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SLC5A2 / Sodium/glucose cotransporter 2 · IHC design guide
Design Immunohistochemistry for SLC5A2
Plan chromogenic paraffin IHC for SLC5A2 using the membranous staining of kidney proximal-tubule microvilli as the expected pattern (HPA tissue IHC). A catalog antibody has a kidney-section IHC example at 1 μg/mL with DAB detection (datasheet A03748-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SLC5A2 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 | Proximal-tubule microvilli membrane (HPA tissue IHC) | |
| Staining pattern | Membranous staining of kidney proximal-tubule microvilli (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A03748-1) | |
| Positive control | Kidney | |
| 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 | Endogenous renal biotin may mimic SABC signal (standard IHC practice; datasheet A03748-1 detection) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope map unknown; extracellular versus cytoplasmic location matters (UniProt) |
Section 1
Recommended SLC5A2 IHC & IF Protocols
The catalog antibody has a datasheet IHC-P protocol (datasheet A03748-1); the published options below report SLC5A2 staining conditions from three PMC articles (PMC8720966; PMC7587001; PMC10599739).
| Sample | Paraffin-embedded human renal cancer tissues; fixative not specified (datasheet A03748-1) |
| Fixation | Image fixative and duration unreported (datasheet A03748-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A03748-1) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03748-1) |
| Primary antibody | Rabbit anti-SLC5A2, 0.5-1μg/ml (datasheet A03748-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03748-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03748-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SLC5A2-positive staining in proximal tubules (microvilli) of kidney (HPA tissue IHC: High). HPA tissue profile: Membranous expression in proximal tubules in kidney. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6 heat retrieval for the catalog antibody (datasheet A03748-1). The no-retrieval condition used a different antibody (PMC7587001).
Section 2
What Is the Expected SLC5A2 Staining Pattern?
SLC5A2 should appear at the apical membrane of kidney proximal tubules, where the luminal microvilli stain strongly (UniProt P31639: apical cell membrane; HPA tissue IHC: High in proximal tubule microvilli). Its 14 transmembrane segments support a membrane pattern (UniProt P31639 topology). HPA rates the kidney IHC pattern Enhanced, citing consistency between antibody staining and RNA expression (HPA tissue IHC: Enhanced).
| Strong, narrow staining along the luminal edge of kidney proximal tubules (HPA tissue IHC: High in proximal tubule microvilli). | This fits the expected apical, membranous pattern (UniProt P31639: apical cell membrane; HPA tissue IHC: membranous proximal tubule expression). In chromogenic IHC, score the luminal rim and the cells carrying it; do not require individual microvilli to be resolved (general IHC practice). |
| Predominantly nuclear, diffuse cytoplasmic, or basolateral staining in proximal tubules (relative to UniProt P31639: apical cell membrane). | A dominant signal outside the expected apical rim does not match the reported localisation (UniProt P31639: apical cell membrane; HPA tissue IHC: proximal tubule microvilli). Check whether the signal follows tissue structures consistently, then compare it with an appropriate negative control (general IHC practice). |
| Strong staining in unrelated cells, such as adipocytes or adrenal glandular cells (HPA tissue IHC: Not detected in these cells). | These cells are useful comparators, but an HPA 'Not detected' result is not proof of universal absence (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity; inspect the negative controls before assigning the signal to SLC5A2 (general IHC practice). |
| Broad colour across tissue and empty spaces, without a clear proximal tubule luminal rim (relative to HPA tissue IHC: proximal tubule microvilli). | This is poorly resolved background rather than the reported selective membrane pattern (HPA tissue IHC: membranous proximal tubule expression). Review the no-primary control, blocking and wash conditions, and detection development (general IHC practice). |
| No visible luminal signal in an adequately sampled kidney section (HPA tissue IHC: High in proximal tubule microvilli). | The expected positive compartment is missing (UniProt P31639: apical cell membrane; HPA tissue IHC: High in proximal tubule microvilli). Confirm proximal tubules are present and review the IHC workflow and positive control before interpreting other tissues as negative (general IHC practice). |
💡Expected SLC5A2 appearanceCall a section positive when proximal tubules show High, selective luminal apical membrane staining (HPA tissue IHC: High in proximal tubule microvilli; UniProt P31639: apical cell membrane); widespread nuclear, diffuse, or unrelated-cell signal is a suspect pattern that needs controls (general IHC practice).
