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SLC14A1 / Urea transporter 1 · IHC design guide
Design Immunohistochemistry for SLC14A1
Plan paraffin IHC for SLC14A1 with the catalog antibody at 2–5 μg/ml (datasheet A04926-2). Use erythrocyte staining as a reference (HPA tissue IHC) and assess its cytoplasmic pattern alongside the expected membrane location (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SLC14A1 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 stain (HPA tissue IHC); membrane expected (UniProt) | |
| Staining pattern | Cytoplasmic staining, most abundant in erythrocytes (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04926-2) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining has low concordance with RNA (HPA tissue IHC) | |
| Regulation | Regulation not established (UniProt) | |
| Isoform / epitope | Two isoforms; map extracellular vs cytoplasmic epitopes (UniProt) |
Section 1
Recommended SLC14A1 IHC & IF Protocols
The catalog antibody's IHC-P protocol (datasheet A04926-2) is accompanied by published SLC14A1 IHC methods for urothelial, colorectal and prostate specimens (PMC7546005; PMC11282742; PMC9579171).
| Sample | Paraffin-embedded human melanoma tissue; fixative not specified (datasheet A04926-2) |
| Fixation | Image fixative and duration unreported (datasheet A04926-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A04926-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04926-2) |
| Primary antibody | Rabbit anti-SLC14A1, 2-5 μg/ml (datasheet A04926-2) |
| Primary incubation | Overnight at 4 °C (datasheet A04926-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04926-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SLC14A1-positive staining in processes in granular layer of cerebellum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues, most abundant in erythrocytes. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet A04926-2); the prostate study used 10 mM citrate at pH 6.0 (PMC9579171).
Section 2
What Is the Expected SLC14A1 Staining Pattern?
SLC14A1 is a nine-pass membrane transporter; UniProt places it at the cell membrane and restricts it to the basolateral membrane in portions of urothelium (UniProt Q13336 topology and subcellular location). On IHC, expect conspicuous erythrocyte staining and assess cell-specific patterns such as colon enterocyte microvilli and placental endothelium (HPA tissue IHC). HPA rates the tissue IHC antibody Approved but reports low consistency between staining and RNA expression (HPA tissue IHC).
| Distinct staining outlines erythrocytes or follows the microvilli of colon enterocytes. | This fits the erythrocyte-rich tissue profile and High colon microvillar signal (HPA tissue IHC). Interpret a discrete surface pattern alongside tissue anatomy; UniProt places SLC14A1 at membranes but does not establish which IHC signal in a mixed section is specific (UniProt Q13336 subcellular location; HPA tissue IHC: low RNA–staining consistency). |
| Staining is predominantly nuclear in the cells being scored. | Nuclear dominance conflicts with the cell-membrane assignment and supported plasma-membrane ICC-IF localization (UniProt Q13336 subcellular location; HPA subcellular ICC-IF). Treat it as an unexpected pattern and review controls before calling the cells positive; HPA also describes cytoplasmic tissue staining, so a cytoplasmic component alone is not proof of artefact (HPA tissue IHC). |
| Strong staining appears in adipocytes or bone-marrow hematopoietic cells. | Those cell populations were Not detected in the supplied HPA tissue IHC observations (HPA tissue IHC). Consider cross-reactivity or endogenous chromogen activity, then compare a reagent-omission control and a known-positive section (general IHC practice). An HPA negative is an observed result under its conditions, not proof that every specimen must be negative (HPA tissue IHC). |
| Color spreads across cells and extracellular areas without a defined cellular pattern. | Diffuse background prevents reliable compartment or cell-type assignment. Excess primary or detection reagent, incomplete blocking, and inadequate washes are general IHC possibilities (general IHC practice). Do not score background as SLC14A1 merely because the tissue contains a listed positive cell population (HPA tissue IHC). |
| No signal appears in colon enterocyte microvilli or placental endothelium. | Both are listed as High in HPA tissue IHC (HPA tissue IHC). If an appropriate positive section fails in the same run, first assess section integrity, retrieval, antibody dilution, and detection controls (general IHC practice). A failed positive control makes a negative study section uninterpretable; it does not establish absent SLC14A1 expression (general IHC practice). |
💡Expected SLC14A1 appearanceCall a positive result when staining is cell-associated and anatomically resolved—for example, conspicuous erythrocytes or High colon enterocyte microvilli—while treating dominant nuclear color or diffuse cell-free color as suspect (HPA tissue IHC; UniProt Q13336 subcellular location; general IHC practice).
