EIF4A1 / Eukaryotic initiation factor 4A-I · IHC design guide

Design Immunohistochemistry for EIF4A1

Plan chromogenic EIF4A1 IHC around cytoplasmic staining in several tissues (HPA tissue IHC). This guide covers section handling and controls while accounting for uncertain tissue staining reliability (HPA tissue IHC) and the catalog antibody’s EIF4A1/2/3 reactivity (datasheet A03922-2).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for EIF4A1 (IHC for EIF4A1): expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC), antibody A03922-2, validated IHC image, and IHC protocol steps — Printable **EIF4A1** IHC protocol sheet — expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC), antibody A03922-2, controls and protocol steps. Open the full EIF4A1 IHC guide →

Navigate to: Protocols Staining pattern Controls Antibodies

EIF4A1 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 in several tissues (HPA tissue IHC)
	Staining pattern	Cytoplasmic in colon glands and tonsil squamous cells (HPA tissue IHC)
	Antigen retrieval	Citrate pH 6 HIER, heat-mediated (datasheet A03922-2)
	Positive control ⓘ	Bone marrow+4 more · see all
	Negative control ⓘ	Adipose tissue+4 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	A03922-2 also targets EIF4A2/3 (datasheet A03922-2)
	Regulation	Expression regulation unreported (UniProt)
	Isoform / epitope	3 isoforms; validate epitope coverage (UniProt)

Section 1

Recommended EIF4A1 IHC & IF Protocols

Compare the catalog antibody’s IHC-P protocol (datasheet A03922-2) with two published EIF4A1 chromogenic IHC workflows (PMC9441068; PMC6857661).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human colon cancer tissues; fixative not specified (datasheet A03922-2)
Fixation	Image fixative and duration unreported (datasheet A03922-2); verify before use.
Sectioning	4–5 µm sections on charged slides (standard)
Deparaffinisation	Xylene, graded ethanol series to water (standard)
Antigen retrieval	Heat retrieval: Citrate pH 6, 20 min (datasheet A03922-2)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A03922-2)
Primary antibody	Rabbit anti-EIF4A1, 0.5-1μg/ml (datasheet A03922-2)
Primary incubation	Overnight at 4 °C (datasheet A03922-2)
Detection	Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03922-2)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	EIF4A1-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control.

💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet A03922-2). The tissue microarray workflow used a different retrieval buffer at 95 °C for 30 min (PMC9441068).

Section 2

What Is the Expected EIF4A1 Staining Pattern?

EIF4A1 is expected mainly in the cytoplasm, sometimes near the nucleus; UniProt also lists the cell membrane and stress granules after arsenate or hydrogen peroxide treatment (UniProt P60842: subcellular location). In paraffin sections, look for cytoplasmic staining in the cell populations reported positive by HPA, including bone marrow hematopoietic cells and colon glandular cells (HPA: High). Interpret tissue patterns cautiously because HPA rates its tissue IHC evidence Uncertain (HPA: reliability). EIF4A1 has no transmembrane segment (UniProt P60842: topology).

What am I looking at on my slide?
Cytoplasmic chromogenic staining in bone marrow hematopoietic cells or colon glandular cells, with tissue structures still distinguishable (HPA: High in both cell populations).	This matches the reported compartment and cell populations (HPA: cytoplasmic expression; HPA: High in bone marrow and colon). Record the fraction of stained cells and intensity separately; HPA's Uncertain reliability limits how confidently staining alone establishes specificity (HPA: reliability).
Predominantly nuclear staining, or a crisp membrane outline without appreciable cytoplasmic staining (HPA: cytoplasmic tissue profile).	Treat a nuclear dominant pattern as discordant with HPA tissue IHC and its approved cytosol ICC location (HPA: tissue profile; HPA: cytosol approved). UniProt lists a cell membrane location, so membrane staining alone warrants verification before it is called an artefact (UniProt P60842: subcellular location).
Strong staining in adipocytes or adrenal glandular cells while the expected positive cell populations are weak (HPA: Not detected in those cell populations; HPA: High in bone marrow hematopoietic cells).	The distribution conflicts with the reported tissue observations (HPA: Not detected in adipocytes and adrenal glandular cells). Check the antibody and detection controls for possible cross-reactivity or endogenous detection activity; those causes cannot be distinguished from the slide alone (general IHC practice).
Diffuse chromogen covers multiple tissue compartments, including spaces outside cells, and obscures cellular boundaries (general IHC practice).	An unreadable background pattern cannot establish the cytoplasmic distribution reported for EIF4A1 (HPA: cytoplasmic tissue profile). Examine the no-primary control and review blocking, washing, and detection conditions before assigning cell-specific staining (general IHC practice).
No staining appears in bone marrow hematopoietic cells or colon glandular cells on an otherwise interpretable section (HPA: High in both cell populations).	This disagrees with HPA's observed pattern but does not by itself prove that EIF4A1 is absent (HPA: High; HPA: reliability Uncertain). Check section quality, antigen retrieval, primary-antibody use, and detection with appropriate controls (general IHC practice).

