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TM4SF1 / Transmembrane 4 L6 family member 1 · IHC design guide
Design Immunohistochemistry for TM4SF1
Plan chromogenic TM4SF1 IHC in paraffin sections using lung alveolar cells as a staining reference (HPA tissue IHC). Start the catalog antibody at 5 μg/mL (datasheet: A10368), and assess membranous and cytoplasmic staining in light of the uncertain tissue evidence (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TM4SF1 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 and cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Alveolar and glandular cells show staining in a few tissues (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+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 paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Staining has low concordance with RNA expression (HPA tissue IHC) | |
| Regulation | Expression regulation is unreported (UniProt) | |
| Isoform / epitope | No annotated isoforms; epitope side still matters (UniProt) |
Section 1
Recommended TM4SF1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet) is accompanied by three published TM4SF1 IHC protocols (PMC6392956; PMC6010756; PMC12174346).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A10368); 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-TM4SF1, 5 μg/mL (datasheet A10368) |
| 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 | TM4SF1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic and membranous expression in a few tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval rule); use the article’s conditions when reproducing a published protocol.
Section 2
What Is the Expected TM4SF1 Staining Pattern?
TM4SF1 is a membrane protein with 4 transmembrane segments (UniProt P30408 topology). In paraffin-section IHC, expect membranous staining, potentially with cytoplasmic staining, in selected glandular or alveolar cells (HPA tissue IHC). HPA reports medium staining in several normal tissues, while UniProt also reports expression in some normal endothelial cells (HPA tissue IHC; UniProt P30408 tissue specificity). Interpret apparent positives cautiously: HPA rates its tissue IHC profile Uncertain because antibody staining and RNA data have low consistency (HPA tissue IHC reliability).
| Membranous staining in breast glandular cells or lung alveolar cells, with possible cytoplasmic staining (HPA tissue IHC). | This fits the reported compartment and cell distribution: both examples have medium staining in HPA, and UniProt places TM4SF1 at the membrane (HPA tissue IHC; UniProt P30408 subcellular location). Record the stained cell type and compartment separately; HPA's Uncertain reliability limits how confidently this pattern alone identifies TM4SF1 (HPA tissue IHC reliability). |
| Predominantly nuclear staining, without a convincing membrane-associated pattern (UniProt P30408 subcellular location). | Nuclear localisation is unsupported by the supplied TM4SF1 record; UniProt assigns the protein to membranes, and HPA describes cytoplasmic and membranous tissue staining (UniProt P30408; HPA tissue IHC). Treat an isolated nuclear signal as suspect and review counterstain alignment and detection controls (general IHC practice). |
| Staining in a cell population reported as not detected, such as adipocytes or marrow hematopoietic cells (HPA tissue IHC). | Check cell identification, cross-reactivity and endogenous detection activity before calling the signal TM4SF1 (general IHC practice). HPA lists those populations as not detected, but its Uncertain IHC reliability means a disagreement needs independent assessment rather than automatic rejection (HPA tissue IHC). |
| Diffuse staining across tissue and background, obscuring cell boundaries (general IHC practice). | A widespread haze cannot establish the compartment-specific pattern expected from UniProt membrane localisation and HPA tissue staining (UniProt P30408; HPA tissue IHC). Compare a no-primary control, wash adequacy and detection background, then judge staining only where cellular outlines remain interpretable (general IHC practice). |
| No detectable staining in breast glandular cells or lung alveolar cells (HPA tissue IHC). | HPA reports medium staining in these populations, so review section quality, antigen retrieval, antibody concentration and detection performance (HPA tissue IHC; general IHC practice). A negative result does not by itself establish biological absence: the reference tissue profile and both listed IHC antibody assessments are Uncertain (HPA tissue IHC; HPA antibodies). |
💡Expected TM4SF1 appearanceCall a convincing positive when selected glandular or alveolar cells show interpretable membranous staining, possibly with cytoplasmic staining, around HPA's reported medium level; isolated nuclear staining or uniform haze is suspect (HPA tissue IHC; UniProt P30408 subcellular location; general IHC practice).
