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IFIT3 / Interferon-induced protein with tetratricopeptide repeats 3 · IHC design guide
Design Immunohistochemistry for IFIT3
Plan paraffin-section IFIT3 IHC around cytoplasmic staining in most tissues (HPA tissue IHC). Use kidney tubule cells as a high-staining reference and adipocytes as an undetected reference (HPA tissue IHC); the IHC-validated antibody A03920 is used at 5 μg/mL (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
IFIT3 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 most tissues (HPA tissue IHC) | |
| Staining pattern | Most tissues show cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Epididymis+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 between sections. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining has low consistency with RNA data (HPA tissue IHC) | |
| Regulation | Interferon-induced expression (UniProt) | |
| Isoform / epitope | No annotated isoforms; one 1–490 chain (UniProt) |
Section 1
Recommended IFIT3 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with the published IFIT3 chromogenic IHC protocol for ESCC tissue microarrays (PMC12847741).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A03920); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| 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-IFIT3, 5 μg/mL (datasheet A03920) |
| 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 | IFIT3-positive staining in glandular cells of epididymis (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen retrieval); the ESCC excerpt does not specify retrieval (PMC12847741).
Section 2
What Is the Expected IFIT3 Staining Pattern?
IFIT3 should appear predominantly cytoplasmic in paraffin-section IHC, including glandular cells in epididymis and seminal vesicle, kidney tubular cells, and skin melanocytes (HPA tissue IHC). Cytosolic and mitochondrial localisation is supported by ICC-IF (HPA subcellular; UniProt O14879). IFIT3 has no transmembrane segment (UniProt O14879 topology). HPA rates the tissue IHC pattern Approved, while reporting low consistency between antibody staining and RNA expression (HPA tissue IHC).
| Clear cytoplasmic staining in kidney tubular cells or epididymal glandular cells. | This matches cell types scored High by HPA and the reported cytoplasmic tissue pattern (HPA tissue IHC). Judge staining within the named cells, alongside morphology and counterstain. A section-wide positive impression can conceal unstained neighbouring cell types; it does not establish that every cell in the tissue expresses IFIT3. |
| Predominantly nuclear staining, with little or no cytoplasmic signal. | A nuclear-only result conflicts with IFIT3's reported cytoplasmic and mitochondrial locations (UniProt O14879; HPA subcellular). Treat it as suspect staining and review controls, counterstain alignment, and detection before assigning it to IFIT3. The supplied sources do not support a nuclear staining pattern as the expected IHC result. |
| Strong staining in adipocytes, skeletal-muscle myocytes, or pancreatic exocrine cells. | Those specific cell types are scored Not detected, so strong signal there is unexpected under the HPA tissue pattern (HPA tissue IHC). Consider antibody cross-reactivity or endogenous detection activity; check the omission and detection controls. Do not call the entire tissue negative when HPA identifies only a particular cell type. |
| Diffuse colour over cells, extracellular areas, and the whole section. | Broad, poorly localised deposition cannot establish the reported cytoplasmic pattern (HPA tissue IHC). Uneven blocking, washing, or detection can produce background (standard IHC practice). Compare the negative control and look for cell boundaries before scoring intensity; widespread colour alone is weak evidence for IFIT3. |
| No staining in kidney tubular cells or seminal-vesicle glandular cells. | An absent result conflicts with HPA's High staining in these named cells (HPA tissue IHC), but one negative section cannot establish biological absence. Check whether the expected cells are present and whether a positive control worked; then review the antibody, retrieval, and detection steps as general IHC workflow checks. |
💡Expected IFIT3 appearanceCall a result positive when defined cells, such as kidney tubular cells or epididymal glandular cells, show appreciable cytoplasmic staining consistent with their High HPA scores; isolated nuclear colour or uniform staining in HPA-negative cell types is suspect (HPA tissue IHC; HPA subcellular).
