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SLC11A2 / Natural resistance-associated macrophage protein 2 · IHC design guide
Design Immunohistochemistry for SLC11A2
This guide plans chromogenic paraffin IHC with antibody A02622 at 1:50–1:200 (datasheet A02622). Use salivary gland glandular cells as a high-staining reference and adipocytes as a negative comparison while assessing cytoplasmic and membranous signal (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SLC11A2 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 | Variable cytoplasmic and membranous tissue staining (HPA tissue IHC) | |
| Staining pattern | Glandular and other cells: variable cytoplasmic/membranous stain (HPA tissue IHC) | |
| Antigen retrieval | Tris pH 9.0 HIER, heat-mediated (datasheet A02622) | |
| Positive control | Salivary gland+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 formalin fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining has low concordance with RNA expression (HPA tissue IHC) | |
| Regulation | Iron/cobalt-linked neuronal degradation (UniProt) | |
| Isoform / epitope | Five isoforms; verify epitope conservation and membrane side (UniProt) |
Section 1
Recommended SLC11A2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is complemented by published SLC11A2 tissue-staining methods with usable parameters (PMC13576891; PMC12845770).
| Sample | Paraffin-embedded rat stomach tissue; fixative not specified (datasheet A02622) |
| Fixation | Image fixative and duration unreported (datasheet A02622); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris pH 9.0 (datasheet A02622); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-SLC11A2, 1:50-1:200 (datasheet A02622) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SLC11A2-positive staining in glandular cells of salivary gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and membranous expression at variable levels in several different cell types. No signal in the no-primary control. |
💡Decision noteStart with heat retrieval in Tris pH 9.0 (datasheet A02622); the automated published method used CC1 for 32 min (PMC13576891).
Section 2
What Is the Expected SLC11A2 Staining Pattern?
SLC11A2 is a 12-pass membrane protein found predominantly in early endosomes beneath the apical membrane of polarized epithelia, with other membrane locations annotated (UniProt P49281 topology and subcellular location). In paraffin sections, expect variable cytoplasmic and membranous staining; salivary glandular cells provide the strongest listed example (HPA tissue IHC: High). Interpret any pattern cautiously because tissue IHC reliability is Uncertain (HPA tissue IHC).
| Salivary glandular cells show distinct cytoplasmic and membranous chromogenic staining. | This fits the strongest listed tissue example, rated High, and HPA’s overall cytoplasmic and membranous profile (HPA tissue IHC). An apical or punctate intracellular emphasis is biologically plausible from early-endosome localization beneath the apical membrane; routine chromogenic IHC alone cannot identify individual organelles (UniProt P49281 subcellular location; standard IHC practice). |
| Glandular cells or respiratory epithelial cells stain less strongly than the salivary example. | Medium staining is reported in adrenal, breast, cervix and colon glandular cells and bronchial respiratory epithelial cells; duodenal and small-intestinal glandular cells are listed as Low (HPA tissue IHC). Compare the actual cell population with its corresponding HPA entry. Variable intensity across tissues is consistent with the HPA profile, but staining strength alone cannot establish specificity (HPA tissue IHC: Uncertain). |
| Signal is concentrated in nuclei or appears as a uniform nuclear stain. | A predominantly nuclear pattern conflicts with the annotated membrane locations and 12 transmembrane segments (UniProt P49281 topology and subcellular location). Treat it as suspect: inspect morphology and staining controls, then reassess antibody specificity and detection background (standard IHC practice). Some intracellular signal is expected, but a nuclear-only result does not match the supplied localization evidence (UniProt P49281; HPA tissue IHC). |
| Strong staining appears in a cell population HPA lists as Not detected. | Examples include adipocytes, lymph-node germinal-center cells and smooth-muscle cells (HPA tissue IHC: Not detected). Check cell identity and compare a no-primary control: cross-reactivity or endogenous detection activity can mimic target staining (standard IHC practice). A discrepant field is a reason to investigate, not proof of absence or artifact, because HPA rates tissue IHC reliability Uncertain (HPA tissue IHC). |
| The section shows diffuse haze, or salivary glandular cells show no discernible signal. | Diffuse staining obscures the cytoplasmic and membranous pattern reported by HPA; examine no-primary background and staining distribution (HPA tissue IHC; standard IHC practice). Absent signal in the listed High salivary glandular population prompts a review of section quality, retrieval and detection controls (HPA tissue IHC: High; standard IHC practice). One negative section cannot establish target absence (standard IHC practice). |
💡Expected SLC11A2 appearanceCall a result provisionally positive when salivary glandular cells show distinct High cytoplasmic and membranous staining, potentially with apical or punctate emphasis; uniform nuclear signal or diffuse haze is suspect (HPA tissue IHC: High, Uncertain; UniProt P49281 subcellular location; standard IHC practice).
