoTargets and therapy 2019 — PMC6389005.
- EIF3C Promotes Lung Cancer Tumorigenesis by Regulating the APP/HSPA1A/LMNB1 Axis. Disease markers 2022 — PMC9492341.
- PubMed PMID:8995409 — UniProt-cited evidence.
- PubMed PMID:10493829 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for EIF3C (Eukaryotic translation initiation factor 3 subunit C). Expected localisation General cytoplasmic staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06470-1). Validated IHC antibody: A06470-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (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 lymphoma tissue; fixative not specified (datasheet A06470-1); 2. Fixation: Image fixative and duration unreported (datasheet A06470-1); 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 A06470-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06470-1); 8. Primary antibody: Rabbit anti-EIF3C, 2-5 μg/ml (datasheet A06470-1); 9. Primary incubation: Overnight at 4 °C (datasheet A06470-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Multi-gene protein targeting may confound scoring (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unreported (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). Evidence: UniProt Q99613, HPA tissue IHC · Supported, HPA ICC-IF · cytosol., Datasheet A06470-1, PMC6331022, PMC7144925. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/EIF3C/immunohistochemistry-ihc-guide.