💡Expected EIF4A1 appearanceCall a result provisionally positive when a defined cell population shows cytoplasmic staining, especially bone marrow hematopoietic or colon glandular cells reported High by HPA; isolated nuclear staining, broad extracellular color, or signal confined to a no-primary control is suspect (HPA: cytoplasmic profile; HPA: High; general IHC practice).

How each factor affects the staining
Tissue and cell choice (HPA: tissue IHC)	HPA reports High staining in bone marrow hematopoietic cells, colon, rectum, and stomach glandular cells, placental trophoblastic cells, and tonsil squamous epithelial cells (HPA: High). Its profile has Uncertain reliability, so these are observed patterns rather than guaranteed controls (HPA: reliability).
Antibody validation (HPA: antibody records)	The listed IHC antibody CAB011689 has an Uncertain IHC assessment; HPA068286 has Approved ICC status but no listed IHC status (HPA: antibody records). Do not transfer the ICC assessment to paraffin tissue staining; judge each IHC result alongside its controls (general IHC practice).
Epitope coverage and isoforms (UniProt P60842: isoforms)	UniProt lists 3 EIF4A1 isoforms and helicase regions at residues 63–234 and 245–406 (UniProt P60842: isoforms, domains). The supplied record gives no antibody epitope, so it cannot establish which isoforms an antibody detects.
Processing and topology (UniProt P60842: processing, topology)	UniProt lists no signal peptide, propeptide, or transmembrane segment and annotates a chain spanning residues 2–406 (UniProt P60842: processing, topology). These annotations support interpreting a cytoplasmic pattern; they provide no target-specific evidence about fixation or retrieval sensitivity.
IF/ICC comparison (HPA: subcellular ICC-IF)	Q: Should an IF/ICC image settle an ambiguous IHC result? A: HPA approves cytosol as the main ICC location, but cautions that its summary draws on antibodies targeting proteins from multiple genes (HPA: subcellular ICC-IF). Use it as compartment context, not IHC validation.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
The expected positive tissue shows no chromogenic signal (HPA: High in bone marrow hematopoietic cells or colon glandular cells).	Possible section, retrieval, primary-antibody, or detection failure; a negative slide alone cannot identify which step failed (general IHC practice).	Check tissue preservation and morphology, confirm the run's positive control, and review the established antigen retrieval and detection steps before interpreting absence (general IHC practice).
Both tissue and no-primary control develop color (general IHC practice).	The shared signal implicates the detection system or endogenous tissue activity rather than proving specific EIF4A1 binding (general IHC practice).	Review the blocking step appropriate to the detection chemistry, reagent exposure, and washing; compare controls after correction (general IHC practice).
Staining is diffuse and cellular boundaries cannot be scored (general IHC practice).	Excess background can arise from antibody concentration, incomplete blocking, or inadequate washing; the appearance alone does not separate these possibilities (general IHC practice).	Adjust one workflow variable at a time using the antibody's documented IHC conditions, and retain a no-primary control (general IHC practice).
A nuclear dominant pattern replaces cytoplasmic staining (HPA: cytoplasmic tissue profile).	The compartment is discordant with HPA's tissue profile and approved cytosol ICC location; specificity remains uncertain (HPA: tissue profile; HPA: cytosol approved; HPA: reliability).	Inspect counterstain and tissue morphology, compare a reported positive cell population, and repeat with an independently validated IHC antibody if available (general IHC practice).
Adipocytes stain strongly but reported positive cells do not (HPA: Not detected in adipocytes; HPA: High in bone marrow hematopoietic cells).	The cell distribution conflicts with HPA observations; cross-reactivity, background, and sample differences remain possibilities (HPA: tissue IHC; general IHC practice).	Score each cell population separately, inspect no-primary and tissue controls, and avoid calling the adipocyte signal specific without independent support (general IHC practice).
IHC and ICC images suggest different compartments (HPA: tissue IHC; HPA: subcellular ICC-IF).	The assays use different sample preparations, and HPA's ICC summary carries a multi-gene antibody caution; the IHC tissue profile is also Uncertain (HPA: subcellular ICC-IF; HPA: reliability).	Interpret the paraffin section against its own tissue and detection controls. Use the ICC cytosol result as context, without treating it as proof of IHC antibody performance (general IHC practice; HPA: cytosol approved).