| Cell population and tissue context | HPA reports variable cytoplasmic and membranous staining, with High salivary glandular staining, several Medium populations and several Not detected populations (HPA tissue IHC). Score named cell populations rather than treating an entire tissue as uniformly positive or negative; HPA’s tissue IHC reliability is Uncertain (HPA tissue IHC; standard IHC practice). |
| Membrane topology and antibody epitope | SLC11A2 has 12 transmembrane segments and both cytoplasmic and extracellular regions (UniProt P49281 topology). The supplied record does not identify the antibody epitope, so topology cannot predict which antigen-retrieval condition will work. Compare retrieval conditions using appropriate controls as a general IHC optimization step, without assuming SLC11A2-specific fixation sensitivity (standard IHC practice). |
| Isoform coverage | UniProt lists 5 isoforms (UniProt P49281 isoforms). The supplied evidence does not map the catalog antibody’s epitope to those isoforms, so a stained section cannot establish which isoform contributed the signal. Report staining as SLC11A2-associated unless separate antibody characterization supports a narrower interpretation (UniProt P49281 isoforms; standard IHC practice). |
| IF/ICC Q&A: should the IHC pattern match fluorescence images? | HPA ICC-IF reports a supported mitochondrial localization and lists A-431, U-251MG and U2OS images (HPA subcellular ICC-IF). UniProt also annotates the mitochondrial outer membrane, alongside endosomal and other membranes (UniProt P49281 subcellular location). These observations can inform interpretation, but fluorescence localization does not establish an IHC-P protocol or validate every chromogenic tissue signal (HPA subcellular ICC-IF; standard IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in salivary glandular cells. | A technical failure is possible even though this population is listed as High; HPA tissue IHC reliability remains Uncertain (HPA tissue IHC; standard IHC practice). | Check section integrity and the run’s positive and detection controls; then review retrieval and antibody conditions using the catalog IHC-P instructions (standard IHC practice). |
| Only nuclear staining is visible. | Nuclear-only staining disagrees with the supplied membrane localization and topology (UniProt P49281 topology and subcellular location). | Inspect morphology and no-primary background, then compare with an independently validated antibody if available; do not score nuclear signal as the expected pattern (standard IHC practice; UniProt P49281). |
| Diffuse brown background hides cell boundaries. | Nonspecific reagent binding or detection background may obscure the variable cytoplasmic and membranous pattern (standard IHC practice; HPA tissue IHC). | Review no-primary control and wash and blocking conditions; score only signal that remains distinguishable in the relevant cells (standard IHC practice). |
| Strong signal appears in HPA Not detected cells. | Cell misidentification, antibody cross-reactivity or endogenous detection activity may explain the discrepancy; an HPA Not detected entry is not an absolute exclusion (HPA tissue IHC: Uncertain; standard IHC practice). | Confirm the cell population and check a no-primary control. For chromogenic detection, assess endogenous enzyme background with an appropriate control (standard IHC practice). |
| Duodenal or small-intestinal glandular staining is faint. | Both populations are listed as Low, so a weak result can agree with the tissue profile (HPA tissue IHC: Low). | Compare them with a stronger listed population and the same run’s controls before changing retrieval or detection conditions (HPA tissue IHC: High in salivary glandular cells; standard IHC practice). |