| Compartment and topology (UniProt P30408 topology). | TM4SF1 has 4 membrane-spanning segments and extracellular as well as cytoplasmic regions (UniProt P30408 topology). Score membrane association first; HPA also reports cytoplasmic tissue staining, so cytoplasmic signal alone needs cautious interpretation (HPA tissue IHC). The supplied record does not locate either antibody's epitope (HPA antibodies). |
| Normal-tissue distribution (HPA tissue IHC). | HPA reports medium staining in adrenal, breast and fallopian-tube glandular cells, lung alveolar cells and pancreatic exocrine glandular cells (HPA tissue IHC). It reports selected other populations as low or not detected; compare like cell types rather than assigning one expected intensity to every tissue (HPA tissue IHC). |
| Antibody evidence (HPA antibodies). | HPA labels tissue IHC reliability Uncertain and lists IHC assessments as Uncertain for HPA002823 and CAB002760 (HPA tissue IHC; HPA antibodies). Reproducing a plausible pattern with one antibody does not resolve that uncertainty; use controls and an independent check when the interpretation is consequential (general IHC practice). |
| Glycosylation and processing (UniProt P30408). | UniProt annotates 2 glycosylation sites, at residues 129 and 159, and a single chain spanning residues 1–202; it annotates no signal peptide or propeptide (UniProt P30408). These features do not identify the antibodies' epitopes or establish a target-specific fixation or retrieval effect (UniProt P30408; HPA antibodies). |
| IF/ICC: what pattern is expected? (HPA subcellular ICC-IF). | For the separate IF/ICC guide, membrane localisation is the supported starting expectation (HPA subcellular ICC-IF; UniProt P30408). HPA supplies a membrane summary but no main-location assignment or cell-line ICC-IF images here; this record does not support an IF protocol choice or a validated cell-line example (HPA subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in an HPA medium-staining reference population (HPA tissue IHC). | The assay may have failed, or this section may differ from HPA's uncertain tissue observation (general IHC practice; HPA tissue IHC reliability). | Verify tissue identity and morphology, then review retrieval, antibody concentration and detection controls using the assay's documented conditions (general IHC practice). Do not infer a TM4SF1-specific retrieval failure from these sources (HPA tissue IHC; UniProt P30408). |
| Only nuclear staining appears (UniProt P30408 subcellular location). | The compartment conflicts with UniProt membrane localisation and HPA's cytoplasmic/membranous description (UniProt P30408; HPA tissue IHC). | Check counterstain alignment and no-primary background; reassess with an independent antibody or method if the finding matters (general IHC practice). Avoid scoring an isolated nuclear signal as the expected pattern (UniProt P30408; HPA tissue IHC). |
| A broad brown haze hides cell outlines (general IHC practice). | Non-specific staining or detection background may be obscuring cellular localisation (general IHC practice). | Compare the no-primary control and review blocking, washing and detection settings; score only interpretable cells (general IHC practice). HPA's reported cytoplasmic staining does not make tissue-wide haze a positive result (HPA tissue IHC). |
| Adipocytes or marrow hematopoietic cells appear strongly positive (HPA tissue IHC). | Cell misidentification, cross-reactivity or endogenous detection activity is possible; HPA reports these cells as not detected (general IHC practice; HPA tissue IHC). | Confirm the cell type and inspect appropriate detection controls before attributing staining to TM4SF1 (general IHC practice). Document persistent disagreement because HPA rates its IHC profile Uncertain (HPA tissue IHC reliability). |
| Cytoplasmic signal dominates, with little membrane definition (HPA tissue IHC). | HPA allows cytoplasmic staining, but a poorly resolved signal may also prevent a confident compartment call (HPA tissue IHC; general IHC practice). | Review morphology, staining concentration and detection background; report membrane and cytoplasmic components separately (general IHC practice). Do not claim cytoplasmic staining alone proves specificity under HPA's Uncertain assessment (HPA tissue IHC reliability). |
| Endothelial staining appears beside epithelial staining (UniProt P30408 tissue specificity). | Some normal endothelial expression is reported by UniProt, so this distribution can have a biological explanation (UniProt P30408 tissue specificity). | Identify endothelial cells morphologically and score them separately from glandular or alveolar cells (general IHC practice; HPA tissue IHC). Check controls before treating every vascular signal as specific, given HPA's Uncertain tissue IHC reliability (HPA tissue IHC). |
Experiment evidence
IHC & IF images showing TM4SF1 staining
A10368 · IHCImmunohistochemistry of TM4SF1 in human lung tissue with TM4SF1 antibody at 5 μg/mL.
A10368 · IHCImmunohistochemistry of TM4SF1 in mouse lung tissue with TM4SF1 antibody at 5 μg/mL.
A10368 · IFImmunofluorescence of TM4SF1 in human lung tissue with TM4SF1 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000169908-TM4SF1/tissue/cerebral+cortex#imid_1008552
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000169908-TM4SF1/tissue/colon#imid_1008494
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000169908-TM4SF1/tissue/liver#imid_1008487
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000169908-TM4SF1/tissue/kidney#imid_1008488
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TM4SF1 IHC & IF
🧪Run lung first: alveolar cells should stain (HPA: Medium in lung alveolar cells). Use bone marrow hematopoietic cells as the negative tissue (HPA: Not detected); on the lung slide, assess background in neighboring unstained cells without assuming endothelial cells are negative (UniProt P30408: present on some normal endothelial cells).
