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EIF1AY / Eukaryotic translation initiation factor 1A, Y-chromosomal · IHC design guide
Design Immunohistochemistry for EIF1AY
Plan EIF1AY paraffin-section IHC using the catalog antibody's 1:100–1:300 dilution range (datasheet A30678). Interpret cytoplasmic and nuclear staining cautiously because HPA rates the tissue-IHC evidence uncertain and warns that the antibody targets protein from more than one gene (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
EIF1AY 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 | Tissue: cytoplasm and nuclei (HPA tissue IHC); molecular: cytoplasm (UniProt) | |
| Staining pattern | Glandular, lymphoid and hematopoietic cells: cytoplasm and nuclei (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A30678) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Skeletal muscle+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Cross-gene staining is possible; reliability is uncertain (HPA tissue IHC) | |
| Regulation | RNA enhanced in heart muscle and tongue (HPA tissue RNA) | |
| Isoform / epitope | No isoforms reported; one 1–144 chain (UniProt) |
Section 1
Recommended EIF1AY IHC & IF Protocols
The catalog antibody protocol (datasheet A30678) and one published EIF1AY IHC protocol (PMC13018312) provide starting conditions for paraffin sections.
| Sample | Paraffin-embedded Human brain tissue; fixative not specified (datasheet A30678) |
| Fixation | Image fixative and duration unreported (datasheet A30678); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 8.0 (datasheet A30678); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-EIF1AY, 1:100 - 1:300 (datasheet A30678) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | EIF1AY-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic and nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA pH 8.0 retrieval for the catalog antibody (datasheet A30678); the published protocol does not specify retrieval (PMC13018312).
Section 2
What Is the Expected EIF1AY Staining Pattern?
EIF1AY is a soluble cytoplasmic translation initiation factor with no transmembrane segment (UniProt O14602: cytoplasm; topology). In paraffin-section IHC, look for cytoplasmic staining in cell populations reported as high, including adrenal glandular cells, esophageal squamous epithelial cells, and bone-marrow hematopoietic cells (HPA: High). Interpret that pattern cautiously: HPA rates its tissue IHC Uncertain because the antibody targets proteins from more than one gene (HPA: reliability).
| Cytoplasmic staining in adrenal glandular cells, esophageal squamous epithelium, or bone-marrow hematopoietic cells (HPA: High). | This agrees with the reported tissue distribution and UniProt cytoplasmic location (HPA: High; UniProt O14602: cytoplasm). It supports a plausible staining pattern, but does not identify EIF1AY specifically: HPA rates the tissue IHC Uncertain because the antibody targets proteins from more than one gene (HPA: reliability). |
| Predominantly nuclear staining, with little or no cytoplasmic signal in a reported high-staining cell population (HPA: High). | Check for an artefact or a detection problem before calling this the expected pattern: UniProt places EIF1AY in the cytoplasm, and HPA ICC-IF places the detected signal in the cytosol (UniProt O14602; HPA: ICC-IF cytosol). Nuclear signal alone is not proof of artefact, however; HPA also describes general nuclear expression in tissue IHC (HPA: tissue profile, Uncertain). |
| Strong staining in skeletal myocytes or smooth-muscle cells (HPA: Not detected). | That conflicts with the supplied HPA tissue IHC observations and warrants a specificity check (HPA: Not detected; HPA: IHC Uncertain). Consider cross-reactivity or endogenous chromogen-producing activity as possible causes (standard IHC practice); the slide alone cannot distinguish them, and the HPA profile does not prove an EIF1AY-specific negative result. |
| Diffuse colour across tissue, empty areas, or many unrelated structures, with little cell-level distinction (standard IHC practice). | Treat this as background until controls show otherwise. Nonspecific antibody binding, incomplete blocking, or residual endogenous detection activity can obscure a cytoplasmic pattern (standard IHC practice). The supplied HPA data describe cellular patterns and validation status; they do not attribute diffuse background to a particular EIF1AY property (HPA: tissue profile). |
| No staining in an adrenal glandular-cell section that is otherwise interpretable (HPA: High). | Check whether the positive control and detection system worked before interpreting the test section (standard IHC practice). HPA reports high staining in adrenal glandular cells, but its IHC reliability is Uncertain; one negative slide cannot establish biological absence or confirm antibody specificity (HPA: High; HPA: reliability). |
💡Expected EIF1AY appearanceA plausible positive is discernible cytoplasmic chromogen in reported high-staining cell populations, such as adrenal glandular cells (UniProt O14602: cytoplasm; HPA: High); strong smooth-muscle-cell staining is suspicious because HPA reports it as Not detected, while the tissue IHC remains Uncertain (HPA: tissue profile; reliability).