| Membrane topology and compartment | SLC14A1 has 9 annotated transmembrane segments and a cell-membrane assignment (UniProt Q13336 topology and subcellular location). Use cell outlines and polarized epithelial surfaces as localization clues; the record does not identify the antibody epitope, so topology alone cannot prescribe permeabilisation or retrieval (UniProt Q13336 topology; general IHC practice). |
| Cell-specific tissue evidence | HPA lists High colon enterocyte microvilli, placental endothelial cells, and cerebellar granular-layer processes; its overall profile says staining is most abundant in erythrocytes (HPA tissue IHC). Choose and score a defined structure rather than treating every cell in one tissue as an expected positive (HPA tissue IHC; general IHC practice). |
| Agreement between staining and RNA | The HPA tissue IHC reliability label is Approved, with low consistency between antibody staining and RNA expression and staining in unannotated cells or structures (HPA tissue IHC). That caveat calls for cautious interpretation of an unexpected cell population, even when the slide has a visually strong signal (HPA tissue IHC). |
| Glycosylation and isoforms | UniProt lists one glycosylation site at residue 211 and 2 isoforms (UniProt Q13336). These annotations do not show whether either feature changes this antibody’s staining, epitope access, or performance in paraffin sections; antibody-specific evidence would be needed for that interpretation (UniProt Q13336; general IHC practice). |
| IF/ICC Q: What localization can be expected? | A: HPA reports supported plasma-membrane localization in ICC-IF images from RT-4 using an ICC-Supported antibody; its IHC-Approved antibody is a different listed reagent (HPA subcellular ICC-IF; HPA antibodies). Use that result as compartment context for IHC, without assuming the two antibody validations are interchangeable (HPA antibodies). |
| Situation | Likely cause | Next action |
|---|---|---|
| The known-positive section has no visible signal. | A run-level failure is possible when a structure listed as High, such as colon enterocyte microvilli, is also blank (HPA tissue IHC; general IHC practice). | Check tissue preservation and the detection run, then review the catalog antibody’s IHC-P instructions for the specified retrieval and dilution; change one general IHC variable at a time (general IHC practice). |
| Study tissue is negative, but the positive control stains as expected. | Expression may be absent or below the assay’s detection limit in the scored cells; HPA staining varies by cell type and tissue (HPA tissue IHC; general IHC practice). | Report the scored cell population and control result. Avoid inferring a tissue-wide negative from one compartment, and compare the relevant HPA cell-level observation when available (HPA tissue IHC; general IHC practice). |
| Adipocytes or marrow hematopoietic cells stain strongly. | HPA lists these populations as Not detected; nonspecific binding or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Run a primary-antibody omission control, inspect whether color persists, and review blocking and detection reagents before assigning target positivity (general IHC practice). |
| Nuclear color dominates while cell surfaces are indistinct. | The pattern conflicts with UniProt’s membrane location and HPA’s supported plasma-membrane ICC-IF localization (UniProt Q13336 subcellular location; HPA subcellular ICC-IF). | Check a positive tissue structure and reagent-omission control, then review antibody dilution and detection background (HPA tissue IHC; general IHC practice). Do not use nuclear intensity alone as the positive score (UniProt Q13336 subcellular location). |
| Diffuse chromogen obscures enterocyte microvilli or endothelial borders. | Excess reagent, incomplete blocking, or insufficient washing can obscure cellular detail (general IHC practice). | Compare the reagent-omission control, review blocking and washes, and adjust detection or antibody concentration within the validated IHC workflow (general IHC practice). Score only a pattern that resolves the relevant cells or structures (HPA tissue IHC). |