Source: UniProt P60842 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing EIF4A1 staining

IHC analysis of EIF4A using anti-EIF4A antibody (A03922-2). EIF4A was detected in paraffin-embedded section of human colon cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-EIF4A Antibody (A03922-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

A03922-2 · IHCIHC analysis of EIF4A using anti-EIF4A antibody (A03922-2). EIF4A was detected in paraffin-embedded section of human colon cancer tissues. H

IHC analysis of EIF4A using anti-EIF4A antibody (A03922-2). EIF4A was detected in paraffin-embedded section of mouse intestine tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-EIF4A Antibody (A03922-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

A03922-2 · IHCIHC analysis of EIF4A using anti-EIF4A antibody (A03922-2). EIF4A was detected in paraffin-embedded section of mouse intestine tissues. Heat

IF analysis of EIF4A using anti-EIF4A antibody (A03922-2). EIF4A was detected in immunocytochemical section of U20S cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-EIF4A Antibody (A03922-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

A03922-2 · IFIF analysis of EIF4A using anti-EIF4A antibody (A03922-2). EIF4A was detected in immunocytochemical section of U20S cell. Enzyme antigen ret

IHC analysis of EIF4A1 using anti-EIF4A1 antibody (M03922-2). EIF4A1 was detected in a paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml mouse anti-EIF4A1 Antibody (M03922-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

M03922-2 · IHCIHC analysis of EIF4A1 using anti-EIF4A1 antibody (M03922-2). EIF4A1 was detected in a paraffin-embedded section of human placenta tissue. H

IHC analysis of EIF4A1 using anti-EIF4A1 antibody (M03922-2). EIF4A1 was detected in a paraffin-embedded section of human colonic adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml mouse anti-EIF4A1 Antibody (M03922-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

M03922-2 · IHCIHC analysis of EIF4A1 using anti-EIF4A1 antibody (M03922-2). EIF4A1 was detected in a paraffin-embedded section of human colonic adenocarci

IF analysis of EIF4A1 using anti-EIF4A1 antibody (M03922-2). EIF4A1 was detected in an immunocytochemical section of HEP3B cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL mouse anti-EIF4A1 Antibody (M03922-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

M03922-2 · IFIF analysis of EIF4A1 using anti-EIF4A1 antibody (M03922-2). EIF4A1 was detected in an immunocytochemical section of HEP3B cells. Enzyme ant

IHC analysis of EIF4A1 using anti-EIF4A1 antibody (A03922-3). EIF4A1 was detected in paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-EIF4A1 Antibody (A03922-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

A03922-3 · IHCIHC analysis of EIF4A1 using anti-EIF4A1 antibody (A03922-3). EIF4A1 was detected in paraffin-embedded section of human tonsil tissue. Heat

IHC analysis of EIF4A1 using anti-EIF4A1 antibody (A03922-3). EIF4A1 was detected in paraffin-embedded section of human Lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-EIF4A1 Antibody (A03922-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

A03922-3 · IHCIHC analysis of EIF4A1 using anti-EIF4A1 antibody (A03922-3). EIF4A1 was detected in paraffin-embedded section of human Lung cancer tissue.

