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IFITM1 / Interferon-induced transmembrane protein 1 · IHC design guide
Design Immunohistochemistry for IFITM1
Plan IFITM1 chromogenic IHC on paraffin sections with the catalog antibody at 0.5–1 μg/mL (datasheet). Assess membranous staining in epididymal glands and account for endothelial and lymphoid signal when scoring (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
IFITM1 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 | Membranous staining in epididymal glands (HPA tissue IHC) | |
| Staining pattern | Distinct epididymal gland membranes; endothelial and lymphoid signal (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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PA1112) | |
| Caveat | Endothelial and lymphoid positivity can complicate scoring (HPA tissue IHC) | |
| Regulation | Interferon induced expression (UniProt) | |
| Isoform / epitope | No annotated isoforms; cytoplasmic vs extracellular epitope matters (UniProt) |
Section 1
Recommended IFITM1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by three published IFITM1 staining protocols (PMC4867989; PMC7404048; PMC6172064).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet PA1112); 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-IFITM1, 0.5-1μg/ml (datasheet PA1112) |
| 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 | IFITM1-positive staining in glandular cells of epididymis (HPA tissue IHC: High). HPA tissue profile: Distinct membranous expression in epididymis, also expressed in endothelial cells and in lymphoid tissue. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval); the oral cancer study used trypsin retrieval (PMC6172064).
Section 2
What Is the Expected IFITM1 Staining Pattern?
IFITM1 is a membrane protein with a transmembrane segment at residues 87–107 and cell membrane and lysosome membrane annotations (UniProt P13164 topology; UniProt P13164 subcellular). In paraffin-section IHC, expect the clearest membranous staining in epididymal glandular cells, with staining also reported in endothelial and lymphoid cells (HPA: High in epididymal glandular cells; HPA: tissue IHC profile). HPA rates its tissue IHC evidence Enhanced, citing agreement between staining and RNA expression (HPA: tissue IHC reliability).
| Crisp membrane staining in epididymal glandular cells, with little staining in adjacent unstained areas. | This matches HPA’s distinct membranous profile and High glandular-cell staining in epididymis (HPA: tissue IHC profile; HPA: High in epididymal glandular cells). Compare cells within the same section when judging whether the signal follows the expected anatomy (general IHC practice). |
| Staining follows endothelial cells or lymphoid areas outside the epididymis. | These can be expected positive populations: HPA reports endothelial staining, Medium lymphoid-tissue staining in appendix, and Medium germinal-center staining in lymph node (HPA: tissue IHC profile; HPA: appendix and lymph-node IHC). Judge the labeled cell population, rather than treating every cell in a positive tissue as positive. |
| Strong, widespread nuclear staining replaces a discernible membrane pattern. | A dominant nuclear pattern does not match the reported cell membrane or lysosome membrane locations, or HPA’s membranous tissue profile (UniProt P13164 subcellular; HPA: tissue IHC profile). Treat it as suspect and check controls before assigning it to IFITM1 (general IHC practice). |
| Signal appears mainly in a cell population HPA reports as undetected, or as uniform staining across unrelated cells. | For example, HPA reports IFITM1 as Not detected in adipocytes and cardiomyocytes (HPA: adipose-tissue and heart-muscle IHC). Consider antibody cross-reactivity or endogenous detection activity, especially if the pattern persists in a no-primary control (general IHC practice). |
| A diffuse haze obscures cell borders, or no staining appears in an epididymal positive control. | Haze prevents a reliable membrane call against HPA’s distinct membranous reference pattern (HPA: tissue IHC profile). Absence in a known-positive cell population makes a negative test section inconclusive; review the run controls and detection workflow before interpreting it (HPA: High in epididymal glandular cells; general IHC practice). |
💡Expected IFITM1 appearanceCall the IHC result positive when epididymal glandular cells show distinct, strong membrane staining, consistent with HPA’s High result; widespread nuclear color or diffuse haze without defined cells is suspect (HPA: epididymis IHC and tissue IHC profile; UniProt P13164 subcellular).
