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SLC5A5 / Sodium/iodide cotransporter · IHC design guide
Design Immunohistochemistry for SLC5A5
Plan chromogenic IHC for SLC5A5 using the IHC-validated antibody A04465-1 (datasheet). Compare membranous and cytoplasmic staining in glandular cells, with thyroid gland and stomach as high-staining reference tissues (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SLC5A5 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 | Glandular cells: membranous and cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04465-1) | |
| Positive control | Choroid plexus+3 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 A04465-1) | |
| Caveat | Presumed off-target staining requires careful interpretation (HPA tissue IHC) | |
| Regulation | Expression is reduced in tumors (UniProt) | |
| Isoform / epitope | No annotated isoforms; epitope side matters across 13 spans (UniProt) |
Section 1
Recommended SLC5A5 IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet: A04465-1). The published protocols below report SLC5A5 staining in paraffin sections and frozen sections (PMC5754505; PMC12155144; PMC6735285).
| Sample | Paraffin-embedded rat stomach tissue; fixative not specified (datasheet A04465-1) |
| Fixation | Image fixative and duration unreported (datasheet A04465-1); 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 A04465-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04465-1) |
| Primary antibody | Rabbit anti-SLC5A5, 1:50 recommended; image 1:100 (datasheet A04465-1) |
| Primary incubation | Overnight at 4 °C (datasheet A04465-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04465-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SLC5A5-positive staining in ependymal cells of choroid plexus (HPA tissue IHC: High). HPA tissue profile: Selective cytoplasmic and membranous expression, most abundant in choroid plexus, salivary ducts, thyroid gland and stomach. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet: A04465-1). Citrate pH 6.0 is a published alternative for a different antibody (PMC5754505).
Section 2
What Is the Expected SLC5A5 Staining Pattern?
SLC5A5 is a 13-transmembrane protein assigned to the cell membrane and cytoplasm (UniProt Q92911 topology and subcellular location). In paraffin-section IHC, expect selective membranous and cytoplasmic staining in glandular cells of thyroid and stomach, ependymal cells of choroid plexus, and glandular cells of salivary gland (HPA tissue IHC). HPA rates its tissue staining reliability “Enhanced” and notes that presumed off-target staining was disregarded (HPA tissue IHC).
| Distinct cell-boundary staining, with some cytoplasmic signal, in the expected cells. | This matches the reported membrane and cytoplasm locations (UniProt Q92911 subcellular location) and HPA’s selective membranous and cytoplasmic tissue pattern (HPA tissue IHC). Judge the result by both compartment and cell type; chromogen colour alone does not establish specificity (general IHC practice). |
| Predominantly nuclear staining, or staining detached from recognisable cell boundaries and cytoplasm. | A predominantly nuclear pattern is unsupported by the supplied localisation records (UniProt Q92911 subcellular location; HPA tissue IHC). Treat it as suspect and compare with controls and tissue morphology before interpreting it as SLC5A5 (general IHC practice). |
| Strong staining in an unexpected cell population, including cells listed as not detected. | HPA reports no detection in adipocytes of adipose tissue or hematopoietic cells of bone marrow, among other listed populations (HPA tissue IHC). Unexpected signal can reflect cross-reactivity or detection-system activity; check a reagent-only control and confirm which cells carry the stain (general IHC practice). |
| Diffuse colour across tissue or in areas without intact cells. | This does not resemble HPA’s selective cell-associated pattern (HPA tissue IHC). Background from the staining workflow can obscure genuine membrane signal; inspect the negative control and review blocking, washes and detection reagents (general IHC practice). |
| No staining in an otherwise interpretable thyroid, stomach or choroid plexus section. | HPA reports High staining in thyroid and stomach glandular cells and choroid plexus ependymal cells (HPA tissue IHC). Absence in the expected cells calls for a control and workflow check before concluding that the specimen lacks SLC5A5 (general IHC practice). |
💡Expected SLC5A5 appearanceCall a positive result when the expected cells show selective membranous staining with possible cytoplasmic signal, typically High in thyroid or stomach glandular cells and choroid plexus ependymal cells; widespread nuclear or cell-free colour is suspect (HPA tissue IHC; UniProt Q92911 subcellular location; general IHC practice).