| Punctate signal and apparent surface staining differ between cells. | SLC11A2 has early-endosome and apical membrane annotations, while HPA reports variable cytoplasmic and membranous tissue staining (UniProt P49281 subcellular location; HPA tissue IHC). | Record compartment, cell type and intensity separately. Seek additional localization evidence before assigning individual chromogenic puncta to a named organelle (standard IHC practice). |
Experiment evidence
IHC & IF images showing SLC11A2 staining
A02622 · IHCImmunohistochemistry of paraffin-embedded rat stomach using DMT1/SLC11A2 Rabbit pAb at dilution of 1:100 .Perform microwave antigen retrieva
A02622 · IHCImmunohistochemistry of paraffin-embedded mouse stomach using DMT1/SLC11A2 Rabbit pAb at dilution of 1:100 .Perform microwave antigen retrie
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000110911-SLC11A2/tissue/cerebral+cortex#imid_10798783
HPA · Salivary glandHuman Protein Atlas IHC, Salivary gland. Source: https://www.proteinatlas.org/ENSG00000110911-SLC11A2/tissue/salivary+gland#imid_10798731
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000110911-SLC11A2/tissue/colon#imid_10798709
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000110911-SLC11A2/tissue/liver#imid_10798718
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA032140; green = SLC11A2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA032140; green = SLC11A2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SLC11A2 IHC & IF
🧪Run salivary gland first and score its glandular cells, where SLC11A2 staining is High (HPA: salivary gland, glandular cells, High). Use adipose tissue adipocytes as the negative comparator (HPA: adipose tissue, adipocytes, Not detected); cells without specific staining on the salivary gland slide should show counterstain and background only, without assuming every adjacent cell type is negative.
Positive control tissue: Salivary gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SLC11A2 in A-431, U-251MG, U2OS, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit polyclonal IgG isotype control (selected-SKU caption: Rabbit pAb), then confirm target specificity with SLC11A2 knockout tissue or a validated immunizing-peptide block. For chromogenic IHC, quench endogenous peroxidase and assess background in salivary gland lumina (HPA: salivary gland, glandular cells, High).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence; the selected-SKU paraffin-section caption does not state a fixative (selected-SKU caption: fixative not stated). That caption uses microwave retrieval in 10 mM Tris/Tris, pH 9.0, for rat stomach at 1:100, so it provides a starting condition rather than evidence that retrieval is required for salivary gland (selected-SKU caption: rat stomach IHC). The supplied evidence does not establish whether frozen sections or IF are easier; assess salivary gland luminal deposits separately from glandular-cell staining (HPA: salivary gland, glandular cells, High).
HPA tissue IHC evidence for SLC11A2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low 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 |
|---|---|---|---|---|
| Salivary gland | Glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SLC11A2 IHC Tips
Troubleshoot SLC11A2 staining in paraffin sections by checking retrieval, compartment pattern and tissue controls before comparing chromogenic signal between samples.
What retrieval should I try when SLC11A2 staining is weak in paraffin sections?
Start with heat-mediated Tris pH 9.0 antigen retrieval for paraffin sections (datasheet A02622). The selected image used microwave retrieval in 10 mM Tris/Tris, pH 9.0, with antibody at 1:100 on paraffin-embedded rat stomach (A02622 tissue-IHC caption). SLC11A2 has 12 transmembrane segments and loops on both sides of the membrane, so assess signal and morphology together when adjusting heat exposure (UniProt P49281 topology; standard IHC practice). If staining remains weak, test another retrieval buffer as a documented fallback, changing one condition at a time and comparing sections processed together (standard IHC practice).
How should I troubleshoot fixation when SLC11A2 staining varies between paraffin blocks?
