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SLC22A12 / Solute carrier family 22 member 12 · IHC design guide
Design Immunohistochemistry for SLC22A12
Plan chromogenic IHC on paraffin kidney sections with the IHC-validated antibody A02515-2, starting at the listed 1:50 dilution (datasheet A02515-2). Assess proximal-tubule microvillar membrane staining and interpret unexpected staining cautiously because presumed off-target binding was observed (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SLC22A12 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 microvillar membrane (HPA tissue IHC) | |
| Staining pattern | Membranous staining in renal proximal-tubule microvilli (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02515-2) | |
| 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 sections. (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target binding may mimic positive staining (HPA tissue IHC) | |
| Regulation | No specific regulator reported (UniProt) | |
| Isoform / epitope | 4 isoforms; assess epitope position and loop orientation (UniProt) |
Section 1
Recommended SLC22A12 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A02515-2) is accompanied by published renal tissue IHC protocols (PMC8262335; PMC7771643).
| Sample | Paraffin-embedded human skin cancer tissue; fixative not specified (datasheet A02515-2) |
| Fixation | Image fixative and duration unreported (datasheet A02515-2); 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 A02515-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02515-2) |
| Primary antibody | Rabbit anti-SLC22A12, 1:50 recommended; image 1:100 (datasheet A02515-2) |
| Primary incubation | Overnight at 4 °C (datasheet A02515-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02515-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SLC22A12-positive staining in proximal tubules (microvilli) of kidney (HPA tissue IHC: Medium). HPA tissue profile: Distinct membranous expression in renal proximal tubules. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet: A02515-2). The mouse kidney protocol used citrate pH 6.0 (PMC7771643).
Section 2
What Is the Expected SLC22A12 Staining Pattern?
SLC22A12 is an apical membrane transporter with 12 transmembrane segments (UniProt Q96S37 topology). In kidney sections, expect membranous staining of proximal tubule microvilli; HPA reports a medium staining level and rates its tissue IHC evidence Enhanced (HPA tissue IHC). UniProt also places the protein in renal cortical proximal tubule epithelial cells (UniProt Q96S37 tissue specificity).
| A defined luminal rim outlines proximal tubules, with medium staining (HPA tissue IHC). | This fits the reported microvillar pattern and apical membrane location (HPA tissue IHC; UniProt Q96S37 subcellular location). Judge localisation as well as intensity: a brown tubular profile alone does not establish that the signal lies at the luminal surface (general IHC practice). |
| Staining chiefly fills proximal tubule cytoplasm or outlines the basolateral edge. | That compartment differs from the expected apical membrane pattern (UniProt Q96S37 subcellular location; HPA tissue IHC). Treat it as unconfirmed until the luminal border, counterstained morphology, and background controls are reviewed (general IHC practice). |
| Strong staining appears in cells outside proximal tubules or in an HPA listed negative tissue. | HPA reports proximal tubule microvilli as positive and, for example, adrenal glandular cells as not detected (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity; check a reagent omission control before assigning biological expression (general IHC practice). |
| Colour spreads across the section without a clear luminal rim. | Diffuse background cannot support the reported distinct membranous pattern (HPA tissue IHC). Compare a no primary antibody control, inspect washes and detection reagents, and score only signal that remains anatomically resolved (general IHC practice). |
| No convincing signal appears in identifiable renal proximal tubules. | The expected positive compartment is absent (HPA tissue IHC). First verify tubule identity and the IHC staining run; then review antigen retrieval, antibody dilution, and detection conditions using the antibody's own instructions (general IHC practice). |
💡Expected SLC22A12 appearanceCall positive a distinct, medium membranous rim at proximal tubule microvilli; diffuse cytoplasmic colour or staining in unrelated cells is unconfirmed (HPA tissue IHC; UniProt Q96S37 subcellular location).