| Anatomic compartment (HPA tissue IHC; UniProt P31639). | Use the proximal tubule luminal border as the scoring landmark: HPA reports membranous staining of microvilli, consistent with UniProt's apical membrane assignment (HPA tissue IHC; UniProt P31639: apical cell membrane). |
| Tissue comparators (HPA tissue IHC). | Kidney is the documented High comparator; testis cells in seminiferous ducts are reported Low, while the listed adipocytes and adrenal glandular cells are Not detected (HPA tissue IHC). Treat each result as a reported staining level, not a universal rule for every specimen (HPA tissue IHC). |
| IHC validation (HPA antibody record; HPA tissue IHC). | HPA lists HPA041603 as IHC Enhanced and describes the tissue pattern as consistent with RNA expression (HPA antibody record: IHC Enhanced; HPA tissue IHC: Enhanced). That supports the reported pattern; a new antibody or local workflow still needs its own controls (general IHC practice). |
| Membrane topology and unknown epitope (UniProt P31639 topology). | SLC5A2 has 14 transmembrane segments and extracellular and cytoplasmic regions (UniProt P31639 topology). No antibody epitope is supplied, so topology alone cannot justify a specific antigen retrieval or permeabilisation setting (UniProt P31639 topology; supplied antibody record). |
| Isoforms and processing (UniProt P31639). | UniProt lists 2 isoforms and a chain spanning residues 1–672, with no annotated signal peptide or propeptide (UniProt P31639). Without an epitope or isoform-specific staining record, the slide cannot distinguish isoforms or support a processing explanation (UniProt P31639; HPA tissue IHC). |
| IF/ICC? (HPA subcellular; HPA antibody record). | HPA gives a general Membrane summary but no main location or ICC-IF image cell lines, and the supplied antibody record has no ICC validation entry (HPA subcellular; HPA antibody record). The IHC proximal tubule pattern does not establish an IF/ICC result (HPA tissue IHC; HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| Kidney positive control has no apical rim (HPA tissue IHC: High in proximal tubule microvilli). | The section may lack identifiable proximal tubules, or the staining workflow may have failed (general IHC practice); the image alone cannot identify which. | Confirm proximal tubules on the counterstained section, then check antibody application, retrieval, detection and a working positive control (general IHC practice). Avoid calling other tissue negative until the control works. |
| Signal is chiefly nuclear or basolateral (relative to UniProt P31639: apical cell membrane). | The pattern conflicts with the reported apical localisation (UniProt P31639; HPA tissue IHC). Its cause cannot be assigned from localisation alone. | Recheck compartment boundaries and compare no-primary and tissue controls; score only a reproducible apical proximal tubule pattern as expected SLC5A2 staining (general IHC practice; HPA tissue IHC). |
| Adipocytes or adrenal glandular cells stain strongly (HPA tissue IHC: Not detected in these cells). | Cross-reactivity or endogenous detection activity is possible (general IHC practice); HPA's negative report alone cannot identify the cause (HPA tissue IHC). | Compare a no-primary control and an appropriate negative tissue, then review blocking and detection chemistry (general IHC practice). Keep the unexpected cells separate from the kidney positive score (HPA tissue IHC). |
| Diffuse deposit obscures the luminal edge (relative to HPA tissue IHC: proximal tubule microvilli). | Nonspecific background or overdeveloped chromogen can obscure membrane detail (general IHC practice). | Review the no-primary control, washes, blocking, primary concentration and development time, changing one workflow variable at a time (general IHC practice). Reassess whether a distinct apical rim remains (HPA tissue IHC). |
| Weak testis staining is treated as a failed negative control (HPA tissue IHC: Low in cells of seminiferous ducts). | Testis is reported Low rather than Not detected in the supplied tissue profile (HPA tissue IHC). | Use a reported Not detected cell type for a tissue comparison and keep the kidney proximal tubule as the positive reference (HPA tissue IHC). Interpret weak testis staining against local controls (general IHC practice). |
