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SLC34A2 / Sodium-dependent phosphate transport protein 2B · IHC design guide
Design Immunohistochemistry for SLC34A2
Plan paraffin-section IHC for SLC34A2 using lung type II pneumocytes and genital tract ciliated cells as staining references (HPA tissue IHC). The guide covers fixation consistency, staining interpretation and controls for the IHC-validated antibody.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SLC34A2 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 in lung type II pneumocytes and female genital tract (HPA tissue IHC) | |
| Staining pattern | Membranous type II pneumocytes and genital tract ciliated cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03957-1) | |
| Positive control | Bronchus+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 | Ciliary staining may complicate membrane-only scoring (HPA tissue IHC) | |
| Regulation | Lung-enriched RNA expression; no regulator specified (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms; epitope differences unreported; topology matters (UniProt) |
Section 1
Recommended SLC34A2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA retrieval (datasheet: pH 8.0); the four published protocols below provide additional SLC34A2 IHC conditions (PMC8298178; PMC11168907; PMC11083581; PMC9246846).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A03957-1) |
| Fixation | Image fixative and duration unreported (datasheet A03957-1); 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 A03957-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03957-1) |
| Primary antibody | Rabbit anti-SLC34A2, 2-5μg/ml (datasheet A03957-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03957-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03957-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SLC34A2-positive staining in ciliated cells (ciliary rootlets) of bronchus (HPA tissue IHC: High). HPA tissue profile: Membranous expression in lung type II pneumocytes and female genital tract. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet: A03957-1). Published citrate retrieval conditions offer alternatives (PMC8298178; PMC9246846).
Section 2
What Is the Expected SLC34A2 Staining Pattern?
SLC34A2 is an eight-pass membrane transporter expected at the apical surface, including enterocyte brush borders (UniProt O95436 topology and subcellular location). In paraffin-section IHC, look for strong staining in lung alveolar type II cells and membranous staining in the female genital tract; small-intestinal enterocyte microvilli show medium staining (HPA: Enhanced tissue-IHC reliability; HPA: High in alveolar type II cells; HPA: Medium in enterocyte microvilli).
| Strong staining outlines alveolar type II cells or highlights an epithelial apical surface. | This fits the expected IHC pattern when the stained cells and surface can be identified on the section (HPA: High in alveolar type II cells; HPA: membranous tissue profile; UniProt O95436: apical membrane). |
| Nuclear staining dominates a tissue-IHC slide while the expected epithelial membrane is faint. | Treat this as a compartment mismatch and investigate an artefact before scoring it as the expected IHC result (UniProt O95436: apical membrane; HPA: membranous tissue profile). Nuclear signal has separate ICC-IF support and needs its own context (HPA: supported nucleoplasm in ICC-IF). |
| Adipocytes or other cells listed as undetected stain more strongly than the expected epithelial cells. | Consider cross-reactivity or endogenous detection activity, then check the detection controls and tissue morphology (HPA: Not detected in adipocytes of adipose tissue; general IHC practice). Do not infer target expression from color alone. |
| Color spreads across several compartments or obscures cell boundaries. | Diffuse background prevents a confident apical or membranous call (UniProt O95436: apical membrane; HPA: membranous tissue profile). Check blocking, primary-antibody concentration, washing and detection controls as general IHC troubleshooting steps (general IHC practice). |
| A lung section shows no convincing alveolar type II-cell signal. | The expected positive compartment has failed on that slide; review whether alveolar type II cells are present before interpreting other tissue as negative (HPA: High in alveolar type II cells; UniProt O95436: lung expression). Then inspect assay controls and staining conditions (general IHC practice). |
💡Expected SLC34A2 appearanceCall a positive tissue-IHC result when identifiable alveolar type II cells show strong membrane-associated staining, or enterocyte microvilli show medium apical staining; dominant nuclear color or staining in an HPA-undetected cell population calls for investigation (HPA: lung and small-intestine IHC levels; UniProt O95436: apical membrane; HPA: Not detected in adipose-tissue adipocytes).