| Cell compartment and topology | UniProt assigns SLC5A5 to membrane and cytoplasm and annotates 13 transmembrane segments (UniProt Q92911 topology and subcellular location). Use cell-associated membrane signal as the localisation anchor; topology alone does not specify which membrane face an IHC antibody recognises (general IHC practice). |
| Choice of positive and comparison tissue | HPA records High staining in choroid plexus ependymal cells and thyroid and stomach glandular cells, versus Medium in salivary gland glandular cells (HPA tissue IHC). Compare the named cells, since an overall tissue score can hide an incorrect cell pattern (general IHC practice). |
| Strength of pattern evidence | HPA rates tissue staining “Enhanced” for consistency with RNA expression and records IHC “Enhanced” for 2 antibodies, HPA049055 and CAB022364 (HPA tissue IHC; HPA antibodies). This supports the reported pattern, while HPA also notes that presumed off-target binding was disregarded (HPA tissue IHC). |
| Scope of IF/ICC evidence | HPA’s subcellular summary says “Membrane,” but provides no main-location assignment or ICC-IF image-bearing cell lines; the listed antibodies have no ICC status (HPA subcellular; HPA antibodies). That summary can inform an IF/ICC localisation check, but it does not supply an IF/ICC staining result for this guide (HPA subcellular; HPA antibodies). |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive-control section has no visible staining. | Thyroid and stomach glandular cells and choroid plexus ependymal cells are reported High by HPA, so absent signal may reflect a staining-run problem or the cells present in the section (HPA tissue IHC; general IHC practice). | Confirm that the named cells are present, then review the antibody, antigen-retrieval and detection steps against the validated IHC workflow; use a same-run positive control (general IHC practice). |
| The slide is brown throughout, including areas without the expected cells. | Diffuse colour is inconsistent with HPA’s selective cell-associated pattern; incomplete blocking, washing or control of endogenous detection activity can contribute to IHC background (HPA tissue IHC; general IHC practice). | Inspect the primary-antibody-omission control, review blocking and wash steps, and check the detection system’s endogenous-activity control before scoring cells (general IHC practice). |
| Nuclei stain more strongly than cell borders. | The supplied sources place SLC5A5 at the membrane and in cytoplasm, and HPA describes membranous and cytoplasmic tissue staining (UniProt Q92911 subcellular location; HPA tissue IHC). A nuclear-dominant result is therefore unsupported by this evidence. | Check the counterstain and detection controls, then reassess localisation in morphologically intact cells; do not score nuclear-only colour as a supported SLC5A5 pattern (general IHC practice; UniProt Q92911 subcellular location). |
| A listed negative cell type stains strongly. | HPA lists adipose-tissue adipocytes and bone-marrow hematopoietic cells as not detected (HPA tissue IHC). Signal there may reflect nonspecific binding or detection-system activity, though a single unexpected slide does not identify its cause (general IHC practice). | Verify cell identity and compare with a reagent-only control; repeat the interpretation against a same-run expected-positive tissue and the HPA cell-specific pattern (general IHC practice; HPA tissue IHC). |
| Cytoplasmic staining appears without a clear cell-border signal. | Cytoplasm is a reported location, but HPA describes a selective cytoplasmic and membranous tissue pattern (UniProt Q92911 subcellular location; HPA tissue IHC). Cytoplasmic colour alone gives weaker localisation support than a matching cell and membrane pattern (general IHC practice). | Confirm that staining is confined to an HPA-reported positive cell population, inspect a negative control, and score the cytoplasmic and membrane components separately (HPA tissue IHC; general IHC practice). |
| Q: Can the IHC result be used as an IF/ICC protocol or validation result? | HPA lists a membrane subcellular summary but no ICC-IF image-bearing cell lines or ICC validation status for the listed antibodies (HPA subcellular; HPA antibodies). | A: Use the separate IF/ICC guide for that application; describe this section’s cell and tissue expectations as IHC evidence, and do not transfer IHC antibody validation to IF/ICC (HPA tissue IHC; HPA antibodies; general IHC practice). |