Target-specific fixation sensitivity for SLC11A2 is unknown from the supplied evidence; the selected paraffin-section image does not report a fixative (A02622 tissue-IHC caption). Record the fixative, fixation duration, tissue thickness and processing history for each block before attributing variable staining to the antibody (standard IHC practice). Compare adjacent sections with the same Tris pH 9.0 retrieval and antibody conditions, using morphology and a concurrently stained control to assess processing effects (datasheet A02622; standard IHC practice). Avoid inferring fixation tolerance from tissue staining patterns or membrane topology; neither establishes how this epitope responds to fixation (HPA tissue IHC; UniProt P49281 topology).
Should SLC11A2 appear at the apical surface or as cytoplasmic puncta?
Both patterns can be plausible in chromogenic sections: SLC11A2 occurs at the apical membrane and predominantly in early endosomes beneath it in polarized epithelia (UniProt P49281 localisation). Endosomal staining can appear punctate within the cytoplasm, while HPA describes variable cytoplasmic and membranous tissue staining (UniProt P49281 localisation; HPA tissue IHC). Assess the stained cell type and its orientation before calling a diffuse edge deposit apical; compare serial sections and counterstained morphology (standard IHC practice). HPA also reports supported mitochondrial localisation by ICC/IF, but its tissue IHC reliability is Uncertain, so compartment assignment in sections needs independent controls (HPA subcellular; HPA tissue IHC).
Could an isoform or membrane sidedness explain a missing SLC11A2 signal?
SLC11A2 has 5 annotated isoforms and 12 transmembrane segments, so identify the catalog antibody's immunogen before assuming it detects every form (UniProt P49281 isoforms; UniProt P49281 topology; standard IHC practice). The supplied evidence gives no A02622 epitope sequence or isoform coverage, leaving those properties unresolved for this antibody (A02622 tissue-IHC caption). Extracellular residues 315–376 include glycosylation sites 336 and 349, whereas the terminal 526–568 region is cytoplasmic (UniProt P49281 topology; UniProt P49281 glycosylation). Compare sections processed together and document retrieval and staining conditions before assigning an absent or shifted pattern to isoform expression (standard IHC practice).
How can IF help check a puzzling chromogenic SLC11A2 pattern?
Use IF/ICC as a complementary localisation check, while evaluating the paraffin-section result against its own controls (standard IF/IHC practice). Multiplex SLC11A2 with a marker of the expected cell type, such as a glandular-cell marker in salivary gland, and inspect single channels before interpreting overlap (HPA tissue IHC; standard IF practice). Choose fluorophores in channels with low tissue autofluorescence and include an unstained section to distinguish fluorescence from specimen background (standard IF practice). Determine whether the antibody epitope faces cytoplasm or the lumen before selecting permeabilisation, because SLC11A2 loops face both sides of its 12-span membrane (UniProt P49281 topology; standard IF practice).
What should I check if chromogenic SLC11A2 staining is diffuse or widespread?
Run a no-primary control through peroxidase detection and chromogen development to locate background generated by the section or detection system (standard IHC practice). Check blocking, washes and antibody concentration, using the caption's 1:100 as the documented starting dilution for A02622 rather than assuming it suits every specimen (A02622 tissue-IHC caption; standard IHC practice). Review endogenous peroxidase blocking and inspect edges, folds and damaged regions separately from intact tissue (standard chromogenic IHC practice). Genuine SLC11A2 staining can be cytoplasmic or membranous, but HPA rates its tissue IHC reliability Uncertain; corroborate widespread signal with tissue and reagent controls (HPA tissue IHC; standard IHC practice).
How should I score SLC11A2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring because HPA reports variable cytoplasmic and membranous staining across cell types (HPA tissue IHC). For intensity, report an H-score from the percentage of cells at each staining level; for prevalence, report percent positive cells or positive-cell density per mm² (standard IHC practice). Normalize counts to the eligible cells or intact tissue area, and keep section selection, threshold and counterstain rules consistent across samples (standard IHC practice). Report membrane and cytoplasmic scores separately when both are visible, and state the controls used because HPA tissue IHC reliability is Uncertain (HPA tissue IHC; standard IHC practice).