| Membrane topology and apical position (UniProt O95436 topology and subcellular location) | Eight transmembrane segments support a membrane-centered readout, while the annotated apical location makes epithelial surface orientation useful for interpretation (UniProt O95436 topology and subcellular location). The supplied record does not identify the catalog antibody's epitope, so topology alone cannot predict its retrieval needs. |
| Tissue and cell context (HPA: tissue IHC) | Lung alveolar type II cells are High, enterocyte microvilli are Medium, and endometrial and fallopian-tube ciliated cells are High (HPA: tissue IHC). Match signal to the named cells and structures; a whole-tissue label is less specific than these observations. |
| Airway annotation difference (UniProt O95436; HPA: bronchus IHC) | UniProt reports alveolar type II-cell expression and no bronchiolar epithelial expression, while HPA reports High staining at bronchial ciliated-cell rootlets (UniProt O95436: lung tissue specificity; HPA: High in bronchial ciliated-cell rootlets). Record the sampled structure and morphology rather than merging these distinct observations. |
| Isoforms and glycosylation (UniProt O95436) | The record lists two isoforms and five glycosylation sites at residues 295, 308, 313, 321 and 340 (UniProt O95436: isoforms and glycosylation). Without an antibody epitope or target-specific fixation data, these annotations do not establish an IHC staining difference or retrieval setting. |
| Validation and assay scope (HPA: antibody and subcellular records) | Two rabbit polyclonal antibodies, HPA037989 and HPA066474, have Enhanced IHC status and Approved ICC status (HPA: antibody validation). HPA's ICC-IF record supports a main plasma-membrane location plus nucleoplasm and cytosol; those ICC-IF observations do not replace tissue-IHC cell and compartment criteria (HPA: subcellular ICC-IF; HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive lung lacks a clear signal. | The section may lack identifiable alveolar type II cells, or the IHC run may have underperformed (HPA: High in alveolar type II cells; general IHC practice). | Confirm alveolar morphology and review the run's positive control, retrieval setting, primary-antibody dilution and detection steps (general IHC practice). No target-specific retrieval condition is established by the supplied sources. |
| Membrane edges are weak but diffuse color is strong. | Background can hide the expected apical or membranous pattern (UniProt O95436: apical membrane; HPA: membranous tissue profile; general IHC practice). | Review blocking, washes and primary-antibody dilution; compare a control lacking primary antibody to assess detection background (general IHC practice). |
| Undetected-cell populations stain convincingly. | Cross-reactivity or endogenous detection activity is possible when staining conflicts with the HPA cell-level reference (HPA: Not detected in adipose-tissue adipocytes; general IHC practice). | Verify cell identity and compare appropriate detection controls before calling the signal SLC34A2 (general IHC practice). |
| Nuclear staining appears on tissue IHC. | It does not match the main expected tissue-IHC membrane pattern (HPA: membranous tissue profile; UniProt O95436: apical membrane). | Score the membrane and named cell types separately, and investigate nuclear color with controls (general IHC practice). Do not dismiss the distinct ICC-IF nucleoplasm observation (HPA: supported nucleoplasm in ICC-IF). |
| Bronchial ciliary-rootlet staining seems to conflict with the lung annotation. | HPA reports High bronchial ciliated-cell rootlet staining; UniProt reports absence from bronchiolar epithelium (HPA: bronchus IHC; UniProt O95436: lung tissue specificity). | Document whether the section shows bronchus, bronchiole or alveoli and identify the stained structure before interpreting the discrepancy (HPA: bronchus and lung IHC; UniProt O95436: lung tissue specificity). |
| IF/ICC question: which compartment should be checked? | HPA supports plasma membrane as the main ICC-IF location, with additional nucleoplasm and cytosol (HPA: subcellular ICC-IF). | Interpret those ICC-IF compartments using the separate IF/ICC guide; tissue-IHC scoring here follows the cell-specific membranous pattern (HPA: subcellular ICC-IF; HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing SLC34A2 staining
A03957-1 · IHCIHC analysis of SLC34A2 using anti-SLC34A2 antibody (A03957-1). SLC34A2 was detected in a paraffin-embedded section of human lung cancer tis
A03957-1 · IHCIHC analysis of SLC34A2 using anti-SLC34A2 antibody (A03957-1). SLC34A2 was detected in a paraffin-embedded section of mouse lung tissue. He
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000157765-SLC34A2/tissue/cerebral+cortex#imid_17535651
HPA · LungHuman Protein Atlas IHC, Lung. Source: https://www.proteinatlas.org/ENSG00000157765-SLC34A2/tissue/lung#imid_17535522
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000157765-SLC34A2/tissue/colon#imid_17535577
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000157765-SLC34A2/tissue/liver#imid_17535578
HPA · ICC-IF RPTEC/TERT1Human Protein Atlas immunofluorescence, RPTEC/TERT1, antibody HPA037989; green = SLC34A2, blue = nucleus, red = microtubules.
HPA · ICC-IF RPTEC/TERT1Human Protein Atlas immunofluorescence, RPTEC/TERT1, antibody HPA037989; green = SLC34A2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SLC34A2 IHC & IF
🧪Run lung first: alveolar type II cells should stain (HPA: High in lung alveolar cells type II). Use adipose tissue as the negative, where adipocytes should lack staining (HPA: Not detected in adipocytes); bronchiolar epithelium on the lung slide should also lack target staining (UniProt O95436: lung tissue specificity).