IF analysis of EIF4A1 using anti-EIF4A1 antibody (A03922-3). EIF4A1 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-EIF4A1 Antibody (A03922-3) overnight at 4°C. DyLight®488 conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

A03922-3 · IFIF analysis of EIF4A1 using anti-EIF4A1 antibody (A03922-3). EIF4A1 was detected in immunocytochemical section of A431 cells. Enzyme antigen

IHC analysis of EIF4A1 using anti-EIF4A1 antibody (M03922-1). EIF4A1 was detected in paraffin-embedded section of human gastric cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-EIF4A1 Antibody (M03922-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

M03922-1 · IHCIHC analysis of EIF4A1 using anti-EIF4A1 antibody (M03922-1). EIF4A1 was detected in paraffin-embedded section of human gastric cancer tissu

IHC analysis of EIF4A1 using anti-EIF4A1 antibody (M03922-1). EIF4A1 was detected in paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-EIF4A1 Antibody (M03922-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

M03922-1 · IHCIHC analysis of EIF4A1 using anti-EIF4A1 antibody (M03922-1). EIF4A1 was detected in paraffin-embedded section of human breast cancer tissue

IF analysis of EIF4A1 using anti-EIF4A1 antibody (M03922-1). EIF4A1 was detected in immunocytochemical section of CACO-2 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti-EIF4A1 Antibody (M03922-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

M03922-1 · IFIF analysis of EIF4A1 using anti-EIF4A1 antibody (M03922-1). EIF4A1 was detected in immunocytochemical section of CACO-2 cells. Enzyme antig

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000161960-EIF4A1/tissue/cerebral+cortex#imid_3643584

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000161960-EIF4A1/tissue/colon#imid_3643510

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000161960-EIF4A1/tissue/liver#imid_3643527

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000161960-EIF4A1/tissue/kidney#imid_3643528

HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA068286; green = EIF4A1, blue = nucleus, red = microtubules.

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Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for EIF4A1 IHC & IF

🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipocytes); the supplied colon data do not identify a validated internal negative cell population, so do not assume that adjacent cells should be unstained (HPA: colon glandular-cell row only).

Positive control tissue: Bone marrow (Hematopoietic cells, HPA High)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show EIF4A1 in CACO-2, U-251MG, U2OS, with annotated localisation: Cytosol (approved) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only slide and an isotype control using rabbit IgG matched to the primary antibody’s clonality and concentration (selected-SKU caption: rabbit primary antibody). Use EIF4A1 knockout material as a biological negative; block endogenous peroxidase and assess endogenous biotin background in colon sections because the stated detection uses biotin and DAB (selected-SKU caption: biotinylated secondary, SABC/DAB).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A03922-2 paraffin-section caption does not state the fixative (selected-SKU caption: fixative not stated). That caption uses citrate retrieval at pH 6 for 20 minutes, but does not establish whether retrieval is required (selected-SKU caption: citrate retrieval). Frozen sections are not shown to be easier; IF/ICC images support a cytosolic signal, while colon fluorescence may require an autofluorescence control (HPA: Cytosol approved; standard IF practice).

HPA tissue IHC evidence for EIF4A1

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. 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
Bone marrow	Hematopoietic cells	High	Protein (IHC)	HPA →
Colon	Glandular cells	High	Protein (IHC)	HPA →
Placenta	Trophoblastic cells	High	Protein (IHC)	HPA →
Rectum	Glandular cells	High	Protein (IHC)	HPA →
Stomach	Glandular cells	High	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →
Adrenal gland	Glandular cells	Not detected	Protein (IHC)	HPA →
Caudate	Glial cells	Not detected	Protein (IHC)	HPA →
Cerebellum	Cells in granular layer	Not detected	Protein (IHC)	HPA →
Cerebral cortex	Endothelial cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced EIF4A1 IHC Tips

Use the catalog antibody’s paraffin-section result as the starting point, then check staining against EIF4A1 localisation and tissue controls.

What retrieval should I try when EIF4A1 staining is weak in paraffin sections?

Begin with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A03922-2). The catalog antibody produced chromogenic staining in paraffin-embedded human colon cancer sections after this treatment, although the caption does not state the fixative (datasheet A03922-2). If staining remains weak, compare a modestly longer heat exposure on matched sections while keeping antibody concentration and detection constant (standard IHC practice). Check that the heated sections remain attached and retain interpretable morphology, because tissue loss can resemble weak antigen detection (standard IHC practice). Include a known staining control in each retrieval comparison, such as colon glandular cells reported as high by tissue IHC (HPA: High in colon glandular cells).