Experiment evidence
IHC & IF images showing SLC5A5 staining
A04465-1 · IHCIHC analysis of SLC5A5 using anti-SLC5A5 antibody (A04465-1). SLC5A5 was detected in a paraffin-embedded section of rat stomach tissue. Heat
A04465-1 · IHCIHC analysis of SLC5A5 using anti-SLC5A5 antibody (A04465-1). SLC5A5 was detected in a paraffin-embedded section of mouse stomach tissue. He
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000105641-SLC5A5/tissue/cerebral+cortex#imid_21206539
HPA · Choroid plexusHuman Protein Atlas IHC, Choroid plexus. Source: https://www.proteinatlas.org/ENSG00000105641-SLC5A5/tissue/choroid+plexus#imid_4066
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000105641-SLC5A5/tissue/colon#imid_21206473
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000105641-SLC5A5/tissue/liver#imid_21206482
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SLC5A5 IHC & IF
🧪Run stomach first: glandular cells should stain (HPA: High in stomach glandular cells; selected A04465-1 caption: staining in a paraffin section of rat stomach). Use adipose tissue as a negative control, with adipocytes at background (HPA: Not detected in adipocytes); nearby non-glandular cells on the positive slide should also remain at background (standard IHC internal-control practice).
Positive control tissue: Choroid plexus (Ependymal cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for SLC5A5; derive a cell-line control from the positive tissue's cell type (Ependymal cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; a concentration-matched rabbit IgG isotype control; and, if available, SLC5A5 knockout tissue or a validated peptide-block control (selected A04465-1 caption: rabbit primary antibody; standard IHC specificity controls). Quench endogenous peroxidase and assess background before scoring stomach staining with HRP/DAB (selected A04465-1 caption: HRP/DAB detection; standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A04465-1 tissue-IHC caption does not state the fixative. The caption documents heat retrieval in EDTA at pH 8.0 for a paraffin section, so retrieval is a practical starting point, but its necessity has not been established (selected A04465-1 caption). The supplied evidence does not establish that frozen sections or IF are easier; for stomach HRP/DAB, check endogenous peroxidase background (selected A04465-1 caption: HRP/DAB; standard IHC practice).
HPA tissue IHC evidence for SLC5A5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). 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 |
|---|---|---|---|---|
| Choroid plexus | Ependymal cells | High | Protein (IHC) | HPA → |
| Stomach | Glandular cells | High | Protein (IHC) | HPA → |
| Thyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SLC5A5 IHC Tips
Troubleshoot SLC5A5 staining in paraffin sections by checking retrieval, cell type, membrane localisation and assay controls before interpreting chromogenic signal.
What retrieval should I try first if SLC5A5 staining is weak?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A04465-1). The selected tissue image used this retrieval before overnight incubation at 4°C with antibody diluted 1:100; its caption does not specify a heating time (datasheet A04465-1). Keep section thickness, heating duration and cooling conditions consistent across test slides, then compare signal in stomach glandular cells with background in adjacent structures (HPA tissue IHC: high in stomach glandular cells; standard IHC practice). If staining remains weak, test a different retrieval condition on matched sections as a fallback, while retaining the documented EDTA condition as the reference (standard IHC practice).
Could fixation explain weak or uneven SLC5A5 staining?
The selected paraffin-section caption does not state a fixative, so target-specific SLC5A5 sensitivity to fixation is unknown (datasheet A04465-1). Record each specimen's fixative and fixation interval, and compare sections processed together before attributing a signal difference to the antibody (standard IHC practice). If staining varies across a block, inspect tissue preservation and repeat staining on a matched section with the documented EDTA pH 8.0 retrieval condition (datasheet A04465-1; standard IHC practice). Include a stomach or thyroid gland section as a positive comparator, since glandular cells in both are reported to stain strongly (HPA tissue IHC: high in stomach and thyroid glandular cells).
