SLC11A1 / Natural resistance-associated macrophage protein 1 · IHC design guide

Design Immunohistochemistry for SLC11A1

Plan SLC11A1 staining in paraffin sections using the catalog antibody’s IHC conditions (datasheet A02547-3). Expect variable cytoplasmic staining, including in subsets of immune cells, and interpret cell-level results cautiously because antibody staining has low concordance with RNA expression (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for SLC11A1 (IHC for SLC11A1): expected localisation Variable cytoplasmic staining (HPA tissue IHC)., antibody A02547-3, validated IHC image, and IHC protocol steps — Printable **SLC11A1** IHC protocol sheet — expected localisation Variable cytoplasmic staining (HPA tissue IHC)., antibody A02547-3, controls and protocol steps. Open the full SLC11A1 IHC guide →

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SLC11A1 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 staining (HPA tissue IHC).
	Staining pattern	Cytoplasmic staining in subsets of immune cells (HPA tissue IHC).
	Antigen retrieval	Citrate pH 6 HIER, heat-mediated (datasheet A02547-3)
	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 (standard IHC practice; not target-specific); target-specific effects are unknown.
	Caveat	Low antibody–RNA concordance complicates interpretation (HPA tissue IHC).
	Regulation	Stimulus-dependent staining intensity is unreported (UniProt).
	Isoform / epitope	2 isoforms; epitope coverage is unknown; loop position matters (UniProt).

Section 1

Recommended SLC11A1 IHC & IF Protocols

The catalog antibody’s IHC-P protocol is accompanied by published SLC11A1 staining methods for mouse hippocampus, glioma samples, and rat choroid plexus (PMC12808816; PMC9748290; PMC4412010).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human intestinal cancer tissues; fixative not specified (datasheet A02547-3)
Fixation	Image fixative and duration unreported (datasheet A02547-3); 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 A02547-3)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A02547-3)
Primary antibody	Rabbit anti-SLC11A1, 0.5-1μg/ml (datasheet A02547-3)
Primary incubation	Overnight at 4 °C (datasheet A02547-3)
Detection	Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02547-3)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	SLC11A1-positive staining in respiratory epithelial cells of bronchus (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression at variable levels in several tissues, including subsets of immune cells in most tissues. No signal in the no-primary control.

💡Decision noteStart with heat-mediated citrate pH 6 retrieval for the catalog antibody (datasheet A02547-3); the mouse hippocampus protocol also reports boiling in citrate pH 6.0 (PMC12808816).

Section 2

What Is the Expected SLC11A1 Staining Pattern?

SLC11A1 is a 12-pass membrane protein of phagosomes, late endosomes and lysosomes (UniProt P49279 topology and subcellular location). Expect cytoplasmic staining in macrophages and subsets of immune cells, with variable staining in other cells (UniProt P49279 tissue specificity; HPA tissue IHC). Interpret apparent positives cautiously: HPA rates its tissue IHC profile Uncertain because antibody staining has low consistency with RNA expression (HPA tissue IHC).

What am I looking at on my slide?
Cytoplasmic, granular staining in scattered lung macrophages.	Consistent with vesicular localisation (UniProt P49279 subcellular location). HPA reports low staining in lung macrophages, so a faint subset of cells can be plausible (HPA tissue IHC). Assess cell identity and morphology before scoring.
Predominantly nuclear or continuous cell-surface staining in a proposed positive cell.	That pattern does not match the phagosome, late-endosome and lysosome membrane locations reported by UniProt (UniProt P49279 subcellular location). Check the pattern against controls; compartment mismatch alone does not identify the source of staining.
Broad staining of adipocytes or cardiomyocytes.	HPA reports these cells as not detected (HPA tissue IHC). Consider cross-reactivity or endogenous chromogenic activity (standard IHC practice), while allowing for HPA’s Uncertain tissue profile (HPA tissue IHC).
Uniform haze across tissue and spaces between cells.	Diffuse background makes cell-specific cytoplasmic staining hard to judge. Review blocking, washes and detection controls for nonspecific signal (standard IHC practice); this appearance cannot establish SLC11A1 localisation.
No staining in bronchial respiratory epithelial cells.	HPA reports medium staining in this cell population (HPA tissue IHC). A blank section calls for a technical review, but the HPA profile is Uncertain; one negative field does not prove assay failure (HPA tissue IHC).