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for TM4SF1; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and host- and clonality-matched isotype controls, plus TM4SF1 knockout material where available or immunizing-peptide competition if the peptide is available (standard IHC practice). For chromogenic lung IHC, quench endogenous peroxidase and check for endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; optimize antigen retrieval empirically (standard IHC practice). The selected lung IHC caption reports 5 µg/mL antibody but leaves the fixative unreported (A10368 caption). Whether frozen sections or IF/ICC are easier is unreported; lung autofluorescence can complicate IF interpretation (standard IF practice).
HPA tissue IHC evidence for TM4SF1
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Medium | Protein (IHC) | HPA → |
| Lung | Alveolar cells | Medium | Protein (IHC) | HPA → |
| Pancreas | Exocrine 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 → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic 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 TM4SF1 IHC Tips
Troubleshoot TM4SF1 chromogenic IHC by checking retrieval, tissue context, membrane localisation and staining controls before scoring (UniProt P30408; HPA tissue IHC).
What retrieval conditions should I start with for weak TM4SF1 staining?
Use citrate pH 6.0 heat-induced antigen retrieval at 95–98 °C for 20 min (page retrieval rule). TM4SF1 spans the membrane four times, with extracellular and cytoplasmic regions whose accessibility may differ (UniProt P30408 topology). Run a known staining control alongside each retrieval condition; the selected antibody has a human lung tissue IHC image at 5 µg/mL (caption: A10368). If staining remains weak, compare a carefully controlled EDTA pH 9 retrieval trial with citrate, keeping detection conditions constant (standard IHC practice). Judge improvement by cellular staining and preserved tissue morphology, since stronger diffuse colour alone does not establish specificity (standard IHC practice).
How should I investigate a weak result after fixation?
Target-specific sensitivity of TM4SF1 to fixation is unknown from the supplied evidence; the selected tissue-IHC caption does not state its fixative (caption: A10368). Record the fixative, fixation duration and processing history for each section before comparing staining across blocks (standard IHC practice). Compare sections processed in the same batch using the page's citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval rule). Include a known staining control and a primary-antibody omission control to separate weak antigen detection from detection-system failure (standard IHC practice). Interpret any fixation effect as an experimental observation requiring matched controls, rather than inferring it from topology or tissue staining patterns (standard IHC practice).
Which staining compartment is plausible for TM4SF1 in tissue sections?
Expect a membrane-associated pattern because TM4SF1 is annotated as a membrane protein with four transmembrane segments (UniProt P30408 topology). The tissue-IHC profile also reports cytoplasmic and membranous expression in a few tissues, although its reliability is uncertain (HPA tissue IHC). Examine whether colour outlines identifiable cells or is confined to plausible intracellular regions, and compare those patterns across adjacent sections (standard IHC practice). Endothelial cells are a plausible positive population, while HPA reports medium staining in lung alveolar cells (UniProt P30408 tissue specificity; HPA: Lung, alveolar cells, Medium). Diffuse extracellular precipitate or staining detached from intact cells should prompt a control review before being called TM4SF1 positive (standard IHC practice).
How can epitope position change the staining pattern?
Check the antibody's documented immunogen or epitope before interpreting a membrane-negative result; its position is not specified in the supplied caption (caption: A10368). TM4SF1 has extracellular regions at residues 31–49 and 115–161, plus cytoplasmic regions at 1–9, 71–93 and 183–202 (UniProt P30408 topology). Glycosylation sites at residues 129 and 159 could matter when assessing accessibility of an extracellular epitope, but no effect on this antibody is established (UniProt P30408 glycosylation). The record annotates no isoforms, so it provides no basis for assigning different tissue patterns to isoform-specific staining (UniProt P30408 isoforms). Compare localisation under matched retrieval and detection conditions before attributing a pattern change to epitope accessibility (standard IHC practice).
How should I follow up an IHC pattern using multiplex IF?
Use the IHC result to select regions for IF, then pair TM4SF1 with an endothelial marker such as CD31 when assessing endothelial staining (UniProt P30408 tissue specificity; standard IF practice). Choose spectrally separated fluorophores and place the weaker signal in a far-red channel after checking unstained tissue for autofluorescence (standard IF practice). If the antibody recognises a cytoplasmic region, compare permeabilised and non-permeabilised preparations; an extracellular epitope may be accessible without permeabilisation (UniProt P30408 topology; standard IF practice). The supplied evidence identifies no ICC/IF images for this target, so establish IF specificity with appropriate single-stain and primary-omission controls (HPA subcellular; standard IF practice). Compare cell-level IF localisation with adjacent chromogenic sections without assuming that the IHC tissue caption validates IF conditions (caption: A10368; standard IF practice).
What should I check when TM4SF1 staining is diffuse or widespread?