Positive control tissue: Bronchus (Ciliated cells (ciliary rootlets), HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SLC34A2 in RPTEC/TERT1, EFO-21, U2OS, with annotated localisation: Plasma membrane (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a host-species- and clonality-matched isotype control, and SLC34A2 knockout material processed alongside the test section. Quench endogenous peroxidase in lung sections before HRP/DAB detection and check the no-primary slide for residual background (caption: HRP/DAB detection in a paraffin-embedded lung cancer section).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A03957-1 paraffin-section caption does not report a fixative (caption: fixative unreported). The demonstrated IHC procedure used heat retrieval in EDTA at pH 8.0, but a retrieval dependency has not been established (caption: EDTA retrieval). The supplied evidence does not establish whether frozen sections or IF are easier; in lung sections, endogenous peroxidase can complicate chromogenic interpretation (caption: lung tissue and HRP/DAB detection).
HPA tissue IHC evidence for SLC34A2
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 |
|---|---|---|---|---|
| Bronchus | Ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
| Endometrium | Ciliated epithelial cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (cilia axoneme) | High | Protein (IHC) | HPA → |
| Lung | Alveolar cells type II | High | Protein (IHC) | HPA → |
| Small intestine | Enterocytes - 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 | Endocrine 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 SLC34A2 IHC Tips
Troubleshoot SLC34A2 staining in paraffin sections by checking retrieval, membrane localisation, cell identity and detection controls before comparing staining intensity.
What retrieval should I try first when SLC34A2 staining is weak?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A03957-1). The selected tissue-IHC image used this retrieval with 2 μg/ml primary antibody overnight at 4°C, so use those conditions as the initial comparison point (datasheet A03957-1). If staining remains weak, compare retrieval durations on adjacent sections while keeping antibody concentration and detection conditions constant; the caption gives no retrieval duration (datasheet A03957-1). Judge improvement by staining in the expected cell population and membrane compartment, rather than by stronger staining throughout the section (HPA: high in lung alveolar type II cells; UniProt O95436: apical cell membrane).
Could fixation explain weak or uneven SLC34A2 staining?
The selected paraffin-section caption does not state a fixative, so target-specific fixation sensitivity is unknown (datasheet A03957-1). Record the actual fixative and processing history for each specimen, then compare sections processed together before assigning a difference to SLC34A2 expression (standard IHC practice). Examine folds, detached tissue and poorly preserved areas before scoring; these can produce uneven chromogenic staining independent of the target (standard IHC practice). Keep retrieval at EDTA pH 8.0 and the primary at 2 μg/ml when comparing specimens, since both conditions are documented for this antibody (datasheet A03957-1).
Where should convincing SLC34A2 staining appear in tissue?
Prioritise cell-surface staining, especially an apical pattern where tissue polarity is visible: SLC34A2 is assigned to the apical membrane and enterocyte brush border (UniProt O95436 localisation). Lung alveolar type II cells show high staining, and small-intestinal enterocyte microvilli show medium staining in the tissue profile (HPA: lung and small intestine). The profile also reports staining at ciliary structures in bronchus and fallopian tube, so interpret those sites by their recorded cell and subcellular patterns (HPA: ciliary rootlets and cilia axoneme). Diffuse staining alone deserves control review before it is counted as the expected tissue-IHC pattern (UniProt O95436 localisation; standard IHC practice).
How do epitope location and isoforms affect a negative result?
SLC34A2 has 2 annotated isoforms, but the supplied antibody caption does not map its epitope or establish which isoforms it detects (UniProt O95436 isoforms; datasheet A03957-1). Its 8 transmembrane segments separate extracellular and cytoplasmic regions, so epitope accessibility may depend on processing and retrieval (UniProt O95436 topology; standard IHC practice). Five annotated glycosylation sites, at residues 295, 308, 313, 321 and 340, lie within the annotated extracellular region spanning residues 234–362 (UniProt O95436 topology and glycosylation). A negative section therefore cannot establish isoform absence or a glycosylation effect without antibody epitope information and suitable tissue controls (standard IHC interpretation).
How should I assess SLC34A2 in a multiplex IF experiment?
For a lung experiment, pair SLC34A2 with an independently validated alveolar type II cell marker and check whether membrane signal belongs to those cells (HPA: high in alveolar type II cells; UniProt O95436: apical membrane). Choose fluorophores after inspecting unstained tissue autofluorescence in every intended channel, and include single-label controls to assess spectral spillover (standard IF practice). Permeabilisation should follow a mapped epitope: an extracellular epitope may be accessible without it, whereas a cytoplasmic epitope requires membrane access (UniProt O95436 topology; standard IF practice). The supplied IHC caption does not establish this antibody’s IF performance or epitope location (datasheet A03957-1).
How can I reduce diffuse brown staining without losing the membrane signal?