💡Expected SLC11A1 appearanceA plausible IHC positive is cell-associated, variable cytoplasmic staining, including granular signal in lung macrophages; HPA calls that macrophage signal low, while uniform haze or widespread staining of HPA-negative adipocytes is suspect (UniProt P49279 subcellular location; HPA tissue IHC; standard IHC practice).

How each factor affects the staining
Tissue and cell selection	UniProt lists macrophages and peripheral blood leukocytes, plus lung, spleen and liver (UniProt P49279 tissue specificity). HPA reports diverse stained cell types and an Uncertain IHC profile; identify the scored cell population rather than treating a whole tissue as uniformly positive (HPA tissue IHC).
Compartment and antibody epitope	The protein has 12 transmembrane segments and cytoplasmic and extracellular regions (UniProt P49279 topology). The supplied record does not locate this antibody’s epitope; topology alone cannot predict its retrieval requirement or staining strength.
Protein forms	UniProt lists 2 isoforms, one chain spanning residues 1–550, and glycosylation sites at 324 and 338 (UniProt P49279). Without an antibody epitope or isoform-specific validation, do not assign different IHC intensities to these features.
IHC evidence strength	HPA rates the tissue profile Uncertain; its antibody HPA029590 is also rated Uncertain for IHC (HPA tissue IHC; HPA antibodies). Medium staining in a listed cell type is an observation, not independent confirmation of specificity.
IF/ICC: Should mitochondrial or nucleolar signal be expected?	HPA reports approved mitochondrial and additional nucleolar ICC-IF localisation, whereas UniProt places SLC11A1 on phagosomal, late-endosomal and lysosomal membranes (HPA subcellular ICC-IF; UniProt P49279 subcellular location). Treat this as a source discrepancy; an IF/ICC result does not settle an IHC localisation call.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Weak or absent signal in bronchial respiratory epithelium.	HPA reports medium staining there, but its overall IHC profile is Uncertain (HPA tissue IHC); the result may also reflect an IHC workflow problem (standard IHC practice).	Check tissue morphology, assay controls, antibody use, retrieval and detection against the validated IHC-P instructions (standard IHC practice). Do not infer a SLC11A1-specific fixation effect from this result.
Lung macrophages show only faint or scattered staining.	HPA reports low staining in lung macrophages (HPA tissue IHC).	Score identifiable macrophages and compare like cell populations across sections; avoid calling the whole lung negative solely because other cells lack signal (HPA tissue IHC; standard IHC practice).
Adipocytes or cardiomyocytes stain broadly.	Both cell populations are listed as not detected by HPA (HPA tissue IHC); nonspecific antibody or detection signal is possible (standard IHC practice).	Inspect a matched negative detection control and tissue morphology, then reassess specificity. HPA’s Uncertain IHC rating limits how decisively its negatives can be used (HPA tissue IHC).
A brown deposit obscures cell boundaries.	Diffuse chromogen can reflect background or endogenous detection activity (standard IHC practice).	Review blocking, washes and the detection control; interpret only signal assignable to intact cells (standard IHC practice). Avoid scoring deposits as cytoplasmic SLC11A1.
Signal is mainly nuclear or outlines the plasma membrane.	Those appearances conflict with UniProt’s phagosome, late-endosome and lysosome membrane annotation (UniProt P49279 subcellular location).	Compare the suspicious region with controls and examine cell morphology before calling it positive (standard IHC practice); record the compartment mismatch with the result.
IF/ICC and tissue IHC suggest different compartments.	HPA’s approved ICC-IF locations are mitochondria and nucleoli, while UniProt lists phagosomal, late-endosomal and lysosomal membranes (HPA subcellular ICC-IF; UniProt P49279).	Report the antibody, application and observed compartment separately; keep the IHC interpretation qualified by HPA’s Uncertain tissue profile (HPA tissue IHC).

