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IFITM1 / Interferon-induced transmembrane protein 1 · IHC design guide
Design Immunohistochemistry for IFITM1
Plan chromogenic IHC on paraffin sections using epididymal glandular membrane staining as a positive reference (HPA tissue IHC). The catalog antibody’s IHC-P dilution is 0.5–1 μg/mL (datasheet: PA1112).
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 | Epididymal glandular cell membrane (HPA tissue IHC) | |
| Staining pattern | Distinct membranous staining in epididymal glandular cells (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 staining can confound scoring (HPA tissue IHC) | |
| Regulation | Interferon inducible (UniProt) | |
| Isoform / epitope | No annotated isoforms; epitope side matters (UniProt) |
Section 1
Recommended IFITM1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with four published IFITM1 IHC protocols (datasheet; PMC4867989; PMC11039690; 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 antigen-retrieval rule); published alternatives use high-pH heat or trypsin (PMC11039690; PMC6172064).
Section 2
What Is the Expected IFITM1 Staining Pattern?
IFITM1 is a membrane protein with a transmembrane segment at residues 87–107 (UniProt P13164 topology). In paraffin sections, expect prominent membranous staining in epididymal glandular cells (HPA: High; IHC reliability Enhanced). Endothelial cells and lymphoid tissue may also stain (HPA: tissue IHC profile).
| Strong glandular cell membrane staining in epididymis (HPA: High). | Expected positive pattern (HPA: distinct membranous expression). |
| Medium staining in lymph node germinal center cells (HPA: Medium). | Compatible with the observed tissue pattern (HPA: lymph node IHC). |
| Predominantly nuclear staining (UniProt P13164: membrane localization). | Unexpected compartment; assess artifact or antibody specificity (UniProt P13164). |
| Staining in adipocytes (HPA: Not detected). | Unexpected cell type; assess cross-reactivity or endogenous detection activity (HPA: adipose IHC). |
| No glandular cell signal in epididymis (HPA: High). | Unexpected in this positive control; check assay performance (HPA: epididymis IHC). |
💡Expected IFITM1 appearanceA positive section shows High membranous signal in epididymal glandular cells; nuclear or diffuse staining is suspect (HPA: epididymis IHC; UniProt P13164).
| Localization (UniProt P13164; HPA: subcellular). | Cell membrane is expected; lysosome membrane is annotated; Golgi is uncertain (UniProt P13164; HPA: ICC-IF). |
| Epitope position (UniProt P13164 topology). | The extracellular region is residues 108–125; antibody epitope is unspecified (UniProt P13164). |
| Tissue control (HPA: tissue IHC). | Epididymal glandular cells are High; adipocytes are Not detected (HPA: tissue IHC). |
| IHC antibody validation (HPA: antibodies). | HPA004810 and CAB017615 are both IHC Enhanced (HPA: antibody validation). |
| IF/ICC expected pattern? (HPA: subcellular). | Plasma membrane is supported; Golgi localization is uncertain (HPA: ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in epididymal glandular cells (HPA: High). | Detection or staining failure is possible (general IHC practice). | Check positive control and detection reagents (general IHC practice). |
| Predominantly nuclear signal (UniProt P13164: membrane localization). | Nonspecific staining is possible (general IHC practice). | Review antibody controls and cell boundaries (general IHC practice). |
| Staining in adipocytes (HPA: Not detected). | Cross-reactivity or endogenous activity is possible (general IHC practice). | Compare antibody-omission and tissue controls (general IHC practice). |
| Diffuse staining across the section (HPA: distinct membranous pattern). | Background from detection or inadequate blocking is possible (general IHC practice). | Check controls, blocking and wash steps (general IHC practice). |
| Weak signal in epididymis (HPA: glandular cells High). | Assay sensitivity may be insufficient (general IHC practice). | Review retrieval, antibody dilution and detection controls (general IHC practice). |
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 a negative comparator (HPA: adipocytes Not detected); on the epididymis slide, use adjacent nonglandular cells as a background comparator only if they are unstained.