| Cell type and tissue context | HPA scores epididymal and seminal-vesicle glandular cells, kidney tubular cells, and melanocytes High; several other named cell types are Not detected (HPA tissue IHC). Select and score the specified cells rather than treating a tissue label as a uniform positive or negative control. |
| Compartment and optical resolution | IFIT3 is reported in cytoplasm and mitochondria (UniProt O14879; HPA subcellular). Routine chromogenic IHC supports a cytoplasmic readout; it should not be used to claim resolved mitochondrial colocalisation without a suitable method (standard IHC practice). IFIT3 has no annotated transmembrane segment (UniProt O14879 topology). |
| Strength of tissue-pattern evidence | HPA labels its tissue IHC reliability Approved but notes low consistency between antibody staining and RNA expression (HPA tissue IHC). Its listed antibody HPA059914 is IHC Approved, without an Enhanced designation in the supplied record (HPA antibodies). Interpret an unexpected pattern with independent controls rather than treating approval as proof of specificity. |
| Biological expression context | IFIT3 is interferon induced, and protein expression is reported higher in PBMCs and monocytes from SLE patients than healthy individuals (UniProt O14879). This supports context-dependent expression, but it supplies no expected staining intensity for a particular paraffin section. Record specimen context when comparing cases. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells are blank. | The named cells may be absent from the section, or an IHC workflow step may have failed; HPA reports High staining in kidney tubules and epididymal glandular cells (HPA tissue IHC). | Confirm morphology and a working positive control. Review the validated IHC procedure's retrieval, antibody incubation, and detection steps without assuming a target-specific fixation effect (standard IHC practice). |
| Signal is mainly nuclear. | The compartment conflicts with the reported cytoplasmic and mitochondrial localisation (UniProt O14879; HPA subcellular); counterstain or nonspecific signal may complicate the readout (standard IHC practice). | Compare the primary-antibody omission control, check nuclear versus cytoplasmic boundaries, and reassess the staining before scoring those nuclei as IFIT3 (standard IHC practice). |
| HPA-negative cell types stain strongly. | Adipocytes and skeletal-muscle myocytes are Not detected in the supplied HPA tissue pattern (HPA tissue IHC). Cross-reactivity or endogenous detection activity is possible, rather than established. | Check a primary-antibody omission control and the detection system's relevant endogenous-activity control; compare cell morphology with an HPA-positive cell type (standard IHC practice; HPA tissue IHC). |
| The section has uniform haze or patchy precipitate. | Background can arise from blocking, washing, or detection conditions (standard IHC practice). It obscures the cell-restricted cytoplasmic pattern described by HPA (HPA tissue IHC). | Inspect the negative control and reagent quality, then optimise blocking and washes within the validated IHC workflow; score only interpretable cellular staining (standard IHC practice). |
| Bronchial staining looks concentrated near cilia. | HPA lists Medium staining in bronchial ciliated cells at ciliary rootlets (HPA tissue IHC). This restricted observation needs cell-level interpretation. | Check that staining follows ciliated-cell morphology and compare with the control; avoid treating every apical deposit as specific IFIT3 staining (HPA tissue IHC; standard IHC practice). |
| Can IF/ICC help check the compartment? | HPA reports supported cytosolic and mitochondrial ICC-IF localisation and lists A-431, HeLa, and ASC52telo images (HPA subcellular). | Use the separate IF/ICC guide for that application; compare its compartment readout with the IHC cytoplasmic pattern. The supplied ICC-IF evidence does not define an IHC protocol option (HPA subcellular; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing IFIT3 staining
A03920 · IHCImmunohistochemistry of IFIT3 in human liver tissue with IFIT3 antibody at 5 μg/mL.
A03920-1 · IHCImmunohistochemistry of paraffin-embedded human lung cancer using IFIT3 antibody.
A03920-1 · IF
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000119917-IFIT3/tissue/cerebral+cortex#imid_16744451
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000119917-IFIT3/tissue/colon#imid_16744385
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000119917-IFIT3/tissue/liver#imid_16744386
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000119917-IFIT3/tissue/kidney#imid_16744371
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA059914; green = IFIT3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA059914; green = IFIT3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for IFIT3 IHC & IF
🧪Run kidney first: cells in the tubules should stain (HPA: High in kidney cells in tubules). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the kidney slide, unstained cells should show only background, but the supplied HPA kidney row does not identify a validated internal-negative cell type.
Positive control tissue: Epididymis (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show IFIT3 in A-431, HeLa, ASC52telo, with annotated localisation: Mitochondria (supported), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and clonality, and IFIT3 knockout material or a validated immunizing-peptide block as a biological specificity control (standard IHC practice). For kidney IHC, block endogenous peroxidase and, if using biotin-based detection, assess endogenous biotin background (standard IHC practice).
⚠️Feasibility: No supplied target-specific evidence reports an IFIT3 fixation window or fixation effect; the selected A03920 liver IHC caption does not state a fixative (selected tissue-IHC caption: fixative not stated). IFIT3-specific antigen-retrieval dependence is also unreported, so retrieval conditions need empirical optimization for paraffin IHC (standard IHC practice). Neither frozen sections nor IF/ICC is established as easier by the supplied evidence, although IF/ICC can assess the reported cytosolic and mitochondrial localization (HPA: cytosol and mitochondria supported); kidney endogenous peroxidase may complicate chromogenic background assessment (standard IHC practice).