| Antibody validation (HPA: HPA002561 IHC Uncertain) | HPA flags targeting of proteins from more than one gene; a matching tissue pattern cannot by itself assign the chromogen to EIF1AY (HPA: reliability; HPA002561 IHC Uncertain). |
| Compartment evidence (UniProt O14602: cytoplasm; HPA: ICC-IF cytosol) | Cytoplasmic signal fits UniProt and the approved HPA ICC-IF location. HPA tissue IHC also reports general nuclear expression, so nuclear signal needs cautious interpretation (HPA: tissue profile, Uncertain). |
| Tissue distribution (HPA: High; Low; Not detected) | HPA reports high glandular, lymphoid, hematopoietic, and esophageal epithelial staining; low glomerular-cell and cardiomyocyte staining; and no detected muscle-cell staining. These are observations from uncertain IHC, not validated EIF1AY-only controls (HPA: tissue profile; reliability). |
| Protein architecture (UniProt O14602: topology; processing) | UniProt lists a 144-residue chain, no transmembrane segment, and no signal peptide or propeptide. These support interpretation as a cellular, soluble target; they do not predict retrieval conditions or fixation sensitivity (UniProt O14602: topology; processing). |
| IF/ICC evidence (HPA: cytosol approved) | HPA's cytosol call is approved for ICC-IF, but it carries a multiple-gene-targeting caution. It helps compare compartments; it does not validate the paraffin-section IHC signal as EIF1AY-specific (HPA: subcellular summary; HPA002561 IHC Uncertain). |
| Situation | Likely cause | Next action |
|---|---|---|
| No chromogen in a reported high-staining tissue, such as adrenal glandular cells (HPA: High). | The assay may have failed, or this section may differ from the HPA observation; HPA IHC is Uncertain (standard IHC practice; HPA: reliability). | Review the positive-control section, tissue preservation, primary-antibody step, and detection reagents before scoring absence (standard IHC practice). Do not infer an EIF1AY-specific negative from one slide (HPA: reliability). |
| Nuclear signal dominates while cytoplasm is nearly blank (UniProt O14602: cytoplasm). | Detection artefact is possible, although HPA's uncertain tissue IHC also records general nuclear expression (standard IHC practice; HPA: tissue profile). | Compare the cytoplasm with a reported high-staining control and inspect negative controls (standard IHC practice; HPA: High). Record nuclear staining separately rather than automatically counting it as EIF1AY-positive (HPA: IHC Uncertain). |
| Smooth-muscle cells or skeletal myocytes stain strongly (HPA: Not detected). | Cross-reactivity or endogenous detection activity may produce discordant staining (standard IHC practice). HPA's multiple-gene warning makes target assignment especially uncertain (HPA: reliability). | Check a no-primary control for detection background and compare the tissue pattern with HPA's Not detected entries (standard IHC practice; HPA: tissue profile). Treat persistent staining as unresolved until independently validated. |
| Diffuse chromogen prevents cell-level scoring (standard IHC practice). | Nonspecific binding or endogenous activity can raise background in chromogenic IHC (standard IHC practice). The supplied sources identify no EIF1AY-specific cause (UniProt O14602; HPA: tissue profile). | Inspect no-primary controls and review blocking, washing, and detection steps (standard IHC practice). Score only distinguishable cellular staining; do not convert a uniform haze into a positive tissue call. |
| Cardiomyocytes look weaker than adrenal glandular cells (HPA: Low in cardiomyocytes; High in adrenal glandular cells). | The difference agrees with HPA's observed levels, although those levels come from uncertain tissue IHC (HPA: tissue profile; reliability). | Score intensity by cell type with comparable slide processing and a positive control (standard IHC practice). Do not require every tissue to match the strongest HPA category or infer EIF1AY abundance from chromogen alone (HPA: reliability). |