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 no-primary (secondary-only) and isotype controls matched to the primary antibody’s host species and class, with a comparable monoclonal or polyclonal preparation where possible (standard IHC practice). Use IFITM1 knockout material as a biological negative if available (standard IHC practice). For chromogenic epididymis sections, block endogenous peroxidase and check endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and any fixation effect are unreported in the supplied evidence; the selected PA1112 IHC(P) caption also leaves the fixative unreported (PA1112 caption: fixative not stated). Retrieval dependency and whether frozen sections or IF are easier are unreported, so optimize retrieval empirically for paraffin IHC (standard IHC practice). In epididymis, distinguish glandular-cell staining from luminal deposits using the no-primary control (standard IHC practice).
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
Troubleshoot IFITM1 staining in paraffin sections by checking retrieval, compartment, cell type, controls and scoring before interpreting chromogenic signal.
How should I retrieve IFITM1 in paraffin sections when staining is weak?
Start with citrate buffer at pH 6.0, using heat induced epitope retrieval at 95–98 °C for 20 minutes (page retrieval setting). Let sections cool consistently, then compare the same antibody dilution and detection conditions across a retrieval time series around 20 minutes (general IHC practice). IFITM1 has a membrane spanning segment at residues 87–107, but that topology alone does not predict which retrieval condition exposes this antibody’s epitope (UniProt P13164 topology). If staining remains weak, test another buffer or pH as a fallback on matched sections, judging signal against tissue morphology and negative controls (general IHC practice).
Could fixation explain weak or uneven IFITM1 staining?
Target specific fixation sensitivity is unknown because no fixation condition is reported for the selected PA1112 tissue IHC image (PA1112 tissue IHC caption). Record the fixative, fixation interval, processing schedule and section age for each sample before comparing staining intensity (general IHC practice). On matched sections, hold retrieval at citrate pH 6.0 and 95–98 °C for 20 minutes while checking whether differences track processing batches (page retrieval setting; general IHC practice). The caption identifies human intestinal cancer tissue and IHC(P), but supplies no basis for attributing staining differences to a particular fixative (PA1112 tissue IHC caption).
Which IFITM1 staining pattern should I expect in tissue sections?
Prioritise membranous staining when assessing IFITM1 in chromogenic sections: the protein is assigned to cell and lysosome membranes, and tissue IHC describes distinct membranous expression in epididymis (UniProt P13164 subcellular location; HPA tissue IHC profile). Intracellular puncta may warrant review because lysosome membrane localisation is annotated, while a supported plasma membrane and uncertain Golgi location are reported in cell imaging (UniProt P13164 subcellular location; HPA subcellular). Compare apparent membrane signal with adjacent cells at the same magnification and with the counterstain clearly visible (general IHC practice). Diffuse nuclear staining should prompt a control and detection review before it is scored as IFITM1 (UniProt P13164 subcellular location; general IHC practice).
How can epitope position affect IFITM1 IHC results?
Ask which residues the antibody recognises before interpreting a weak or altered pattern; the supplied evidence does not identify the catalog antibody’s epitope (supplied antibody evidence). IFITM1 is annotated as a 125 residue chain with a membrane spanning segment at 87–107, cytoplasmic regions at 1–36 and 58–86, and an extracellular region at 108–125 (UniProt P13164 topology). A phosphoserine is annotated at residue 16, while no glycosylation sites or isoforms are annotated; these facts do not establish antibody recognition or rule out every biological variant (UniProt P13164 record). Compare antibody staining with an independent assay or antibody targeting a documented, distinct epitope when specificity remains uncertain (general IHC practice).
How should I check IFITM1 localisation by multiplex immunofluorescence?
Use IF/ICC as a separate validation experiment and pair IFITM1 with a marker for the expected cell population, such as glandular cells in epididymis or endothelial cells where tissue IHC reports expression (HPA tissue IHC profile). Choose spectrally separated fluorophores and inspect unstained tissue so autofluorescence does not masquerade as membrane signal (general IF practice). Match permeabilisation to the documented antibody epitope: a cytoplasmic epitope may require membrane access, whereas excessive permeabilisation can complicate assessment of a surface facing epitope (UniProt P13164 topology; general IF practice). Include single stain and secondary only controls, then compare membrane and intracellular patterns with the reported plasma membrane and lysosome membrane locations (general IF practice; HPA subcellular; UniProt P13164 subcellular location).