Could fixation explain inconsistent EIF4A1 staining between paraffin blocks?

Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not report a fixative, and its staining result cannot establish tolerance to any particular fixation condition (datasheet A03922-2). Compare blocks with documented processing histories using the same citrate pH 6, 20-minute retrieval and antibody conditions (datasheet A03922-2; standard IHC practice). Inspect morphology and tissue retention alongside staining, since differences in processing can complicate comparisons between sections (standard IHC practice). Run comparable control tissue with each batch rather than interpreting a blank study section alone (standard IHC practice). Do not infer a fixation effect from EIF4A1 topology, modifications, or tissue staining patterns (UniProt P60842; HPA tissue IHC).

Which compartment should show convincing EIF4A1 staining?

Prioritise cytoplasmic staining: EIF4A1 is annotated in cytoplasm and the perinuclear region, and tissue IHC reports cytoplasmic expression across several tissues (UniProt P60842 subcellular location; HPA tissue IHC). High staining in colon glandular cells provides a relevant cellular comparison for the selected colon cancer section (HPA: High in colon glandular cells; datasheet A03922-2). Record diffuse cytoplasmic and perinuclear signal separately from sharply nuclear or exclusively membrane-associated deposits when reviewing DAB sections (UniProt P60842 subcellular location; standard IHC practice). EIF4A1 is also annotated at the cell membrane and in stress granules, so a focal pattern deserves investigation rather than automatic rejection (UniProt P60842 subcellular location). HPA’s cytosol assignment carries a warning that its antibodies target proteins from multiple genes (HPA subcellular).

Can this antibody distinguish EIF4A1 isoforms or modification-dependent staining?

Do not assign a paraffin-section signal to one EIF4A1 isoform without epitope or isoform-specific validation: the record lists 3 isoforms, while the selected caption supplies no such specificity test (UniProt P60842 isoforms; datasheet A03922-2). EIF4A1 contains an ATP-binding helicase region at residues 63–234 and a C-terminal helicase region at 245–406 (UniProt P60842 domains). Reported modifications include acetylation and phosphorylation, but the caption does not show whether they affect this antibody’s staining (UniProt P60842 modified residues; datasheet A03922-2). If a biological comparison depends on an isoform or modified residue, first establish antibody specificity with an appropriate independent assay or validated control (standard IHC practice). Keep retrieval and detection identical across sections during that assessment (standard IHC practice).

How should I check an EIF4A1 signal in multiplex IF?

Treat IF as a separate application requiring its own antibody validation; the selected product evidence describes chromogenic staining of paraffin sections (datasheet A03922-2). Pair EIF4A1 with a validated marker for the cell population under study and assess whether its signal lies mainly in that population’s cytoplasm (standard IF practice; UniProt P60842 subcellular location). Choose spectrally separated fluorophores after checking unstained tissue autofluorescence and single-stain controls (standard IF practice). Because EIF4A1 has no transmembrane segment and its expected signal is intracellular, assess permeabilisation for access to the epitope; the epitope itself is unspecified (UniProt P60842 topology and subcellular location; standard IF practice). HPA reports cytosolic IF localisation but cautions that the antibodies used target proteins from multiple genes (HPA subcellular).

What should I investigate when DAB background obscures EIF4A1?

Start by comparing a no-primary control with the stained section to identify detection-system or tissue background (standard IHC practice). The selected example used 10% goat serum blocking, 1 μg/ml primary antibody overnight at 4°C, and a biotin-based DAB detection system (datasheet A03922-2). For that system, evaluate endogenous peroxidase blocking and, where relevant, endogenous biotin interference as general chromogenic IHC checks (standard IHC practice). If the no-primary control is clean, compare lower primary exposure or stronger washing against preservation of the expected cytoplasmic signal (standard IHC practice; UniProt P60842 subcellular location). Examine pigment, folds, and section edges separately before calling diffuse brown material EIF4A1 (standard IHC practice).