| Does approved HPA ICC-IF cytosol staining establish an IHC-P protocol or confirm EIF1AY specificity? (HPA: ICC-IF approved). | No: the ICC-IF location has a multiple-gene-targeting caution, and HPA002561 remains IHC Uncertain (HPA: subcellular summary; HPA: antibody validation). | Use the ICC-IF result only as compartment context. Evaluate paraffin-section IHC with its own controls and validation; this section supplies no IF/ICC protocol option (HPA: ICC-IF approved; HPA: IHC Uncertain). |
Experiment evidence
IHC & IF images showing EIF1AY staining
A30678 · IHCImmunohistochemical analysis of paraffin-embedded Human brain. Antibody was diluted at 1:100(4° overnight). High-pressure and temperature Tr
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000198692-EIF1AY/tissue/cerebral+cortex#imid_2495764
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000198692-EIF1AY/tissue/colon#imid_2487726
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000198692-EIF1AY/tissue/liver#imid_2487735
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000198692-EIF1AY/tissue/kidney#imid_2487720
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA002561; green = EIF1AY, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA002561; green = EIF1AY, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for EIF1AY IHC & IF
🧪Run adrenal gland first and assess staining in glandular cells (High; HPA: adrenal gland). Use skeletal muscle myocytes as the negative tissue (HPA: Not detected); the supplied HPA rows do not establish a negative cell population within adrenal gland, so judge background there against the no-primary control.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Skeletal muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show EIF1AY in A-431, U-251MG, U2OS, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) slide and an isotype control matched to the primary antibody’s host species and antibody class; confirm biological specificity with EIF1AY knockout material or an immunogen-peptide block (selected-SKU caption: peptide-block negative control). For chromogenic detection, block endogenous peroxidase and check for endogenous biotin if using biotin-based detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A30678 paraffin-section caption does not state a fixative (selected-SKU caption: fixative unreported). That caption used 1:100 antibody overnight at 4°C after high-temperature Tris-EDTA retrieval at pH 8.0; it does not establish that retrieval is required for every specimen, or that frozen sections or IF are easier (selected-SKU caption). Interpret tissue staining cautiously because the supplied HPA rows also report High glandular-cell staining in cervix and endometrium for a Y-chromosomal target (HPA: cervix and endometrium; UniProt O14602: Y-chromosomal).
HPA tissue IHC evidence for EIF1AY
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Caution, targets protein from more than one gene. Pending external verification.). 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 | Lymphoid tissue | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced EIF1AY IHC Tips
Use the paraffin-section evidence as a starting point, then evaluate compartment, controls and cross-reactivity before attributing staining to EIF1AY (caption A30678; HPA tissue IHC).
Which retrieval condition should I try first for EIF1AY IHC?
Start with heat-mediated Tris-EDTA at pH 8.0 for paraffin-section antigen retrieval (datasheet A30678). The selected human brain image used high-pressure, high-temperature retrieval, so begin with that method and compare equally processed sections when adjusting heating (caption A30678; standard IHC practice). Keep the documented 1:100 primary dilution and 4 °C overnight incubation fixed while assessing retrieval, so a signal change has a clearer cause (caption A30678; standard IHC practice). If staining remains weak, try a different heating duration or buffer only as a documented fallback, checking each condition for tissue damage and increased background (standard IHC practice).