How can I reduce background without losing IFITM1 membrane signal?
First distinguish diffuse background from staining in expected compartments and cell populations, including epididymal glandular cells and endothelial cells (HPA tissue IHC profile; UniProt P13164 subcellular location). Run a no primary control and check endogenous peroxidase blocking before interpreting DAB deposits as IFITM1 (general chromogenic IHC practice). If the control is clean but tissue staining is widespread, titrate the primary antibody and shorten chromogen development on matched sections while keeping citrate pH 6.0 retrieval constant (general IHC practice; page retrieval setting). Assess section edges, folds and damaged areas separately, and retain counterstain detail so weak membrane signal is distinguishable from nonspecific deposits (general IHC practice).
What is a defensible way to quantify IFITM1 IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because HPA tissue IHC reports membranous epididymal expression and staining in endothelial and lymphoid tissue (HPA tissue IHC profile). For a defined population, report the percentage of positive cells and an H-score based on the proportion at each intensity; use positive cell density per mm² when cell abundance is the question (general IHC quantification practice). Normalise cell counts to the analysed tissue area or the total number of eligible cells, and keep thresholds and image settings fixed across samples (general IHC quantification practice). Exclude folds, necrosis and section edges by preset rules, and report membrane scoring separately from any intracellular staining (general IHC quantification practice; UniProt P13164 subcellular location).
How can I distinguish true IFITM1 positivity from staining artefact?
Look for reproducible staining in a plausible compartment and cell type: membrane signal in epididymal glandular cells is strongly supported, while endothelial and lymphoid expression is also reported (HPA tissue IHC profile; UniProt P13164 subcellular location). Compare suspect staining with a no primary control and with a tissue region expected to have little or no signal, such as adipocytes in adipose tissue (general IHC practice; HPA tissue IHC negative profile). Nuclear dominant signal, edge restricted staining and deposits over necrosis deserve investigation before a positive call (UniProt P13164 subcellular location; general IHC practice). Check endogenous peroxidase blocking and DAB development when brown deposits persist without primary antibody, then interpret remaining signal alongside morphology and matched controls (general chromogenic IHC practice).
Boster reagents
Best IFITM1 / Interferon-induced transmembrane protein 1 IHC Antibodies
Anti-IFITM1 antibodies have real IHC data from human intestinal cancer tissue (PA1112 image caption) and IF/ICC data from human Jurkat cells (A02633 image captions).
Real IHC data
Anti-IFITM1 Antibody ®
Cat # PA1112
Real IF data
Anti-IFITM1 Antibody
Cat # A02633
PA1112 is listed for human IHC and shows paraffin-section staining of human intestinal cancer tissue (PA1112 applications, reactivity and image caption). A02633 shows IF/ICC in human Jurkat cells; its listed reactivity also includes Mouse and Rat (A02633 image captions and catalog reactivity).
Which to pick: Choose PA1112 for human paraffin-section IHC based on its IHC(P) image (PA1112 image caption); the fixative is unreported (PA1112 image caption). Choose A02633 for IF/ICC based on its listed applications and Jurkat cell images, or when listed Mouse or Rat reactivity is needed (A02633 applications, image captions and catalog reactivity). Neither entry reports a clone, so clonality does not distinguish them (catalog clone fields).
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 Epididymal glandular cell membrane (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 staining can confound scoring (HPA tissue IHC); Isoform / epitope: No annotated isoforms; epitope side matters (UniProt); Regulation: Interferon inducible (UniProt). Evidence: UniProt P13164, HPA tissue IHC · Enhanced, HPA ICC-IF · plasma membrane and the Golgi apparatus., Datasheet PA1112, PMC4867989, PMC11039690. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/IFITM1/immunohistochemistry-ihc-guide.