First compare the stained section with a primary-antibody omission control to identify signal arising from the detection system (standard IHC practice). In a peroxidase/DAB workflow, check endogenous peroxidase blocking and inspect whether colour appears in damaged tissue or outside cell boundaries (standard IHC practice). Titrate the primary antibody around the reported 5 µg/mL tissue-IHC image condition while holding retrieval and detection constant (caption: A10368; standard IHC practice). Review blocking, wash stringency and section drying when background covers both expected and unexpected tissue compartments (standard IHC practice). HPA rates its tissue-IHC evidence uncertain, so widespread colour should be assessed against controls and morphology before being assigned to TM4SF1 (HPA tissue IHC).
How should I score heterogeneous TM4SF1 staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cellular compartment and eligible cell population before scoring, using membrane-associated staining as the primary localisation reference (UniProt P30408 subcellular location; standard IHC practice). Record the percentage of positive viable cells and an intensity category, then calculate an H-score from 0–300 if intensity grading is reproducible (standard IHC practice). For spatial comparisons, report positive cells per mm² of viable tissue and retain the analysed area as the denominator (standard IHC practice). Separate endothelial from other cellular staining where morphology permits, because endothelial expression is reported for TM4SF1 (UniProt P30408 tissue specificity). Keep retrieval, imaging and thresholds consistent across sections, and document excluded necrotic or damaged regions (standard IHC practice).
When should an apparent TM4SF1-positive region be treated cautiously?
Give greatest weight to staining associated with intact cells and a plausible membrane pattern, consistent with TM4SF1's annotated location (UniProt P30408 subcellular location; standard IHC practice). Check the stained cell type: endothelial cells are plausible, and HPA reports medium staining in lung alveolar cells (UniProt P30408 tissue specificity; HPA: Lung, alveolar cells, Medium). Question colour concentrated at section edges, in necrotic areas or outside cell boundaries, where processing artefacts can occur (standard IHC practice). If a peroxidase/DAB workflow is used, compare primary-omission controls to assess endogenous enzyme or detection-related colour (standard IHC practice). Treat an unexpected compartment or cell population as provisional until it agrees with controls, morphology and independent evidence, given HPA's uncertain tissue-IHC reliability (HPA tissue IHC; standard IHC practice).
Boster reagents
Best TM4SF1 / Transmembrane 4 L6 family member 1 IHC Antibodies
A10368 has IHC images from human and mouse lung and IF images from human and mouse lung (catalog image captions); rat reactivity is listed without an image (catalog reactivity).
Real IHC data
Anti-TM4SF1 Antibody
Cat # A10368
A10368 will render with an IHC image from human lung at 5 μg/mL (catalog IHC image caption). Its additional images show IHC in mouse lung and IF in human and mouse lung (catalog image captions).
Which to pick: Choose A10368 for paraffin section IHC because IHC-P is listed and its own IHC images show staining in human and mouse lung (catalog applications; catalog IHC image captions); the fixative is unreported (catalog IHC image captions). For tissue IF, A10368 has human and mouse lung images at 20 μg/mL (catalog IF image captions); ICC validation is unreported (catalog applications). For work across species, A10368 lists human, mouse and rat reactivity, with IHC images for human and mouse only (catalog reactivity; catalog IHC image captions); clonality is unreported (catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P30408 (T4S1_HUMAN, Transmembrane 4 L6 family member 1).
- Human Protein Atlas. TM4SF1 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. TM4SF1 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. TM4SF1 antibody validation summary (2 antibodies).
- Upregulation of transmembrane 4 L6 family member 1 predicts poor prognosis in invasive breast cancer: A STROBE-compliant article. Medicine 2017 — PMC6392956.
- Clinical significance of TM4SF1 as a tumor suppressor gene in gastric cancer. Cancer medicine 2018 — PMC6010756.
- TM4SF1 promotes EMT and cancer stemness via the Wnt/β-catenin/SOX2 pathway in colorectal cancer. Journal of experimental & clinical cancer research : CR 2020 — PMC7643364.
- Bladder cancer variants share aggressive features including a CA125+ cell state and targetable TM4SF1 expression. Nature communications 2025 — PMC12174346.
- PubMed PMID:1565644 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:7510285 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TM4SF1 (Transmembrane 4 L6 family member 1). Expected localisation Membranous and cytoplasmic staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A10368 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (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 A10368); 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-TM4SF1, 5 μg/mL (datasheet A10368); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Staining has low concordance with RNA expression (HPA tissue IHC); Isoform / epitope: No annotated isoforms; epitope side still matters (UniProt); Regulation: Expression regulation is unreported (UniProt). Evidence: UniProt P30408, HPA tissue IHC · Uncertain, HPA ICC-IF · Membrane, Datasheet A10368, PMC6392956, PMC6010756. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TM4SF1/immunohistochemistry-ihc-guide.