Source: UniProt P49279 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing SLC11A1 staining

IHC analysis of SLC11A1 using anti-SLC11A1 antibody (A02547-3). SLC11A1 was detected in paraffin-embedded section of human intestinal cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-SLC11A1 Antibody (A02547-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

A02547-3 · IHCIHC analysis of SLC11A1 using anti-SLC11A1 antibody (A02547-3). SLC11A1 was detected in paraffin-embedded section of human intestinal cancer

IHC analysis of SLC11A1 using anti-SLC11A1 antibody (A02547-3). SLC11A1 was detected in paraffin-embedded section of mouse spleen tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-SLC11A1 Antibody (A02547-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

A02547-3 · IHCIHC analysis of SLC11A1 using anti-SLC11A1 antibody (A02547-3). SLC11A1 was detected in paraffin-embedded section of mouse spleen tissues. H

IF analysis of SLC11A1 using anti-SLC11A1 antibody (A02547-3). SLC11A1 was detected in immunocytochemical section of MCF-7 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-SLC11A1 Antibody (A02547-3) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

A02547-3 · IFIF analysis of SLC11A1 using anti-SLC11A1 antibody (A02547-3). SLC11A1 was detected in immunocytochemical section of MCF-7 cell. Enzyme anti

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000018280-SLC11A1/tissue/cerebral+cortex#imid_11755307

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000018280-SLC11A1/tissue/colon#imid_11755217

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000018280-SLC11A1/tissue/liver#imid_11755226

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000018280-SLC11A1/tissue/kidney#imid_11755227

HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA050378; green = SLC11A1, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for SLC11A1 IHC & IF

🧪Run bronchus first and look for staining in respiratory epithelial cells (HPA: Medium in bronchus respiratory epithelial cells). Use adipose tissue as the negative comparator, with adipocytes at background (HPA: Not detected in adipocytes); on the bronchus slide, score unstained neighboring cells as internal background without assuming macrophages are negative (UniProt P49279: macrophage tissue specificity).

Positive control tissue: Bronchus (Respiratory epithelial cells, HPA Medium)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SLC11A1 in HEL, MCF-7, U2OS, with annotated localisation: Mitochondria (approved) (HPA subcellular).

Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls alongside a SLC11A1 knockout specimen or a peptide-block control if the immunizing peptide is available (caption: rabbit primary; standard IHC practice). For chromogenic detection, quench endogenous peroxidase and check endogenous biotin background when using the caption’s streptavidin–biotin system (caption: SABC with DAB; standard IHC practice).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A02547-3 tissue-IHC caption does not state the fixative (caption: fixative not stated). The caption used heat retrieval in citrate buffer at pH 6 for 20 minutes, but it does not establish that this retrieval is required (caption: retrieval conditions). Neither frozen sections nor IF can be judged easier from the supplied evidence; in bronchus, check mucus and luminal material for nonspecific chromogen deposition before scoring epithelial staining (HPA: bronchus respiratory epithelial cells Medium; standard IHC practice).

HPA tissue IHC evidence for SLC11A1

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. Antibody staining in cells/structures not annotated, view images.). 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	Respiratory epithelial cells	Medium	Protein (IHC)	HPA →
Cerebral cortex	Glial cells	Medium	Protein (IHC)	HPA →
Colon	Endothelial cells	Medium	Protein (IHC)	HPA →
Duodenum	Glandular cells	Medium	Protein (IHC)	HPA →
Gallbladder	Glandular cells	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →
Bone marrow	Hematopoietic cells	Not detected	Protein (IHC)	HPA →
Caudate	Glial cells	Not detected	Protein (IHC)	HPA →
Heart muscle	Cardiomyocytes	Not detected	Protein (IHC)	HPA →
Hippocampus	Glial cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced SLC11A1 IHC Tips

Use the catalog antibody’s paraffin-section IHC conditions as a starting point, and interpret staining alongside SLC11A1’s reported cellular distribution and the controls.

Which retrieval condition should I start with for weak SLC11A1 staining?