| Membrane topology | SLC22A12 has 12 transmembrane segments and an apical membrane location (UniProt Q96S37 topology and subcellular location). Interpret the chromogenic result against the luminal border of intact proximal tubules, where HPA describes microvillar staining (HPA tissue IHC). |
| Tissue evidence and antibody confidence | Kidney proximal tubule microvilli show medium staining; HPA rates tissue IHC Enhanced but notes medium agreement with RNA and presumed off target binding that was disregarded (HPA tissue IHC). Use that caveat when judging unexpected staining. |
| Isoforms and modifications | UniProt lists 4 isoforms and glycosylation sites at residues 56 and 102 (UniProt Q96S37). The supplied evidence does not map the antibody epitope to an isoform or establish how these features affect this IHC stain. |
| IF/ICC Q&A: what location should an IF image show? | Expect membrane localisation from the HPA subcellular summary and apical localisation from UniProt (HPA subcellular; UniProt Q96S37). HPA supplies no ICC-IF images or main location for this record, so this evidence does not validate an IF staining pattern (HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| The known positive kidney section has no proximal tubule rim. | An IHC run or staining condition may have failed; HPA reports proximal tubule microvilli as positive (HPA tissue IHC). | Confirm proximal tubules on the counterstain, then check the antibody instructions, retrieval run, dilution, and detection controls (general IHC practice). Do not infer SLC22A12 specific fixation sensitivity from this result. |
| Brown signal is present but sits mainly in cytoplasm. | Its location conflicts with the expected apical membrane pattern (UniProt Q96S37; HPA tissue IHC). | Examine an intact luminal edge at higher magnification and compare the no primary antibody control (general IHC practice). Record the stain as unconfirmed if a distinct membrane rim cannot be resolved. |
| Negative tissue or an unexpected cell type stains. | The signal may reflect cross-reactivity or endogenous detection activity (general IHC practice); HPA notes disregarded presumed off target binding (HPA tissue IHC). | Compare the reagent omission control and the expected kidney pattern in the same run (general IHC practice). Limit any negative tissue comparison to cells actually listed as not detected by HPA (HPA tissue IHC). |
| The entire section has diffuse chromogen. | Background or detection activity may obscure the distinct membranous pattern (general IHC practice; HPA tissue IHC). | Review blocking, washes, chromogen development, and a no primary antibody control (general IHC practice). Reassess only after individual tubule borders can be seen. |
| A faint rim is difficult to score. | HPA reports a medium level in proximal tubule microvilli, so intensity alone is an incomplete criterion (HPA tissue IHC). | Score the presence and continuity of the luminal rim alongside intensity, using comparable tubules and the same staining run (general IHC practice). |
Experiment evidence
IHC & IF images showing SLC22A12 staining
A02515-2 · IHCIHC analysis of URAT1/SLC22A12 using anti-URAT1/SLC22A12 antibody (A02515-2). URAT1/SLC22A12 was detected in a paraffin-embedded section of
A02515-2 · IHCIHC analysis of URAT1/SLC22A12 using anti-URAT1/SLC22A12 antibody (A02515-2). URAT1/SLC22A12 was detected in a paraffin-embedded section of
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000197891-SLC22A12/tissue/cerebral+cortex#imid_6640248
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000197891-SLC22A12/tissue/colon#imid_6640182
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000197891-SLC22A12/tissue/liver#imid_6640183
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000197891-SLC22A12/tissue/kidney#imid_6640176
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SLC22A12 IHC & IF
🧪Run kidney first: proximal-tubule microvilli should show medium apical staining (HPA: kidney, proximal tubules (microvilli), Medium; UniProt Q96S37: apical membrane). Use adipose tissue, specifically its adipocytes, as the negative tissue (HPA: adipocytes, Not detected); other cells on the kidney section should lack the proximal-tubule apical pattern and serve as internal negatives (UniProt Q96S37: proximal-tubule epithelial expression).
Positive control tissue: Kidney (Proximal tubules (microvilli), 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 SLC22A12; 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 no-primary (secondary-only) and matched rabbit isotype controls, and confirm specificity with SLC22A12 knockout material (selected-SKU IHC caption: rabbit primary and goat anti-rabbit secondary; standard IHC practice). In kidney sections, quench endogenous peroxidase for chromogenic detection and assess tissue autofluorescence for IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A02515-2 paraffin-section caption does not state the fixative (selected-SKU IHC caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0, but that condition alone does not establish retrieval dependence (selected-SKU IHC caption: EDTA pH 8.0). Frozen-section and IF performance cannot be ranked as easier from the supplied evidence; renal autofluorescence may obscure an apical IF pattern (HPA: no ICC-IF cell-line images; UniProt Q96S37: apical membrane; standard IF practice).
HPA tissue IHC evidence for SLC22A12
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium 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 |
|---|---|---|---|---|
| Kidney | Proximal tubules (microvilli) | 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 SLC22A12 IHC Tips
Use renal proximal tubules and their apical staining pattern as the main reference when troubleshooting SLC22A12 chromogenic IHC (HPA tissue IHC; UniProt Q96S37).