| An IF/ICC image is used to claim a validated cell-line pattern (HPA subcellular: no ICC-IF image cell lines). | The supplied HPA summary says Membrane but provides no main location or ICC-IF image cell lines; HPA041603 has no ICC validation entry here (HPA subcellular; HPA antibody record). | Report the IF/ICC observation as an observation requiring its own controls (general IF practice). Use the IHC kidney result only as tissue-pattern context (HPA tissue IHC). |
Experiment evidence
IHC & IF images showing SLC5A2 staining
A03748-1 · IHCIHC analysis of SLC5A2 using anti-SLC5A2 antibody (A03748-1). SLC5A2 was detected in paraffin-embedded section of human renal cancer tissues
A03748-1 · IHCIHC analysis of SLC5A2 using anti-SLC5A2 antibody (A03748-1). SLC5A2 was detected in paraffin-embedded section of mouse kidney tissues. Heat
A03748 · IHCImmunohistochemical analysis of paraffin-embedded human Colon cancer. 1, Antibody was diluted at 1:200(4° overnight). 2, Tris-EDTA,pH9.0 was
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000140675-SLC5A2/tissue/cerebral+cortex#imid_18403587
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000140675-SLC5A2/tissue/colon#imid_18399177
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000140675-SLC5A2/tissue/liver#imid_18399194
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000140675-SLC5A2/tissue/kidney#imid_18399179
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SLC5A2 IHC & IF
🧪Run kidney first: proximal tubule microvilli should show strong apical staining (HPA: High in proximal tubules [microvilli]; UniProt P31639: apical cell membrane). Use adipose tissue as the negative tissue, where adipocytes are not detected; neighboring non-proximal renal cells should lack the distinct apical microvillar pattern, although their antigen-negative status is not established by the supplied HPA row (HPA: Not detected in adipocytes; HPA: High in proximal tubules [microvilli]).
Positive control tissue: Kidney (Proximal tubules (microvilli), 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 SLC5A2; derive a cell-line control from the positive tissue's cell type (Proximal tubules (microvilli)) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; a rabbit IgG isotype control matched to the primary antibody’s clonality and concentration; and SLC5A2 knockout material or immunizing-peptide competition if available (selected-SKU IHC caption: rabbit primary antibody). For kidney sections, quench endogenous peroxidase and check for endogenous biotin when using the caption’s biotinylated secondary, streptavidin complex and DAB detection (selected-SKU IHC caption: biotinylated secondary, SABC and DAB).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the exact selected-SKU tissue-IHC caption leaves the fixative unreported (selected-SKU IHC caption: fixative not stated). The caption uses heat retrieval in citrate buffer at pH 6 for 20 minutes, but does not establish whether retrieval is required under other conditions (selected-SKU IHC caption: citrate retrieval). Frozen sections and IF cannot be judged easier from the supplied evidence; in kidney, assess the apical microvillar pattern against background in adjacent structures (HPA: High in proximal tubules [microvilli]; HPA subcellular: no ICC-IF cell-line images).
HPA tissue IHC evidence for SLC5A2
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 |
|---|---|---|---|---|
| Kidney | Proximal tubules (microvilli) | High | 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 SLC5A2 IHC Tips
Troubleshoot SLC5A2 staining in paraffin sections by checking retrieval, membrane localisation, controls and scoring against the expected proximal tubule pattern (HPA tissue IHC; UniProt P31639).
What retrieval conditions should I try when SLC5A2 staining is weak?
Start with heat-mediated citrate retrieval at pH 6 for 20 minutes (caption A03748-1). Keep heating and cooling consistent across sections, then compare staining at the proximal tubule brush border against a section processed in the same run (general IHC practice; HPA tissue IHC). The published product image used a paraffin section of human renal cancer tissue, but its caption does not state the fixative (caption A03748-1). If staining remains weak, test a different retrieval condition on adjacent sections as a documented fallback, while checking whether increased signal also raises nonspecific staining (general IHC practice).