Start with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A02547-3). The selected paraffin-section image used that condition before incubation with 1 µg/mL primary antibody overnight at 4°C (datasheet A02547-3). If staining remains weak, compare a more alkaline retrieval buffer on adjacent sections while keeping detection and exposure to chromogen matched (standard IHC practice). Include a section processed without primary antibody to distinguish retrieval-related background from specific staining (standard IHC practice). Judge improvement by staining within the expected cell population and vesicular compartment, rather than by a general increase in brown signal (UniProt P49279 localisation; standard IHC practice).

Could fixation explain inconsistent staining between paraffin sections?

The selected tissue-IHC caption identifies paraffin sections but does not report their fixative, so SLC11A1-specific fixation sensitivity is unknown (datasheet A02547-3). Record the fixative, time in fixative, processing schedule, and section age for every specimen before comparing staining (standard IHC practice). When possible, compare similarly processed sections using the same citrate pH 6, 20-minute retrieval and the same detection run (datasheet A02547-3; standard IHC practice). Include a consistent reference section in each run to detect processing or staining drift (standard IHC practice). Do not assign differences in signal to SLC11A1 abundance until those preanalytical differences have been assessed (standard IHC practice).

Should I expect membrane outlines or intracellular puncta in SLC11A1 IHC?

Prioritise intracellular vesicular staining in the appropriate cells: SLC11A1 is reported at phagosome, late-endosome, and lysosome membranes (UniProt P49279 localisation). Its 12 transmembrane segments describe an organelle membrane protein, so a crisp plasma-membrane outline alone needs independent validation (UniProt P49279 topology; standard IHC practice). Compare punctate or granular cytoplasmic signal with cell morphology and, where feasible, an adjacent section stained for a macrophage marker (UniProt P49279 tissue specificity; standard IHC practice). HPA reports variable cytoplasmic tissue staining but rates its tissue-IHC reliability Uncertain (HPA tissue IHC). Strong nuclear-only staining should therefore prompt a check of antibody specificity and detection background (UniProt P49279 localisation; standard IHC practice).

How should unknown epitope placement affect antibody validation?

SLC11A1 has 2 reported isoforms, but the supplied caption does not identify the catalog antibody’s epitope or establish isoform coverage (UniProt P49279 isoforms; datasheet A02547-3). Its N-terminal residues 1–60 and C-terminal residues 520–550 are cytoplasmic, whereas the region containing glycosylation sites 324 and 338 is annotated as extracellular in the membrane topology (UniProt P49279 topology and glycosylation). Ask for the immunogen sequence before assigning an IHC pattern to either isoform or an epitope face (standard IHC practice). Compare staining with an independently validated antibody to a distinct epitope where available (standard IHC practice). Retrieval-dependent changes alone cannot establish isoform specificity (standard IHC practice).

How can I check SLC11A1 localisation with multiplex IF?

For a follow-up IF experiment, pair SLC11A1 with a macrophage marker such as CD68 and assess signal within marker-positive cells (UniProt P49279 tissue specificity; standard IF practice). Select well-separated fluorophores and consider a far-red channel for SLC11A1 when tissue autofluorescence is prominent (standard IF practice). Because the catalog antibody’s epitope is unspecified, optimise permeabilisation on matched sections before claiming access to a cytoplasmic or organelle-lumen-facing epitope (datasheet A02547-3; UniProt P49279 topology; standard IF practice). Include single-stain and no-primary controls to assess bleed-through and background (standard IF practice). Interpret colocalisation cautiously: UniProt places SLC11A1 on phagosomal and endolysosomal membranes, while HPA reports mitochondrial and nucleolar IF localisation (UniProt P49279 localisation; HPA subcellular).

What should I adjust when DAB staining is widespread?

Compare a no-primary section with the full stain to assess background from the detection system (standard IHC practice). The selected protocol used 10% goat serum blocking, a biotinylated secondary, streptavidin–biotin detection, and DAB (datasheet A02547-3). If background persists, check the general peroxidase-blocking step, secondary-antibody binding, and endogenous biotin contribution before changing primary concentration (standard IHC practice). Shorten DAB development consistently across test sections and compare them with a matched reference section (standard IHC practice). Treat diffuse staining outside plausible cells or compartments cautiously, particularly because the available HPA tissue-IHC profile is rated Uncertain (UniProt P49279 localisation; HPA tissue IHC).