What retrieval conditions should I try first for SLC22A12 paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A02515-2). The selected paraffin-section image used that condition before an overnight incubation at 4°C with antibody diluted 1:100 (datasheet A02515-2). Judge retrieval by whether staining resolves along the proximal-tubule apical border, the expected pattern, rather than by total brown signal (HPA tissue IHC; UniProt Q96S37 localisation). If staining is weak, adjust heating time on matched sections while keeping detection and exposure conditions constant; inspect tissue integrity and edge staining after each change (standard IHC practice). Compare the result with a kidney positive control and a primary-antibody omission control (HPA tissue IHC; standard IHC practice).
How should I troubleshoot weak staining when fixation conditions vary?
Target-specific fixation sensitivity for SLC22A12 is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (datasheet A02515-2). Record fixative, duration and processing history for each specimen, then compare matched sections retrieved with EDTA at pH 8.0 under the same conditions (datasheet A02515-2; standard IHC practice). Use the documented 1:100 primary dilution and overnight incubation at 4°C as starting conditions while assessing the effect of fixation (datasheet A02515-2). If one fixation condition yields weak staining, check morphology and the kidney positive control before changing antibody concentration (HPA tissue IHC; standard IHC practice). Do not infer a fixation effect from the protein's membrane topology or tissue distribution (UniProt Q96S37 topology; HPA tissue IHC).
Where should convincing SLC22A12 staining appear in kidney sections?
Expect a distinct membranous signal at the microvilli of renal proximal tubules, consistent with apical cell-membrane localisation (HPA tissue IHC; UniProt Q96S37 localisation). SLC22A12 is detected in proximal-tubule epithelial cells of the renal cortex, and kidney protein expression is reported (UniProt Q96S37 tissue specificity). At 1:100 primary dilution, evaluate whether the chromogenic signal outlines the luminal border rather than filling every cell uniformly (datasheet A02515-2; HPA tissue IHC). Compare neighbouring structures within the same section and review the counterstain to confirm preserved tubular architecture (standard IHC practice). Diffuse cytoplasmic or nuclear colour without an apical pattern should prompt checks for background and tissue damage before assigning target localisation (UniProt Q96S37 localisation; standard IHC practice).
How do isoforms and membrane topology affect epitope interpretation?
SLC22A12 has 4 annotated isoforms and 12 transmembrane segments, so antibody epitope position matters when interpreting staining (UniProt Q96S37 isoforms and topology). The supplied caption identifies the antibody and its 1:100 IHC dilution but does not specify which isoforms or epitope it recognises (datasheet A02515-2). Check the antibody's documented immunogen or epitope before claiming that staining represents every isoform, and assess the result against the expected apical distribution (UniProt Q96S37 isoforms and localisation; standard IHC practice). Glycosylation sites at residues 56 and 102 and phosphothreonine at 542 are annotated, but their effects on this antibody's staining are unreported (UniProt Q96S37 processing). Treat staining differences between specimens as observations until epitope coverage and processing effects are established (standard IHC practice).
How can IF help assess an ambiguous chromogenic pattern?
Use IF as a separate validation experiment; the supplied HPA subcellular record lists no ICC/IF cell images for SLC22A12 (HPA subcellular). Multiplex with a validated proximal-tubule marker and assess whether SLC22A12 follows the expected apical membrane within the same cells (HPA tissue IHC; UniProt Q96S37 localisation; standard IF practice). Select fluorophores after measuring tissue autofluorescence in an unstained section, and include single-label controls to check spectral overlap (standard IF practice). Choose permeabilisation only after establishing whether the antibody epitope faces the extracellular or intracellular side of the membrane; that epitope position is not supplied here (UniProt Q96S37 topology; datasheet A02515-2). Keep any IF interpretation distinct from the paraffin-section chromogenic result and its pH 8.0 retrieval condition (datasheet A02515-2).
What should I check when DAB obscures the apical signal?
The selected paraffin-section workflow used 10% goat-serum blocking, a 1:100 rabbit primary overnight at 4°C, and peroxidase-based DAB detection (datasheet A02515-2). If diffuse brown signal obscures the apical border, compare a primary-antibody omission section and a kidney positive control processed in the same run (standard IHC practice; HPA tissue IHC). Check peroxidase blocking, wash quality and development time as general chromogenic IHC variables, recording any changes to one variable at a time (standard IHC practice). Inspect section edges and damaged areas separately from intact tubules, because concentrated colour there can mislead interpretation (standard IHC practice). HPA reports presumed off-target binding in its tissue assessment, so morphology and location remain essential checks (HPA tissue IHC).