How should I assess whether fixation is affecting SLC5A2 staining?
Target-specific fixation sensitivity is unknown from the supplied evidence; the selected product caption identifies a paraffin section but does not report its fixative (caption A03748-1). Record the fixative and fixation duration for each specimen, and compare sections processed with the same retrieval and detection conditions (general IHC practice). Include a kidney section with proximal tubules as a reference for the expected membranous pattern (HPA tissue IHC). If signal differs between specimens, inspect tissue preservation and processing history before attributing the difference to SLC5A2 abundance (general IHC practice). Do not infer a fixation effect from the protein’s membrane topology or HPA staining profile (UniProt P31639 topology; HPA tissue IHC).
Which staining pattern supports specific SLC5A2 detection?
Look for membranous staining along proximal tubule microvilli in kidney, with the apical surface as the expected compartment (HPA tissue IHC; UniProt P31639 subcellular location). Compare the luminal edge with lateral membranes, cytoplasm and nearby structures on the same section rather than judging intensity alone (general IHC practice). SLC5A2 has 14 annotated transmembrane segments, so a diffuse nuclear pattern requires particularly careful validation (UniProt P31639 topology). The selected product image shows staining in a human renal cancer section, which does not by itself establish the expected distribution in every tumour area (caption A03748-1).
How can epitope location affect interpretation of SLC5A2 IHC?
Confirm the antibody’s immunogen or mapped epitope before assigning a negative result to loss of SLC5A2; that information is absent from the supplied product caption (caption A03748-1). UniProt lists 2 isoforms and a protein with 14 transmembrane segments, extracellular and cytoplasmic regions, and one annotated glycosylation site at residue 250 (UniProt P31639). Those features make epitope accessibility a plausible experimental variable, but the supplied evidence does not identify which region this antibody recognises (UniProt P31639 topology; caption A03748-1). Compare retrieval conditions and a positive kidney reference section before interpreting an absent apical signal (general IHC practice; HPA tissue IHC).
How should I investigate an SLC5A2 IF result alongside chromogenic IHC?
Use the IHC result as an anatomical reference: SLC5A2 is expected at the apical membrane of kidney proximal tubules (HPA tissue IHC; UniProt P31639 subcellular location). In a separate IF experiment, multiplex with a validated proximal tubule marker and assess whether both signals occupy the expected cells and membrane region (general IF practice; HPA tissue IHC). Choose fluorophores and imaging channels after measuring tissue autofluorescence, and include single-stain controls for channel bleed-through (general IF practice). Set permeabilisation with the antibody epitope in mind: cytoplasmic epitopes require access through the membrane, while excessive permeabilisation can complicate membrane-pattern assessment (general IF practice; UniProt P31639 topology).
What should I check when the chromogenic signal obscures proximal tubules?
First compare stained sections with a no-primary control and inspect whether colour follows tissue edges, damaged regions or the expected apical membrane (general IHC practice; HPA tissue IHC). The selected product image used 10% goat serum blocking, 1 μg/ml primary antibody overnight at 4°C, biotinylated secondary antibody and DAB development (caption A03748-1). Optimise primary concentration and development time on adjacent sections if staining is widespread, while holding retrieval constant (general IHC practice). Include a peroxidase block and appropriate detection controls to assess chromogen background; these are general IHC workflow steps, not evidence of SLC5A2 specificity (general IHC practice).
How should I quantify SLC5A2 staining across kidney sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define proximal tubules morphologically and score the apical membranous compartment, where the reference pattern is strongest (HPA tissue IHC; UniProt P31639 subcellular location). Report the percentage of positive proximal tubules or cells together with membrane intensity; an H-score can combine percentage and intensity if its scoring bins are defined in advance (general IHC practice). Normalise counts to the total evaluable proximal tubules or proximal tubule area, rather than all tissue area when composition differs between sections (general IHC practice). Exclude folds, necrosis and poorly preserved regions using the same rules across specimens, and retain representative images for review (general IHC practice).
How can I distinguish true SLC5A2 signal from staining artefact?