First inspect a section without primary antibody to separate detection-system background from antibody-dependent staining (standard IHC practice). The selected image used 10% goat serum blocking, 2 μg/ml primary overnight at 4°C, and a peroxidase-conjugated secondary for 30 minutes at 37°C (datasheet A03957-1). Apply an endogenous peroxidase block and assess the DAB development time as general chromogenic IHC controls, especially when the no-primary section shows colour (standard IHC practice). If background persists only with primary antibody, titrate it on matched sections and retain the condition that preserves expected cell-surface staining (UniProt O95436 localisation; standard IHC practice).
What should I score when comparing SLC34A2 across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population before measuring colour; lung alveolar type II cells are a documented positive population (HPA: high in alveolar type II cells). For each region, record the percentage of eligible cells with convincing membrane staining and, if intensity matters, calculate an H-score from percentages at intensity grades 0–3, giving a range of 0–300 (standard IHC scoring practice). Normalise positive counts to the number of eligible viable cells, or report stained membrane area per mm² of eligible viable tissue (standard IHC quantification practice). Keep retrieval, imaging and DAB development consistent across compared sections (datasheet A03957-1 retrieval; standard IHC practice).
How do I distinguish true SLC34A2 staining from artefact?
A credible result combines the expected cell population with cell-surface staining: lung alveolar type II cells are high in the tissue profile, and the protein is assigned to the apical membrane (HPA: lung; UniProt O95436 localisation). Review diffuse nuclear colour cautiously in chromogenic tissue sections; supported nucleoplasmic and cytosolic locations come from subcellular IF data and do not by themselves validate a tissue-IHC pattern (HPA subcellular). Exclude tissue edges and necrotic areas from interpretation when staining follows damage rather than intact cells (standard IHC practice). Check a no-primary section for endogenous enzyme or detection background before calling unexpected staining positive (standard IHC practice).
Boster reagents
Best SLC34A2 / Sodium-dependent phosphate transport protein 2B IHC Antibodies
A03957-1 has IHC images from paraffin sections of human lung cancer, mouse lung and rat lung (A03957-1 IHC image captions).
Real IHC data
Anti-SLC34A2 Antibody ®
Cat # A03957-1
A03957-1 is listed for IHC in human, mouse and rat (catalog: applications and reactivity). Its IHC images show paraffin sections of human lung cancer, mouse lung and rat lung (A03957-1 IHC image captions).
Which to pick: Choose A03957-1 for chromogenic IHC on paraffin sections; its images document EDTA retrieval at pH 8.0, 2 μg/ml primary antibody and peroxidase/DAB detection (A03957-1 IHC image captions). It is the available cross-species choice for human, mouse and rat tissue (catalog: reactivity; A03957-1 IHC image captions). No IF/ICC-validated SKU is supplied (catalog: applications and IF image captions); fixation and clonality are unreported (A03957-1 IHC image captions; catalog: clone).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O95436 (NPT2B_HUMAN, Sodium-dependent phosphate transport protein 2B).
- Human Protein Atlas. SLC34A2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SLC34A2 subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the nucleoplasm and cytosol..
- Human Protein Atlas. SLC34A2 antibody validation summary (2 antibodies).
- Evaluation of the Prognostic Value of Solute Carrier Family 34 Member 2 "SLC34A2" in Papillary Thyroid Carcinoma: An Immunohistochemical Study. Analytical cellular pathology (Amsterdam) 2021 — PMC8298178.
- SLC34A2 promotes cell proliferation by activating STX17-mediated autophagy in esophageal squamous cell carcinoma. Thoracic cancer 2024 — PMC11168907.
- SLC34A2 Targets in Calcium/Phosphorus Homeostasis of Mammary Gland and Involvement in Development of Clinical Mastitis in Dairy Cows. Animals : an open access journal from MDPI 2024 — PMC11083581.
- Phosphate dysregulation via the XPR1-KIDINS220 protein complex is a therapeutic vulnerability in ovarian cancer. Nature cancer 2022 — PMC9246846.
- PubMed PMID:10329428 — UniProt-cited evidence.
- PubMed PMID:10610722 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SLC34A2 (Sodium-dependent phosphate transport protein 2B). Expected localisation Membranous in lung type II pneumocytes and female genital tract (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03957-1). Validated IHC antibody: A03957-1 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Ciliated cells (ciliary rootlets)); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A03957-1); 2. Fixation: Image fixative and duration unreported (datasheet A03957-1); 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 A03957-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03957-1); 8. Primary antibody: Rabbit anti-SLC34A2, 2-5μg/ml (datasheet A03957-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03957-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Ciliary staining may complicate membrane-only scoring (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences unreported; topology matters (UniProt); Regulation: Lung-enriched RNA expression; no regulator specified (HPA tissue RNA). Evidence: UniProt O95436, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the plasma membrane. In addition localized to…, Datasheet A03957-1, PMC8298178, PMC11168907. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SLC34A2/immunohistochemistry-ihc-guide.