| Membrane location and topology | UniProt places IFITM1 at the cell and lysosome membranes and maps its transmembrane segment to 87–107 (UniProt P13164 subcellular and topology). The expected tissue IHC readout is membranous; the supplied topology does not identify the catalog antibody’s epitope or establish an antigen-retrieval requirement (HPA: tissue IHC profile). |
| Tissue and cell selection | The strongest listed normal-tissue example is epididymal glandular cells (High), while several endothelial, lymphoid, stromal, and epithelial populations are Medium or Low (HPA: tissue IHC). Select and score the documented cell population; a whole-section average can obscure a localized positive pattern. |
| Antibody-validation scope | HPA lists IHC as Enhanced for HPA004810 and CAB017615, and describes tissue-level agreement with RNA expression (HPA: antibody validation; HPA: tissue IHC reliability). This supports the reported tissue pattern; it does not establish the performance of an unlisted antibody or every staining condition. |
| Tumor versus normal tissue | UniProt reports greatly elevated IFITM1 levels in colon, cervical, esophageal, and ovarian cancers (UniProt P13164 tissue specificity). Do not transfer that statement to normal tissue: HPA lists Low staining in normal colon endothelial cells and normal cervical glandular cells (HPA: tissue IHC). Interpret the specimen and cell population separately. |
| Subcellular evidence by application | HPA ICC-IF supports plasma-membrane localization and labels Golgi localization uncertain; UniProt also annotates lysosome membrane (HPA: subcellular ICC-IF; UniProt P13164 subcellular). These sources make intracellular signal worth examining, but they do not establish a routine Golgi or lysosomal pattern in paraffin-section IHC. |
| Situation | Likely cause | Next action |
|---|---|---|
| Epididymal positive-control cells are blank. | The run may have failed, or the chosen conditions may not reveal detectable staining; HPA reports these cells as High (HPA: epididymis IHC). | Check the control section, primary-antibody step, detection reagents, and staining run. Review antigen retrieval as a general IHC variable; target-specific retrieval or fixation sensitivity is unreported in the supplied sources (general IHC practice). |
| Most of the section has diffuse chromogen haze. | Nonspecific binding, excess detection signal, or endogenous detection activity can obscure cell borders (general IHC practice). | Inspect the no-primary control and adjust blocking, washing, and detection conditions as appropriate (general IHC practice). Reassess for defined membrane staining in the HPA-reported positive cells (HPA: tissue IHC profile). |
| Strong nuclear color dominates the positive tissue. | This conflicts with the supplied membrane locations and HPA’s membranous tissue profile (UniProt P13164 subcellular; HPA: tissue IHC profile). | Check no-primary and other run controls, then confirm whether any cell-border staining remains (general IHC practice). Do not score isolated nuclear color as the expected IFITM1 pattern. |
| Cells reported as undetected show strong color. | Cross-reactivity or endogenous detection activity is possible; HPA reports adipocytes and cardiomyocytes as Not detected (HPA: adipose-tissue and heart-muscle IHC). | Compare the same cell type with the HPA reference pattern and examine a no-primary control (HPA: tissue IHC; general IHC practice). If controls are clean, confirm the finding independently before interpreting it as IFITM1. |
| A lymphoid or endothelial region looks weaker than epididymis. | HPA reports Medium staining in appendix lymphoid tissue, lymph-node germinal-center cells, and skin endothelial cells, versus High epididymal glandular staining (HPA: tissue IHC). | Score the documented cell population and its expected level. A weaker, localized positive can be consistent with the HPA pattern; compare it with run controls before changing conditions. |
| Q: Does an ICC-IF Golgi signal establish the expected IHC pattern? | HPA calls ICC-IF Golgi localization uncertain, while supporting plasma-membrane localization (HPA: subcellular ICC-IF). | A: No. Interpret paraffin-section staining against HPA’s membranous tissue IHC profile and its named positive cell populations (HPA: tissue IHC). Evaluate ICC-IF localization on its own guide page. |
Experiment evidence
IHC & IF images showing IFITM1 staining
PA1112 · IHCAnti-IFITM1 antibody, PA1112, IHC(P) IHC(P): Human Intestinal Cancer Tissue
A02633 · IFImmunocytochemistry of IFITM1 in Jurkat cells with IFITM1 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000185885-IFITM1/tissue/cerebral+cortex#imid_1626276
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000185885-IFITM1/tissue/colon#imid_1626218
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000185885-IFITM1/tissue/liver#imid_1626211
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000185885-IFITM1/tissue/kidney#imid_1626220
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA004810; green = IFITM1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA004810; green = IFITM1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for IFITM1 IHC & IF
🧪Run epididymis first; glandular cells should stain (HPA: High in epididymis glandular cells). Use adipose tissue as the negative, with adipocytes at background (HPA: Not detected in adipocytes); on the positive slide, assess morphologically distinct non-glandular cells as an internal background reference, without assuming every such cell is negative.