How should I assess membrane versus cytoplasmic SLC5A5 staining?
Assess membrane-associated signal separately from cytoplasmic signal because SLC5A5 is annotated in both compartments (UniProt Q92911: cell membrane and cytoplasm). Its 13 transmembrane segments make cell-boundary staining biologically plausible, while a diffuse cytoplasmic pattern needs comparison with controls and tissue morphology (UniProt Q92911 topology; standard IHC practice). HPA describes selective cytoplasmic and membranous staining, most abundant in choroid plexus, salivary ducts, thyroid gland and stomach (HPA tissue IHC). Score glandular cells in stomach or thyroid gland independently from surrounding cells, and document whether signal follows intact cell outlines or pools in damaged regions (HPA tissue IHC: high in stomach and thyroid glandular cells; standard IHC practice).
How does epitope position affect interpretation of SLC5A5 staining?
Check the antibody's stated immunogen or epitope before interpreting a negative section; the selected caption gives no epitope position (datasheet A04465-1). UniProt lists 13 transmembrane segments, an extracellular region at residues 466–525, and a cytoplasmic tail at 551–643 (UniProt Q92911 topology). Glycosylation sites at 489 and 502 lie in that extracellular region, and phosphoserine 556 lies in the cytoplasmic tail (UniProt Q92911 topology and PTMs). If two antibodies disagree, compare their documented epitopes and retrieval requirements on matched sections before assigning a biological difference; UniProt lists 0 annotated isoforms (UniProt Q92911 isoforms; standard IHC practice).
How can I check a disputed IHC pattern by immunofluorescence?
Use IF as a separate validation experiment and multiplex SLC5A5 with an independently validated marker of the expected glandular cell population (HPA tissue IHC: high in stomach and thyroid glandular cells; standard IF practice). Choose fluorophores after examining unstained tissue in each channel, placing the target signal in a channel with less tissue autofluorescence (standard IF practice). Set permeabilisation according to the antibody's documented epitope: an extracellular epitope may be accessible without it, whereas the cytoplasmic 551–643 tail requires membrane access (UniProt Q92911 topology; standard IF practice). Compare single-stain and secondary-only controls with the multiplex image before treating overlap as corroboration of the chromogenic pattern (standard IF practice).
What should I check when DAB staining appears widespread?
Run a no-primary control to assess secondary reagent, endogenous peroxidase and chromogen background in the chromogenic workflow (standard IHC practice). The selected image used a peroxidase-conjugated secondary and DAB, with 10% goat serum blocking and primary antibody at 1:100 overnight at 4°C (datasheet A04465-1). Apply an appropriate peroxidase block, then compare matched sections with and without primary antibody before changing its dilution or incubation (standard IHC practice). Treat broad staining outside the reported selective glandular and membranous or cytoplasmic pattern cautiously, particularly because HPA reports presumed off-target binding in its tissue assessment (HPA tissue IHC).
How should I quantify SLC5A5 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; HPA reports strong staining in stomach and thyroid glandular cells and both membranous and cytoplasmic expression (HPA tissue IHC). For each compartment, record the percentage of positive glandular cells and an intensity-based H-score using a prespecified threshold applied consistently across slides (standard IHC practice). Normalise counts to the number of evaluable glandular cells, or normalise positive-cell density to evaluable tissue area in mm²; exclude folds, necrosis and section edges (standard IHC practice). Keep retrieval, chromogen development and image acquisition comparable across specimens, and report membrane and cytoplasmic scores separately (standard IHC practice; UniProt Q92911: cell membrane and cytoplasm).
When is an apparent SLC5A5-positive cell likely an artefact?