| A strong tissue pattern conflicts with the expected cell population. | HPA explicitly reports low consistency between antibody staining and RNA expression and notes staining in unannotated cells or structures (HPA tissue IHC). | Document the exact cells and compartment, check controls, and avoid treating intensity alone as validation (HPA tissue IHC; general IHC practice). If testing another antibody, review its application-specific validation independently (HPA antibodies). |
Experiment evidence
IHC & IF images showing SLC14A1 staining
A04926-2 · IHCIHC analysis of UT-B/SLC14A1 using anti-UT-B/SLC14A1 antibody (A04926-2). UT-B/SLC14A1 was detected in a paraffin-embedded section of human
A04926-2 · IHCIHC analysis of UT-B/SLC14A1 using anti-UT-B/SLC14A1 antibody (A04926-2). UT-B/SLC14A1 was detected in a paraffin-embedded section of human
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000141469-SLC14A1/tissue/cerebral+cortex#imid_16462643
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000141469-SLC14A1/tissue/colon#imid_16459473
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000141469-SLC14A1/tissue/liver#imid_16459466
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000141469-SLC14A1/tissue/kidney#imid_16459475
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA077233; green = SLC14A1, blue = nucleus, red = microtubules.
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA077233; green = SLC14A1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SLC14A1 IHC & IF
🧪Run colon first: enterocyte microvilli should stain strongly (HPA: High in colon enterocytes, microvilli). Use adipose tissue as the negative, with adipocytes at background (HPA: Not detected in adipocytes); on the colon slide, compare staining with adjacent non-enterocyte cells as internal background controls (HPA: High in enterocyte microvilli).
Positive control tissue: Cerebellum (Processes in granular layer, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SLC14A1 in RT-4, with annotated localisation: Plasma membrane (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (selected-SKU caption: rabbit primary antibody). Use validated SLC14A1-knockout material or antigen-peptide competition as a biological specificity control; block endogenous peroxidase before HRP–DAB detection (selected-SKU caption: HRP–DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption: fixative unreported). That caption uses heat-mediated retrieval in EDTA at pH 8.0, but does not establish that retrieval is required (selected-SKU caption: EDTA retrieval). The supplied evidence does not establish whether frozen sections or IF are easier; colon microvilli require careful assessment of surface-localized staining against background (HPA: High in enterocyte microvilli).
HPA tissue IHC evidence for SLC14A1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. Antibody staining in cells/structures not annotated, view images.). 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 |
|---|---|---|---|---|
| Cerebellum | Processes in granular layer | High | Protein (IHC) | HPA → |
| Colon | Enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Placenta | Endothelial cells | High | Protein (IHC) | HPA → |
| Appendix | Enterocytes | Medium | Protein (IHC) | HPA → |
| Duodenum | Goblet cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SLC14A1 IHC Tips
Troubleshoot SLC14A1 chromogenic IHC in paraffin sections using the catalog antibody’s tissue image, protein annotations, and tissue staining data.
What retrieval should I try first when SLC14A1 staining is weak?
Start with heat-mediated antigen retrieval in EDTA, pH 8.0, for paraffin sections (datasheet A04926-2). The catalog antibody’s tissue image used this retrieval before incubation with 2 μg/ml primary antibody overnight at 4°C (datasheet A04926-2). SLC14A1 has 9 transmembrane segments, so assess membrane detail alongside overall signal when adjusting retrieval conditions (UniProt Q13336 topology; standard IHC practice). Keep section thickness, detection, and exposure to retrieval heat consistent across comparison slides (standard IHC practice). If staining remains weak, vary retrieval duration on matched sections while checking that tissue morphology and negative-control staining remain acceptable (standard IHC practice).
Could fixation explain variable SLC14A1 staining between paraffin blocks?