How can I score EIF4A1 IHC consistently across samples? ⚠ ANSWER MARKED FOR VERIFICATION

Define the analysed cell compartment and score cytoplasmic staining in viable cells, keeping perinuclear staining within the same predefined rule (UniProt P60842 subcellular location; standard IHC practice). An H-score combines the percentages of cells at intensity grades 0–3 and ranges from 0–300; alternatively, report percentage positive cells or positive area per mm² (standard IHC practice). Normalise counts or stained area to the number or area of eligible cells, excluding blank tissue and necrosis by a fixed rule (standard IHC practice). Hold retrieval, DAB development, image acquisition, and positivity thresholds constant across the comparison (standard IHC practice). Interpret modest differences cautiously because HPA rates its EIF4A1 tissue staining reliability as uncertain (HPA tissue IHC).

How do I distinguish true EIF4A1 staining from a tissue artefact?

A credible result follows the expected predominantly cytoplasmic or perinuclear distribution and appears in intact cells rather than only at section edges (UniProt P60842 subcellular location; standard IHC practice). Compare cell identity with reported tissue patterns: HPA lists high staining in colon glandular cells but no detection in adipocytes, while rating overall tissue IHC reliability uncertain (HPA tissue IHC). Investigate exclusively nuclear deposits, necrotic regions, folds, and edge-restricted DAB before assigning biological meaning (standard IHC practice; UniProt P60842 subcellular location). Use a no-primary control to assess endogenous enzyme or detection background, especially when the signal is diffuse (standard IHC practice). Confirm an unexpected cell distribution with an independent specificity control before treating it as EIF4A1 expression (standard IHC practice).

Boster reagents

Best EIF4A1 / Eukaryotic initiation factor 4A-I IHC Antibodies

Four catalog antibodies have IHC images from human paraffin sections (IHC captions); A03922-3 also has IF/ICC data from human A431 cells (A03922-3 IF caption).

Real IHC data

Anti-EIF4A1/2/3 Antibody ®

Cat # A03922-2

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Real IHC data

Anti-EIF4A1 Antibody ® (monoclonal, 3F11)

Cat # M03922-2

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Real IHC data

Anti-EIF4A1 Antibody ®

Cat # A03922-3

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Real IHC data

Anti-EIF4A1 Antibody ® (monoclonal, 11B8)

Cat # M03922-1

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A03922-2 is shown on human colon cancer, and M03922-2 on human placenta (respective IHC captions). A03922-3 is shown on human tonsil, and M03922-1 on human gastric cancer (respective IHC captions).

Which to pick: For EIF4A1 tissue IHC, choose A03922-3: its paraffin human tonsil image used EDTA pH 8.0 retrieval (A03922-3 IHC caption); the fixative is unreported (A03922-3 IHC caption). For IF/ICC, A03922-3 has an image from human A431 cells at 2 μg/ml (A03922-3 IF caption). For cross-species tissue work, M03922-2 is a mouse monoclonal with human, mouse and rat IHC reactivity listed (M03922-2 catalog); its IHC images show human placenta and mouse colon, with no rat tissue image listed (M03922-2 IHC captions).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry P60842 (IF4A1_HUMAN, Eukaryotic initiation factor 4A-I).
Human Protein Atlas. EIF4A1 tissue IHC expression (reliability: Uncertain).
Human Protein Atlas. EIF4A1 subcellular location (ICC-IF): Localized to the cytosol. Caution: Based on antibodies targeting proteins from multiple genes..
Human Protein Atlas. EIF4A1 antibody validation summary (2 antibodies).
Novel eIF4A1 inhibitors with anti-tumor activity in lymphoma. Molecular medicine (Cambridge, Mass.) 2022 — PMC9441068.
Aberrant Expression Of PDCD4/eIF4A1 Signal Predicts Postoperative Recurrence For Early-Stage Oral Squamous Cell Carcinoma. Cancer management and research 2019 — PMC6857661.
Expression pattern and prognostic value of key regulators for N7-methylguanosine RNA modification in prostate cancer. Acta biochimica et biophysica Sinica 2023 — PMC10195147.
Identification and Verification of m(7)G Modification Patterns and Characterization of Tumor Microenvironment Infiltration via Multi-Omics Analysis in Clear Cell Renal Cell Carcinoma. Frontiers in immunology 2022 — PMC9113293.
PubMed PMID:8493113 — UniProt-cited evidence.
PubMed PMID:14702039 — UniProt-cited evidence.
PubMed PMID:16625196 — UniProt-cited evidence.