How should I troubleshoot fixation-related loss of EIF1AY staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative, and its staining cannot establish how EIF1AY responds to fixation (caption A30678). Record the actual fixative, fixation duration and processing history for each section, then compare sections prepared under controlled conditions (standard IHC practice). Use the documented Tris-EDTA pH 8.0 retrieval and 1:100 primary dilution as starting conditions when evaluating the selected antibody (datasheet A30678; caption A30678). If staining varies with processing, assess morphology and background alongside signal; neither UniProt topology nor HPA tissue patterns demonstrate a fixation effect for this target (UniProt O14602; HPA tissue IHC).
Should EIF1AY staining be cytoplasmic or nuclear in tissue sections?
Prioritise interpretable cytoplasmic staining because EIF1AY is annotated as cytoplasmic and lacks a transmembrane segment (UniProt O14602). HPA describes general cytoplasmic and nuclear tissue staining, but rates its tissue IHC reliability uncertain because the antibodies may detect proteins from more than one gene (HPA tissue IHC). Its approved cytosol assignment also carries a multiple-gene antibody caution, so nuclear signal alone does not establish EIF1AY localisation (HPA subcellular). Compare compartments within intact cells at the same staining settings, and use appropriate negative controls before scoring either pattern as target-associated signal (standard IHC practice; HPA tissue IHC).
Could an isoform or hidden epitope explain inconsistent staining?
The supplied record lists 0 isoforms and a single 1–144 chain, so an annotated EIF1AY splice isoform does not explain differing section patterns (UniProt O14602). It also lists no signal peptide, propeptide, glycosylation sites or modified residues, and places an S1-like domain at residues 22–96 (UniProt O14602). The antibody epitope is not specified here; therefore, retrieval-dependent masking and recognition of the related EIF1AX protein remain questions to test, rather than explanations established by the record (caption A30678; UniProt O14602; HPA tissue IHC). Compare matched retrieved sections with a peptide-absorbed control, while recognising that peptide competition alone cannot prove gene specificity (caption A30678; standard IHC practice).
How can I check EIF1AY localisation by multiplex immunofluorescence?
For the separate IF/ICC workflow, pair EIF1AY staining with a marker of the cell population being assessed and inspect single-channel images before interpreting overlap (standard IF practice). HPA reports high staining in adrenal gland glandular cells, but its multiple-gene antibody warning means that pattern is a candidate for comparison, not proof of EIF1AY specificity (HPA tissue IHC). Choose spectrally separated fluorophores and assess an unstained section for tissue autofluorescence before assigning faint cytosolic signal (standard IF practice; HPA subcellular). Because EIF1AY is cytoplasmic and has no transmembrane segment, assess permeabilisation for access to an intracellular epitope; the supplied evidence establishes no antibody-specific IF fixation or permeabilisation setting (UniProt O14602; standard IF practice).
How do I separate EIF1AY signal from chromogenic background?
Examine no-primary and detection-only controls for staining produced by the chromogenic workflow, including endogenous peroxidase activity, and check reagent-specific blocking steps (standard IHC practice). Evaluate background in damaged tissue, section edges and areas with poor morphology before changing the primary antibody concentration (standard IHC practice). The selected image used 1:100 primary antibody at 4 °C overnight and shows a peptide-absorbed negative control; reproduce those comparisons when practical (caption A30678). Peptide absorption can support epitope-dependent staining, but it does not resolve the HPA warning that antibodies may detect proteins from multiple genes (caption A30678; HPA tissue IHC).
What should I quantify when comparing EIF1AY IHC sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the eligible cell population and cytoplasmic scoring compartment before examining group differences, because UniProt assigns EIF1AY to the cytoplasm (UniProt O14602; standard IHC practice). Within that population, record the percentage of positive cells and staining intensity, then calculate an H-score if graded intensity is reproducible (standard IHC practice). Normalise positive-cell counts to the number of eligible cells, or report positive-cell density per mm² of viable analysed tissue when area is the denominator (standard IHC practice). Apply identical thresholds, retrieval and chromogenic development across compared sections, and report nuclear staining separately given the uncertain, multiple-gene HPA tissue pattern (standard IHC practice; HPA tissue IHC).