How should I quantify SLC22A12 staining across kidney sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define proximal-tubule epithelial cells and their apical borders as the analysis region before scoring, consistent with the reported expression pattern (HPA tissue IHC; UniProt Q96S37 localisation). Record the percentage of tubules with convincing apical signal and, if intensity is needed, calculate an H-score from prespecified intensity categories (standard IHC practice). Normalise positive counts to the number of evaluable proximal tubules or measured proximal-tubule area, rather than total section area (standard IHC practice). Keep retrieval at EDTA pH 8.0, primary dilution at 1:100, and chromogen development consistent across compared sections (datasheet A02515-2; standard IHC practice). Exclude folds, necrosis and poorly preserved tubules using criteria set before scoring, and report those exclusions (standard IHC practice).
When is brown staining credible evidence of SLC22A12 expression?
The strongest tissue-level pattern is membranous staining at renal proximal-tubule microvilli, matching the reported apical localisation (HPA tissue IHC; UniProt Q96S37 localisation). Treat nuclear colour, uniform staining of unrelated cells, section-edge accentuation and necrotic-area deposits as reasons to investigate artefact before calling a positive (UniProt Q96S37 localisation; standard IHC practice). Check a primary-antibody omission section for endogenous peroxidase or detection-related colour, and compare intact tubules with the kidney positive control (HPA tissue IHC; standard IHC practice). The selected antibody image shows a paraffin section of human skin cancer tissue, while the fixative is unreported (datasheet A02515-2). Interpret staining outside the expected renal pattern cautiously: HPA describes its tissue reliability as Enhanced but also reports presumed off-target binding (HPA tissue IHC).
Boster reagents
Best SLC22A12 / Solute carrier family 22 member 12 IHC Antibodies
A02515-2 has real IHC images from paraffin sections of human skin cancer and mouse and rat kidney (A02515-2 IHC captions). No IF data are supplied (catalog if_image_alts).
Real IHC data
Anti-URAT1/SLC22A12 Antibody
Cat # A02515-2
A02515-2 is listed for IHC and for human, mouse and rat reactivity (catalog applications/reactivity). Its IHC images show paraffin sections of human skin cancer and mouse and rat kidney (A02515-2 IHC captions).
Which to pick: Choose A02515-2 for paraffin-section tissue IHC: it is a rabbit polyclonal antibody with IHC listed and images from those tissues (catalog host/dilution_raw/applications; A02515-2 IHC captions). It is also the cross-species choice here, with human, mouse and rat reactivity listed and IHC images for each species (catalog reactivity; A02515-2 IHC captions). No IF/ICC antibody can be recommended from this payload because A02515-2 has neither a listed IF/ICC application nor an IF image; the fixative for its paraffin sections is unreported (catalog applications/if_image_alts; A02515-2 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96S37 (S22AC_HUMAN, Solute carrier family 22 member 12).
- Human Protein Atlas. SLC22A12 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SLC22A12 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. SLC22A12 antibody validation summary (1 antibodies).
- Low Expression Levels of SLC22A12 Indicates a Poor Prognosis and Progresses Clear Cell Renal Cell Carcinoma. Frontiers in oncology 2021 — PMC8262335.
- 27-Hydroxycholesterol regulates human SLC22A12 gene expression through estrogen receptor action. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2021 — PMC7771643.
- PubMed PMID:12024214 — UniProt-cited evidence.
- PubMed PMID:12975309 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SLC22A12 (Solute carrier family 22 member 12). Expected localisation Proximal-tubule microvillar membrane (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02515-2). Validated IHC antibody: A02515-2 at 1:50 recommended; image 1:100 (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 skin cancer tissue; fixative not specified (datasheet A02515-2); 2. Fixation: Image fixative and duration unreported (datasheet A02515-2); 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 A02515-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02515-2); 8. Primary antibody: Rabbit anti-SLC22A12, 1:50 recommended; image 1:100 (datasheet A02515-2); 9. Primary incubation: Overnight at 4 °C (datasheet A02515-2). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Presumed off-target binding may mimic positive staining (HPA tissue IHC); Isoform / epitope: 4 isoforms; assess epitope position and loop orientation (UniProt); Regulation: No specific regulator reported (UniProt). Evidence: UniProt Q96S37, HPA tissue IHC · Enhanced, HPA ICC-IF · Membrane, Datasheet A02515-2, PMC8262335, PMC7771643. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SLC22A12/immunohistochemistry-ihc-guide.