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 IFITM1 in A-431, K-562, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Golgi apparatus (uncertain), Plasma membrane (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; use host-species-matched IgG for a polyclonal antibody or subclass-matched isotype for a monoclonal antibody, and confirm specificity with an IFITM1 knockout sample or immunizing-peptide competition where available (standard IHC/IF practice). For chromogenic IHC, block endogenous peroxidase and check for endogenous biotin if using biotin-based detection; for IF, check tissue autofluorescence (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected PA1112 paraffin-section caption does not state a fixative (selected-SKU IHC(P) caption). Retrieval dependency is unreported; optimize antigen retrieval for paraffin IHC using the catalog antibody’s IHC-P protocol as the starting point (standard IHC practice). The supplied evidence does not establish that frozen sections or IF/ICC are easier, or identify an epididymis-specific artefact; IF/ICC images support plasma-membrane localization, while Golgi localization is uncertain (HPA subcellular).
HPA tissue IHC evidence for IFITM1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 → |
| Appendix | Lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Medium | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (tip of cilia) | Medium | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | Medium | 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 → |
| Bronchus | Basal cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced IFITM1 IHC Tips
Use membrane pattern, cell identity and matched controls to troubleshoot IFITM1 staining in paraffin sections; interpret fluorescence separately.
How should I adjust retrieval when IFITM1 staining is weak or uneven?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval specification). Run adjacent sections through the same cooling, antibody incubation and chromogenic detection steps so retrieval is the variable being assessed (standard IHC practice). IFITM1 is found at the cell and lysosome membranes, so assess whether staining resolves into a plausible cellular pattern rather than judging intensity alone (UniProt P13164 localisation). If staining remains weak, compare a modestly longer retrieval time on matched sections and check tissue preservation, background and a known positive control before adopting the change (standard IHC practice; HPA: high in epididymal glandular cells).
Could fixation explain weak IFITM1 staining in my paraffin sections?
Target-specific sensitivity of IFITM1 to fixative type or fixation duration is unknown from the supplied evidence (selected PA1112 caption: fixative not stated). Record the fixative, duration and processing conditions for each specimen, then compare sections processed together before attributing an intensity difference to biology (standard IHC practice). The PA1112 image identifies IHC on human intestinal cancer tissue but supplies no fixation condition to reproduce (selected PA1112 caption). If fixation varies between groups, use matched controls and examine morphology alongside staining; a retrieval change may improve signal, but it cannot establish that fixation caused the original difference (standard IHC practice).
Where should IFITM1 staining appear, and when is cytoplasmic signal suspicious?
Prioritise staining at cell boundaries, consistent with IFITM1 at the cell membrane and supported plasma membrane localisation (UniProt P13164 localisation; HPA subcellular: plasma membrane supported). Intracellular puncta can also be plausible because UniProt places IFITM1 at lysosome membranes; a Golgi assignment is listed as uncertain by HPA (UniProt P13164 localisation; HPA subcellular: Golgi uncertain). Compare the suspected pattern with morphology and a matched negative control, since diffuse chromogen across damaged tissue does not establish a membrane signal (standard IHC practice). Interpret each cell population separately: HPA reports distinct membranous staining in epididymis and expression in endothelial cells and lymphoid tissue (HPA tissue IHC profile).
How does epitope position affect interpretation of IFITM1 IHC staining?
Confirm the catalog antibody's stated immunogen or epitope before troubleshooting accessibility; its epitope is not provided in the supplied payload (supplied antibody evidence: epitope unspecified). UniProt describes a 125 amino acid chain, with a transmembrane segment at residues 87–107 and an extracellular region at 108–125 (UniProt P13164 topology). The annotated cytoplasmic regions are residues 1–36 and 58–86, so epitope position could affect accessibility after processing and retrieval (UniProt P13164 topology; standard IHC practice). No alternate isoforms or glycosylation sites are annotated; phosphoserine at residue 16 is noted, but its effect on this antibody's staining is unknown (UniProt P13164 isoforms, glycosylation and modified residues).
How can I assess IFITM1 by multiplex IF alongside the IHC findings?
Treat IF/ICC as a separate assay: the PA1112 caption documents tissue IHC and supplies no IF validation (selected PA1112 caption). Pair IFITM1 with a marker that identifies the cell population under study, then check whether the signals occupy the same cells and whether IFITM1 follows the expected membrane pattern (HPA tissue IHC profile; HPA subcellular: plasma membrane supported). Choose fluorophores after checking unstained tissue for autofluorescence, and include single-stain controls to assess bleed-through (standard IF practice). Because the antibody epitope is unspecified, compare gentle permeabilisation with no permeabilisation before interpreting absent signal from the annotated cytoplasmic regions or the extracellular region (supplied antibody evidence: epitope unspecified; UniProt P13164 topology).
What should I check when IFITM1 chromogenic staining looks widespread?
Compare the stained section with a no-primary control and inspect whether colour follows cells, tissue folds or damaged edges (standard IHC practice). Apply a peroxidase block and assess endogenous enzyme signal before DAB development; these are general chromogenic IHC checks, not evidence specific to IFITM1 (standard IHC practice). If background remains high, titrate the catalog antibody and detection reagents on adjacent sections while keeping retrieval and development time consistent (standard IHC practice). Some signal across different cell populations is plausible because HPA describes low tissue specificity, but uniform deposits without cellular boundaries need independent scrutiny (HPA tissue RNA specificity; standard IHC practice).