Treat apical staining in kidney proximal tubules as the principal positive pattern, supported by the reported high microvillar signal and apical membrane localisation (HPA tissue IHC; UniProt P31639 subcellular location). Reassess signal concentrated in nuclei, unrelated cell populations, section edges or necrotic regions before calling it SLC5A2 expression (general IHC practice; HPA tissue IHC). Compare the result with a no-primary control and inspect whether endogenous enzyme activity contributes to DAB colour (general IHC practice). HPA reports SLC5A2 as kidney enriched and absent from several listed tissues, but an unexpected result still needs control and morphology checks rather than dismissal by tissue name alone (HPA tissue IHC).
Boster reagents
Best SLC5A2 / Sodium/glucose cotransporter 2 IHC Antibodies
Both antibodies have IHC images from human paraffin sections; A03748-1 also has mouse and rat kidney images (catalog IHC captions). ICC is listed for A03748-1, but no IF image is supplied (catalog applications and images).
Real IHC data
Anti-SGLT2/SLC5A2 Antibody ®
Cat # A03748-1
Real IHC data
Anti-SLC5A2/Sglt2 Antibody
Cat # A03748
A03748 will render with its human colon cancer paraffin-section IHC image; its listed reactivity is human, mouse and rat (A03748 image caption and reactivity). A03748-1 will render with its human renal cancer paraffin-section IHC image; additional captions show mouse and rat kidney sections (A03748-1 IHC captions).
Which to pick: For tissue IHC, choose A03748-1 when kidney evidence or cross-species images matter; its captions show human renal cancer and mouse and rat kidney paraffin sections, while the polyclonal A03748 has a human colon cancer paraffin-section image (catalog IHC captions; A03748 dilution_raw). For IF/ICC work, A03748-1 lists ICC at 0.5–1 μg/ml, but supplies no IF image; A03748 does not list ICC or IF (catalog applications, dilutions and images). The IHC captions specify paraffin sections but do not report the fixative for either SKU (catalog IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P31639 (SC5A2_HUMAN, Sodium/glucose cotransporter 2).
- Human Protein Atlas. SLC5A2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SLC5A2 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. SLC5A2 antibody validation summary (1 antibodies).
- High-Salt Attenuates the Efficacy of Dapagliflozin in Tubular Protection by Impairing Fatty Acid Metabolism in Diabetic Kidney Disease. Frontiers in pharmacology 2021 — PMC8720966.
- Sodium/glucose cotransporter 2 is expressed in choroid plexus epithelial cells and ependymal cells in human and mouse brains. Neuropathology : official journal of the Japanese Society of Neuropathology 2020 — PMC7587001.
- Analysis of potential biomarkers for diabetic kidney disease based on single-cell RNA-sequencing integrated with a single-cell sequencing assay for transposase-accessible chromatin. Aging 2023 — PMC10599739.
- Bone from Healthy Individuals and Patients with CKD Expresses the Sodium-Glucose Co-transporter-2 (SGLT2). Calcified tissue international 2026 — PMC12982296.
- PubMed PMID:1415574 — UniProt-cited evidence.
- PubMed PMID:15616553 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SLC5A2 (Sodium/glucose cotransporter 2). Expected localisation Proximal-tubule microvilli membrane (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A03748-1). Validated IHC antibody: A03748-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Proximal tubules (microvilli)); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human renal cancer tissues; fixative not specified (datasheet A03748-1); 2. Fixation: Image fixative and duration unreported (datasheet A03748-1); 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: Citrate pH 6 , 20 min (datasheet A03748-1); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03748-1); 8. Primary antibody: Rabbit anti-SLC5A2, 0.5-1μg/ml (datasheet A03748-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03748-1). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Endogenous renal biotin may mimic SABC signal (standard IHC practice; datasheet A03748-1 detection); Isoform / epitope: 2 isoforms; epitope map unknown; extracellular versus cytoplasmic location matters (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt P31639, HPA tissue IHC · Enhanced, HPA ICC-IF · Membrane, Datasheet A03748-1, PMC8720966, PMC7587001. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SLC5A2/immunohistochemistry-ihc-guide.