How should I quantify SLC11A1 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and tissue region before scoring, and exclude folds, necrosis, and section edges using the same rule for every specimen (standard IHC practice). Within the selected population, report percent positive cells and an H-score based on staining intensity; for dispersed positive cells, density per mm² can also be useful (standard IHC practice). Normalise cell counts to the number of evaluable cells or measured tissue area, and keep DAB development and image settings consistent (standard IHC practice). Score plausible intracellular signal separately from diffuse background, given SLC11A1’s reported vesicular localisation (UniProt P49279 localisation; standard IHC practice). Record macrophage abundance when comparing mixed tissues, since SLC11A1 is reported in macrophages (UniProt P49279 tissue specificity).

When is a positive stain convincing for SLC11A1?

A convincing result combines reproducible intracellular staining in an appropriate cell population with a clean no-primary control (UniProt P49279 tissue specificity and localisation; standard IHC practice). Check whether the signal is compatible with phagosome, late-endosome, or lysosome membranes, while recognising that chromogenic IHC cannot resolve those membranes on its own (UniProt P49279 localisation; standard IHC practice). Treat isolated nuclear staining, staining confined to section edges or necrosis, and residual endogenous-enzyme signal as possible artefacts (UniProt P49279 localisation; standard IHC practice). Do not use an HPA-positive cell type as decisive confirmation because its tissue-IHC reliability is Uncertain (HPA tissue IHC). Confirm disputed patterns with an independent antibody or complementary assay before assigning biological meaning (standard IHC practice).

Boster reagents

Best SLC11A1 / Natural resistance-associated macrophage protein 1 IHC Antibodies

A02547-3 has paraffin-section IHC images from human intestinal cancer and mouse and rat spleen, plus an IF/ICC image from MCF-7 cells (catalog image captions).

Real IHC data

Anti-NRAMP1/SLC11A1 Antibody ®

Cat # A02547-3

View product →

A02547-3 is listed for IHC and IF/ICC in human, mouse and rat samples (catalog: applications and reactivity). Its IHC captions show human intestinal cancer and mouse and rat spleen paraffin sections; its IF caption shows MCF-7 cells (catalog image captions).

Which to pick: Choose A02547-3 for paraffin-section IHC: its captions document citrate retrieval at pH 6 for 20 minutes and primary antibody at 1 μg/ml; the fixative is unreported (A02547-3 IHC captions). Choose the same SKU for IF/ICC because its application list includes both and its IF caption shows MCF-7 cells; the catalog does not report clonality. It is also the listed cross-species choice for human, mouse and rat, with paraffin-section IHC images for each species (catalog: reactivity and IHC captions).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry P49279 (NRAM1_HUMAN, Natural resistance-associated macrophage protein 1).
Human Protein Atlas. SLC11A1 tissue IHC expression (reliability: Uncertain).
Human Protein Atlas. SLC11A1 subcellular location (ICC-IF): Mainly localized to the mitochondria. In addition localized to the nucleoli..
Human Protein Atlas. SLC11A1 antibody validation summary (3 antibodies).
Peripheral Immune Cell Infiltration in the Hippocampus of Sepsis Mice. Journal of cellular and molecular medicine 2025 — PMC12808816.
SLC11A1 as a stratification indicator for immunotherapy or chemotherapy in patients with glioma. Frontiers in immunology 2022 — PMC9748290.
Influx mechanisms in the embryonic and adult rat choroid plexus: a transcriptome study. Frontiers in neuroscience 2015 — PMC4412010.
CXCL10/SLC11A1 Axis Exacerbates Septic Liver Injury by Regulating Neutrophil Extracellular Traps Formation to Drive Macrophage Pro‑Inflammatory Polarization. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2026 — PMC13360119.
PubMed PMID:7964458 — UniProt-cited evidence.
PubMed PMID:8537108 — UniProt-cited evidence.
PubMed PMID:10967134 — UniProt-cited evidence.