How should I score IFITM1 staining across samples with different cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because IFITM1 can mark membranes in more than one tissue population (HPA tissue IHC profile; UniProt P13164 localisation). For each annotated region, report the percentage of positive cells and an intensity-based H-score using the same thresholds across slides (standard IHC practice). Normalise positive counts to the number of eligible cells, or report positive-cell density per mm² of viable annotated tissue when cell counting is impractical (standard IHC practice). Keep endothelial and lymphoid compartments separate where present, and exclude folds and necrosis from the denominator so their abundance does not drive an apparent group difference (HPA tissue IHC profile; standard IHC practice).
How can I distinguish true IFITM1 positivity from an IHC artefact?
A credible result combines staining in the appropriate cells with a membrane-associated pattern, while allowing plausible intracellular puncta at lysosome membranes (HPA tissue IHC profile; UniProt P13164 localisation). Check a known positive control such as epididymal glandular cells and compare the specimen with its no-primary control (HPA tissue IHC: high in epididymal glandular cells; standard IHC practice). Question colour restricted to cut edges, folds or necrotic areas, as these patterns require evaluation apart from intact-cell staining (standard IHC practice). Diffuse nuclear colour or signal retained in the no-primary control does not fit the annotated IFITM1 locations; investigate background and endogenous peroxidase before calling it positive (UniProt P13164 localisation; standard IHC practice).
Boster reagents
Best IFITM1 / Interferon-induced transmembrane protein 1 IHC Antibodies
Anti-IFITM1 antibodies have IHC data from human intestinal cancer tissue (PA1112 image caption) and IF/ICC data from Jurkat cells (A02633 image captions). Listed reactivity includes human, mouse and rat (catalog reactivity).
Real IHC data
Anti-IFITM1 Antibody ®
Cat # PA1112
Real IF data
Anti-IFITM1 Antibody
Cat # A02633
PA1112 will render with an IHC(P) image of human intestinal cancer tissue and lists human reactivity (PA1112 image caption; catalog reactivity). A02633 will render with IF/ICC images of Jurkat cells and lists human, mouse and rat reactivity (A02633 image captions; catalog applications and reactivity).
Which to pick: Choose PA1112 for paraffin-section tissue IHC, supported by its IHC(P) image of human intestinal cancer tissue; the fixative is unreported (PA1112 image caption). Choose A02633 for IF/ICC in Jurkat cells, supported by its images and application list (A02633 image captions; catalog applications). For mouse or rat samples, A02633 lists reactivity with those species, but the provided IF/ICC images show only Jurkat cells (A02633 catalog reactivity; image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P13164 (IFM1_HUMAN, Interferon-induced transmembrane protein 1).
- Human Protein Atlas. IFITM1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. IFITM1 subcellular location (ICC-IF): Localized to the plasma membrane and the Golgi apparatus..
- Human Protein Atlas. IFITM1 antibody validation summary (2 antibodies).
- Expression of IFITM1 as a prognostic biomarker in resected gastric and esophageal adenocarcinoma. Biomarker research 2016 — PMC4867989.
- Inflammatory pathways confer resistance to chemoradiotherapy in anal squamous cell carcinoma. NPJ precision oncology 2024 — PMC11039690.
- Interferon-Induced Transmembrane Protein 1 (IFITM1) Promotes Distant Metastasis of Small Cell Lung Cancer. International journal of molecular sciences 2020 — PMC7404048.
- Combination of IFITM1 knockdown and radiotherapy inhibits the growth of oral cancer. Cancer science 2018 — PMC6172064.
- PubMed PMID:2492664 — UniProt-cited evidence.
- PubMed PMID:7559564 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for IFITM1 (Interferon-induced transmembrane protein 1). Expected localisation Membranous staining in epididymal glands (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: PA1112 at 0.5-1μg/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 PA1112); 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-IFITM1, 0.5-1μg/ml (datasheet PA1112); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PA1112); Caveat: Endothelial and lymphoid positivity can complicate scoring (HPA tissue IHC); Isoform / epitope: No annotated isoforms; cytoplasmic vs extracellular epitope matters (UniProt); Regulation: Interferon induced expression (UniProt). Evidence: UniProt P13164, HPA tissue IHC · Enhanced, HPA ICC-IF · plasma membrane and the Golgi apparatus., Datasheet PA1112, PMC4867989, PMC7404048. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/IFITM1/immunohistochemistry-ihc-guide.