HPA tissue IHC evidence for IFIT3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Seminal vesicle | Glandular cells | High | Protein (IHC) | HPA → |
| Skin | Melanocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced IFIT3 IHC Tips
Troubleshoot IFIT3 staining in paraffin sections by checking retrieval, compartment, cell identity and controls before interpreting chromogenic signal.
How should I retrieve IFIT3 in paraffin sections when cytoplasmic staining is weak?
Start with citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval specification). Cool sections in buffer, then compare the same staining run with a no-primary control and a section expected to show cytoplasmic signal (standard IHC practice; HPA: cytoplasmic expression in most tissues). The selected A03920 tissue image reports 5 µg/mL antibody in human liver, but its caption does not identify a retrieval method (A03920 caption). If staining remains weak, vary retrieval time on serial sections while holding antibody concentration and detection conditions constant (standard IHC practice).
Could fixation explain weak or uneven IFIT3 staining?
Target-specific fixation sensitivity is unknown: the selected tissue-IHC caption gives an antibody concentration but does not state the fixative (A03920 caption). Record the fixative, fixation duration and section thickness for each specimen, and compare sections processed together before changing antibody concentration (standard IHC practice). For paraffin sections, test retrieval on serial sections from the same block so differences in staining can be separated from differences in specimen processing (standard IHC practice). Do not infer a preferred fixative from IFIT3 phosphorylation sites or its cytoplasmic and mitochondrial locations; those annotations do not establish fixation performance (UniProt O14879: modified residues and subcellular location).
What cellular pattern should count as plausible IFIT3 staining?
Score cytoplasmic staining as the primary tissue-IHC pattern, while allowing a mitochondrial component that may appear granular (HPA tissue IHC: cytoplasmic expression in most tissues; HPA subcellular: supported cytosol and mitochondria). IFIT3 has no annotated transmembrane segment, so a sharp cell-surface rim alone needs independent confirmation before it is called IFIT3 (UniProt O14879 topology). Compare staining with cellular morphology and a no-primary control on the same specimen (standard IHC practice). Predominantly nuclear staining is discordant with the supplied localisation evidence; review counterstain overlap, pigment and detection background before assigning it to IFIT3 (UniProt O14879: cytoplasm and mitochondrion; standard IHC practice).
How do I assess an unexpected pattern when the antibody epitope is unspecified?
The supplied record lists one 1–490 chain and no annotated isoforms, but provides no antibody epitope sequence (UniProt O14879: processing and isoforms; A03920 caption). Therefore, neither a particular phosphorylation site nor a terminal region can be assumed to control staining with this antibody (UniProt O14879: phosphoserines at 203, 237 and 478; A03920 caption). Review the antibody’s documented immunogen or epitope, if available, before attributing a staining change to epitope masking (standard IHC practice). Compare serial sections under matched retrieval and detection conditions, and seek an independent antibody or orthogonal expression measurement when specificity remains uncertain (standard IHC practice).
How can IF help resolve which cells and compartments carry the IHC signal?
Use IF as a separate validation experiment and multiplex IFIT3 with a marker identifying the cell population being scored, such as kidney tubular cells when examining tubules (HPA tissue IHC: high in kidney tubular cells; standard IF practice). Choose a fluorophore channel after checking tissue autofluorescence and include single-stain controls to assess spectral overlap (standard IF practice). Because IFIT3 is reported in cytosol and mitochondria and has no transmembrane segment, permeabilise the plasma membrane to access intracellular epitopes, then optimise detergent conditions for the antibody (HPA subcellular: cytosol and mitochondria; UniProt O14879 topology; standard IF practice). Compare compartment patterns with the chromogenic section without assuming identical signal intensity across detection methods (standard IHC/IF practice).
What should I change if DAB signal appears outside plausible IFIT3-positive cells?
Run a no-primary control and inspect unstained tissue for pigment before interpreting brown deposits as antibody staining (standard chromogenic IHC practice). Check endogenous peroxidase blocking and shorten chromogen development if diffuse DAB signal persists in the control (standard chromogenic IHC practice). Evaluate background against the expected cytoplasmic pattern and against cell populations reported as undetected, such as adipocytes, without treating those populations as absolute negative controls (HPA tissue IHC: cytoplasmic profile and adipocytes not detected). If signal concentrates at section edges, folds or damaged areas, compare intact regions and a serial section before changing the biological interpretation (standard IHC practice).