Target-specific fixation sensitivity is unknown from the supplied evidence: the catalog image identifies a paraffin-embedded section but does not state its fixative (datasheet A04926-2). Record each block’s fixative and processing history before attributing a signal difference to SLC14A1 abundance (standard IHC practice). Compare blocks with the same EDTA, pH 8.0, retrieval and the same primary-antibody incubation to limit procedural variation (datasheet A04926-2; standard IHC practice). Examine morphology and a positive-control section processed alongside each batch, because poor preservation can complicate cellular scoring (standard IHC practice). Do not use staining differences alone to infer a target-specific fixation effect (standard IHC practice).
Should SLC14A1 staining appear on membranes or in cytoplasm?
Prioritize a membrane-associated pattern: SLC14A1 is annotated at the cell membrane and restricted to the basolateral membrane in various portions of urothelium (UniProt Q13336 subcellular location). The HPA subcellular record also supports plasma-membrane localisation (HPA subcellular). Interpret that expectation with care because HPA tissue IHC reports cytoplasmic staining in several tissues, most abundant in erythrocytes (HPA tissue IHC). In paraffin sections, inspect cellular boundaries at high magnification and identify erythrocytes before assigning signal to neighboring cells (UniProt Q13336 tissue specificity; standard IHC practice). Compare the compartment and cell type against a matched negative control rather than treating diffuse chromogen as sufficient evidence (standard IHC practice).
How do I assess whether the antibody detects both SLC14A1 isoforms?
SLC14A1 has 2 annotated isoforms, but the supplied catalog image does not identify the antibody’s epitope or establish isoform coverage (UniProt Q13336 isoforms; datasheet A04926-2). Its 9 transmembrane segments and glycosylation site at residue 211 make epitope position relevant when interpreting staining, without showing that either feature changes this antibody’s signal (UniProt Q13336 topology and glycosylation). Obtain the antibody’s mapped immunogen or epitope before claiming isoform-specific detection (standard IHC practice). Compare sequence coverage across both isoforms, then assess staining with appropriate positive and negative tissue controls (UniProt Q13336 isoforms; standard IHC practice). Report the result as SLC14A1 immunoreactivity unless isoform discrimination is independently established (standard IHC practice).
How can I check an SLC14A1 IHC pattern by multiplex immunofluorescence?
For a separate IF experiment, pair SLC14A1 with a marker that identifies the expected cell population, such as an erythrocyte marker when evaluating erythrocyte-rich regions (UniProt Q13336 tissue specificity; standard IF practice). Choose fluorophores after measuring tissue autofluorescence and place the weaker signal in a cleaner spectral channel (standard IF practice). Select permeabilisation only after establishing whether the antibody epitope faces the cell exterior or interior; its orientation is not supplied here (UniProt Q13336 topology; standard IF practice). The HPA subcellular record reports plasma-membrane localisation and lists RT-4 IF/ICC images, which can guide compartment assessment without defining this antibody’s IF conditions (HPA subcellular). Include single-label and no-primary controls to check bleed-through and background (standard IF practice).
What explains diffuse brown staining in SLC14A1 paraffin sections?
Check the no-primary control first to distinguish antibody-dependent signal from detection background (standard IHC practice). The catalog image used a peroxidase-linked secondary antibody and DAB, so a peroxidase block and careful inspection for endogenous pigment are relevant general workflow checks (datasheet A04926-2; standard IHC practice). In the illustrated melanoma tissue, inspect pigmented regions before calling brown deposits SLC14A1 staining (datasheet A04926-2; standard IHC practice). If the control is clean, compare blocking, washing, and the documented 2 μg/ml primary-antibody condition on adjacent sections (datasheet A04926-2; standard IHC practice). Score defined cells and compartments rather than diffuse deposits across tissue edges or damaged areas (standard IHC practice).
How should I quantify SLC14A1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because erythrocytes are an abundant reported signal source and tissue staining includes other cell types (HPA tissue IHC). For a defined population, report the percentage of positive cells and staining intensity, or calculate an H-score from intensity-weighted percentages on a 0–300 scale (standard IHC practice). For spatial comparisons, report positive-cell density per mm² of viable tissue and record the area sampled (standard IHC practice). Normalize cell-based results to the number of evaluable cells of the specified type, keeping erythrocytes separate from neighboring tissue cells (standard IHC practice). Apply one threshold and the same sampling rule across batches, with control sections reviewed before comparison (standard IHC practice).