When is an EIF1AY-positive IHC result convincing?
A convincing result shows reproducible staining in intact cells, principally in the expected cytoplasmic compartment, with low signal in appropriate negative controls (UniProt O14602; standard IHC practice). Treat isolated nuclear staining cautiously: HPA reports nuclear as well as cytoplasmic tissue staining but flags its IHC evidence as uncertain because antibodies may target multiple genes (HPA tissue IHC). Check whether apparent positives cluster at section edges or necrotic areas, or persist in a no-primary control consistent with endogenous enzyme or detection background (standard IHC practice). HPA also reports high staining in cervix and endometrium glandular cells, so tissue-pattern agreement alone cannot establish Y-linked EIF1AY specificity (HPA tissue IHC; UniProt O14602).
Boster reagents
Best EIF1AY / Eukaryotic translation initiation factor 1A, Y-chromosomal IHC Antibodies
The rendered anti-EIF1AY card shows paraffin-embedded human brain IHC (A30678 IHC image caption). IF/ICC applications and human, mouse, and rat reactivity are catalog-listed (catalog: A30678 applications/reactivity).
Real IHC data
Anti-EIF1AY Antibody
Cat # A30678
A30678 is the rendered SKU, with an IHC image of paraffin-embedded human brain at 1:100 after Tris-EDTA pH 8.0 retrieval (A30678 IHC image caption). IHC, IF, and ICC are listed applications, but no IF image is supplied (catalog: A30678 applications/if_image_alts).
Which to pick: For tissue IHC, choose A30678: its own image caption documents paraffin-embedded human brain and a peptide pre-absorption control; the fixative is unreported (A30678 IHC image caption). For IF/ICC, A30678 is the polyclonal, application-listed option with an IF dilution of 1:50, although no IF image is supplied (catalog: A30678 dilution_raw/applications/if_dilution/if_image_alts). Both A30678 and A14440 list human, mouse, and rat reactivity; A14440 lists IHC but has no IHC image in the payload (catalog: A30678 and A14440 reactivity/applications/ihc_image_alts).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O14602 (IF1AY_HUMAN, Eukaryotic translation initiation factor 1A, Y-chromosomal).
- Human Protein Atlas. EIF1AY tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. EIF1AY subcellular location (ICC-IF): Localized to the cytosol. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. EIF1AY antibody validation summary (1 antibodies).
- Y chromosome-linked EIF1AY deletion drives sex differences in multiple myeloma. NPJ precision oncology 2026 — PMC13018312.
- PubMed PMID:9381176 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for EIF1AY (Eukaryotic translation initiation factor 1A, Y-chromosomal). Expected localisation Tissue: cytoplasm and nuclei (HPA tissue IHC); molecular: cytoplasm (UniProt). Antigen retrieval: Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A30678). Validated IHC antibody: A30678 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Skeletal muscle. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded Human brain tissue; fixative not specified (datasheet A30678); 2. Fixation: Image fixative and duration unreported (datasheet A30678); 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: Tris-EDTA pH 8.0 (datasheet A30678); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-EIF1AY, 1:100 - 1:300 (datasheet A30678); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Cross-gene staining is possible; reliability is uncertain (HPA tissue IHC); Isoform / epitope: No isoforms reported; one 1–144 chain (UniProt); Regulation: RNA enhanced in heart muscle and tongue (HPA tissue RNA). Evidence: UniProt O14602, HPA tissue IHC · Uncertain, HPA ICC-IF · cytosol. Caution: Based on antibodies targeting proteins from…, Datasheet A30678, PMC13018312. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/EIF1AY/immunohistochemistry-ihc-guide.