How should I score IFIT3 across sections with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and anatomical region before scoring, then record cytoplasmic intensity and percentage of positive cells to calculate an H-score if intensity grading is reproducible (standard IHC practice; HPA tissue IHC: cytoplasmic profile). Alternatively, report positive cells per mm² when cell density is the relevant endpoint (standard IHC practice). Normalise counts to the number of eligible cells or sampled tissue area, and apply the same threshold, counterstain and imaging settings across sections (standard IHC practice). Report glandular, tubular and other compartments separately when present, because IFIT3 staining varies by cell type in the supplied tissue profile (HPA tissue IHC: cell-specific levels).
When is an apparent IFIT3-positive region more likely to be artefact?
Give greatest weight to intracellular cytoplasmic staining in intact, identifiable cells, with a possible mitochondrial component (HPA tissue IHC: cytoplasmic profile; HPA subcellular: supported cytosol and mitochondria). A cell-surface-only or predominantly nuclear pattern warrants specificity checks because it does not match the supplied localisation annotations (UniProt O14879: no transmembrane segment; cytoplasm and mitochondrion). Examine edges, necrotic regions, pigment and residual endogenous peroxidase activity alongside a no-primary control before scoring focal DAB deposits (standard chromogenic IHC practice). Treat unexpected cell-type staining cautiously: the tissue atlas labels its IHC reliability Approved but reports low consistency between antibody staining and RNA expression (HPA tissue IHC: reliability description).
Boster reagents
Best IFIT3 / Interferon-induced protein with tetratricopeptide repeats 3 IHC Antibodies
Two anti-IFIT3 antibodies have real human tissue IHC images: liver for A03920 (catalog image caption) and paraffin-embedded lung cancer for A03920-1 (catalog image caption). IF is listed for A03920-1 (catalog applications).
Real IHC data
Anti-IFIT3 Antibody
Cat # A03920
Real IHC data
Anti-IFIT3 Antibody
Cat # A03920-1
A03920 is listed for human IHC-P and pictured in human liver tissue at 5 μg/mL (catalog applications, reactivity, and image caption). A03920-1 is listed for human, mouse, and rat IHC and IF, with an IHC image from paraffin-embedded human lung cancer; no IF image is supplied (catalog applications, reactivity, and image captions).
Which to pick: For human paraffin-section IHC, A03920 lists IHC-P and 5 μg/mL; its image shows human liver tissue, but the caption does not report processing or fixative (catalog applications, dilution, and image caption). A03920-1 has an IHC image from paraffin-embedded human lung cancer and lists 1:100–1:200 for IHC; the caption does not report the fixative (catalog image caption and dilution). Choose A03920-1 for an IF starting point or mouse and rat work because those applications and species are listed; ICC validation and antibody clonality are unreported (catalog applications, reactivity, and clone fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O14879 (IFIT3_HUMAN, Interferon-induced protein with tetratricopeptide repeats 3).
- Human Protein Atlas. IFIT3 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. IFIT3 subcellular location (ICC-IF): Localized to the cytosol and mitochondria..
- Human Protein Atlas. IFIT3 antibody validation summary (1 antibodies).
- IFIT3 promotes lymph node metastasis by interacting with LASP1 to activate FAK-ERK signaling in esophageal squamous cell carcinoma. Cell death & disease 2025 — PMC12847741.
- Protocol of the PANCALYZE trial: a multicenter, prospective study investigating the tumor biomarkers CXCR4, SMAD4, SOX9 and IFIT3 in patients with resected pancreatic adenocarcinoma to predict the pattern of recurrence of the disease. BMC cancer 2017 — PMC5371262.
- Interferon-induced protein IFIT3 as a molecular nexus of neuroinflammation in Alzheimer's disease and HIV-associated neurocognitive disorders. Journal of neuroinflammation 2026 — PMC13059446.
- Single-cell RNA sequencing reveals age-related heterogeneity in the tumor microenvironment of breast cancer patients. Scientific reports 2025 — PMC12635151.
- PubMed PMID:9391139 — UniProt-cited evidence.
- PubMed PMID:9828129 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for IFIT3 (Interferon-induced protein with tetratricopeptide repeats 3). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A03920 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Epididymis (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A03920); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 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-IFIT3, 5 μg/mL (datasheet A03920); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent between sections. (standard IHC practice; not target-specific); Caveat: Antibody staining has low consistency with RNA data (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one 1–490 chain (UniProt); Regulation: Interferon-induced expression (UniProt). Evidence: UniProt O14879, HPA tissue IHC · Approved, HPA ICC-IF · cytosol and mitochondria., Datasheet A03920, PMC12847741. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/IFIT3/immunohistochemistry-ihc-guide.