When is a positive SLC14A1 IHC result convincing?
A convincing result has a defined cellular location consistent with SLC14A1 membrane localisation and the cell population being evaluated (UniProt Q13336 subcellular location; standard IHC practice). Identify erythrocytes explicitly: they contain SLC14A1 protein and are the most abundant stained population in the HPA tissue profile (UniProt Q13336 tissue specificity; HPA tissue IHC). Treat isolated nuclear staining, tissue-edge deposits, and staining confined to necrotic areas as reasons to review morphology and controls (UniProt Q13336 subcellular location; standard IHC practice). Check no-primary and peroxidase controls when brown signal could reflect detection artefact (standard IHC practice). HPA rates its tissue IHC data Approved while reporting low consistency with RNA expression, so interpret discordant tissue patterns cautiously (HPA tissue IHC).
Boster reagents
Best SLC14A1 / Urea transporter 1 IHC Antibodies
A04926-2 has real IHC data from paraffin-embedded human melanoma and pancreatic cancer sections (catalog image captions). Its listed reactivity covers human, mouse and rat (catalog reactivity).
Real IHC data
Anti-UT-B/SLC14A1 Antibody ®
Cat # A04926-2
A04926-2 is listed for IHC and for human, mouse and rat reactivity (catalog applications/reactivity). Its IHC images show paraffin-embedded human melanoma and pancreatic cancer sections (A04926-2 image captions).
Which to pick: For tissue IHC, choose A04926-2 at 2–5 μg/ml (catalog IHC dilution); its image caption documents EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody on a paraffin-embedded human section, with the fixative unreported (A04926-2 image caption). No IF/ICC-validated SKU is supplied: A04926-2 has no IF application, image or dilution listed (catalog applications/IF fields). For cross-species IHC planning, A04926-2 lists human, mouse and rat reactivity, while its IHC dilution listing specifies human and rat and its shown IHC sections are human (catalog reactivity/dilution; A04926-2 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q13336 (UT1_HUMAN, Urea transporter 1).
- Human Protein Atlas. SLC14A1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. SLC14A1 subcellular location (ICC-IF): Localized to the plasma membrane..
- Human Protein Atlas. SLC14A1 antibody validation summary (2 antibodies).
- SLC14A1 prevents oncometabolite accumulation and recruits HDAC1 to transrepress oncometabolite genes in urothelial carcinoma. Theranostics 2020 — PMC7546005.
- SLC14A1 and TGF-β signaling: a feedback loop driving EMT and colorectal cancer metachronous liver metastasis. Journal of experimental & clinical cancer research : CR 2024 — PMC11282742.
- Study on the role of SLC14A1 gene in biochemical recurrence of prostate cancer. Scientific reports 2022 — PMC9579171.
- PubMed PMID:7989337 — UniProt-cited evidence.
- PubMed PMID:8573795 — UniProt-cited evidence.
- PubMed PMID:9215669 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SLC14A1 (Urea transporter 1). Expected localisation Cytoplasmic stain (HPA tissue IHC); membrane expected (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04926-2). Validated IHC antibody: A04926-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Processes in granular layer); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human melanoma tissue; fixative not specified (datasheet A04926-2); 2. Fixation: Image fixative and duration unreported (datasheet A04926-2); 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 retrieval: EDTA pH 8.0 (datasheet A04926-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04926-2); 8. Primary antibody: Rabbit anti-SLC14A1, 2-5 μg/ml (datasheet A04926-2); 9. Primary incubation: Overnight at 4 °C (datasheet A04926-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Antibody staining has low concordance with RNA (HPA tissue IHC); Isoform / epitope: Two isoforms; map extracellular vs cytoplasmic epitopes (UniProt); Regulation: Regulation not established (UniProt). Evidence: UniProt Q13336, HPA tissue IHC · Approved, HPA ICC-IF · plasma membrane., Datasheet A04926-2, PMC7546005, PMC11282742. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SLC14A1/immunohistochemistry-ihc-guide.