How can I distinguish convincing SLC11A2 signal from staining artefact?
Look for staining in plausible cells and compartments: HPA reports high signal in salivary gland glandular cells, while UniProt places SLC11A2 at membranes and in endosomes (HPA tissue IHC; UniProt P49281 localisation). Treat an isolated nuclear pattern, section-edge rim or signal concentrated in necrotic tissue as suspect, then inspect morphology and a no-primary control (UniProt P49281 localisation; standard IHC practice). Exclude endogenous enzyme signal with a detection control before attributing brown deposits to SLC11A2 (standard chromogenic IHC practice). Tissue expression alone is insufficient confirmation because HPA rates its SLC11A2 tissue IHC reliability Uncertain owing to low staining–RNA consistency (HPA tissue IHC).
Boster reagents
Best SLC11A2 / Natural resistance-associated macrophage protein 2 IHC Antibodies
The catalog lists human-, mouse-, and rat-reactive SLC11A2 antibodies for IHC, with paraffin-section images from rat and mouse stomach; IF is listed for one antibody without an image (catalog applications; A02622 image captions).
Real IHC data
Anti-SLC11A2 Antibody
Cat # A02622
A02622 is listed for IHC in human, mouse, and rat, with paraffin-section images from rat and mouse stomach (catalog applications and reactivity; A02622 image captions). A02622-2 is listed for IHC and IF in all three species, but has no supplied IHC or IF image (catalog applications, reactivity, and image alts).
Which to pick: For paraffin-section IHC, choose A02622: its own captions document staining in rat and mouse stomach; the fixative is unreported (A02622 image captions). For IF, choose A02622-2 because IF is listed at 1:50; ICC is unlisted and no IF image is supplied (A02622-2 catalog applications, dilution, and image alts). Both are rabbit polyclonal antibodies listed as reactive with human, mouse, and rat, so species coverage alone does not distinguish them (catalog host, dilution_raw, and reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P49281 (NRAM2_HUMAN, Natural resistance-associated macrophage protein 2).
- Human Protein Atlas. SLC11A2 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. SLC11A2 subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. SLC11A2 antibody validation summary (2 antibodies).
- SLC11A2: a promising biomarker and therapeutic target in ovarian cancer. Scientific reports 2023 — PMC9860018.
- Functional validation of driver mutation-specific uveal melanoma biomarkers: role of COL9A3 in cancer cell plasticity. The Journal of pathology 2026 — PMC13576891.
- Dietary Glyphosate Exposure Disrupts Hepatic and Reproductive Function in Female Zebrafish at Regulatory Safe Levels. Toxics 2026 — PMC12845770.
- MAFF regulates ferroptotic sensitivity through iron homeostasis and fatty acid synthesis. Cell death & disease 2026 — PMC13402699.
- PubMed PMID:9464519 — UniProt-cited evidence.
- PubMed PMID:9790986 — UniProt-cited evidence.
- PubMed PMID:9642100 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SLC11A2 (Natural resistance-associated macrophage protein 2). Expected localisation Variable cytoplasmic and membranous tissue staining (HPA tissue IHC). Antigen retrieval: Tris pH 9.0 HIER, heat-mediated (datasheet A02622). Validated IHC antibody: A02622 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Salivary gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat stomach tissue; fixative not specified (datasheet A02622); 2. Fixation: Image fixative and duration unreported (datasheet A02622); 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: Tris pH 9.0 (datasheet A02622); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-SLC11A2, 1:50-1:200 (datasheet A02622); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep formalin fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining has low concordance with RNA expression (HPA tissue IHC); Isoform / epitope: Five isoforms; verify epitope conservation and membrane side (UniProt); Regulation: Iron/cobalt-linked neuronal degradation (UniProt). Evidence: UniProt P49281, HPA tissue IHC · Uncertain, HPA ICC-IF · mitochondria., Datasheet A02622, PMC13576891, PMC12845770. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SLC11A2/immunohistochemistry-ihc-guide.