Give most weight to reproducible staining in expected cells and compartments: HPA reports high glandular-cell staining in stomach and thyroid gland and selective cytoplasmic and membranous expression (HPA tissue IHC). A signal restricted to torn edges, necrotic regions or areas with poor morphology warrants repeat assessment on an intact section (standard IHC practice). Diffuse staining of unrelated cells or signal in a no-primary control raises concern for nonspecific binding or endogenous enzyme activity, particularly with peroxidase and DAB detection (datasheet A04465-1; standard IHC practice). Compare a matched positive section and negative control before calling a weak cytoplasmic signal positive; HPA notes presumed off-target binding in its assessment (HPA tissue IHC).
Boster reagents
Best SLC5A5 / Sodium/iodide cotransporter IHC Antibodies
A04465-1 has real IHC images of paraffin-embedded human, mouse, and rat stomach (catalog image captions); no IF image is provided (catalog IF images).
Real IHC data
Anti-SLC5A5 Antibody
Cat # A04465-1
A04465-1 is listed for IHC and human, mouse, and rat reactivity (catalog applications and reactivity). Its own images show paraffin-embedded stomach IHC in all three species with EDTA retrieval at pH 8.0 (catalog image captions).
Which to pick: For tissue IHC, choose A04465-1: this rabbit polyclonal antibody is listed for IHC, and its own images show staining in paraffin-embedded stomach sections (catalog host, dilution data, applications, and image captions). For IF/ICC, no SKU in this payload has a listed IF/ICC application or IF image, so there is no supported pick (catalog applications and IF images). For cross-species IHC, A04465-1 lists human, mouse, and rat reactivity and shows stomach IHC in each; its image captions do not report the fixative (catalog reactivity and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q92911 (SC5A5_HUMAN, Sodium/iodide cotransporter).
- Human Protein Atlas. SLC5A5 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SLC5A5 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. SLC5A5 antibody validation summary (2 antibodies).
- NIS expression in thyroid tumors, relation with prognosis clinicopathological and molecular features. Endocrine connections 2018 — PMC5754505.
- Potassium Iodide Induces Apoptosis in Salivary Gland Cancer Cells. International journal of molecular sciences 2025 — PMC12155144.
- IGF2BP2 Drives Thyroid Cancer Dedifferentiation Through m6A-Dependent STAT1 mRNA Destabilization. International journal of biological sciences 2026 — PMC12780945.
- Enhancement of (211)At Uptake via the Sodium Iodide Symporter by the Addition of Ascorbic Acid in Targeted α-Therapy of Thyroid Cancer. Journal of nuclear medicine : official publication, Society of Nuclear Medicine 2019 — PMC6735285.
- PubMed PMID:8806637 — UniProt-cited evidence.
- PubMed PMID:9329364 — UniProt-cited evidence.
- PubMed PMID:15057824 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SLC5A5 (Sodium/iodide cotransporter). Expected localisation Membranous and cytoplasmic staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04465-1). Validated IHC antibody: A04465-1 at 1:50 recommended; image 1:100 (the product's own IHC validation image is shown). Controls (HPA): positive Choroid plexus (Ependymal cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat stomach tissue; fixative not specified (datasheet A04465-1); 2. Fixation: Image fixative and duration unreported (datasheet A04465-1); 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 A04465-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04465-1); 8. Primary antibody: Rabbit anti-SLC5A5, 1:50 recommended; image 1:100 (datasheet A04465-1); 9. Primary incubation: Overnight at 4 °C (datasheet A04465-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04465-1); Caveat: Presumed off-target staining requires careful interpretation (HPA tissue IHC); Isoform / epitope: No annotated isoforms; epitope side matters across 13 spans (UniProt); Regulation: Expression is reduced in tumors (UniProt). Evidence: UniProt Q92911, HPA tissue IHC · Enhanced, HPA ICC-IF · Membrane, Datasheet A04465-1, PMC5754505, PMC12155144. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SLC5A5/immunohistochemistry-ihc-guide.