SLC39A14 / Metal cation symporter ZIP14 · IHC design guide

Design Immunohistochemistry for SLC39A14

Plan chromogenic IHC on paraffin sections using the catalog antibody’s documented protocol (datasheet A04761). Kidney distal tubules and colon glandular cells provide high-staining reference populations (HPA tissue IHC).

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

Immunohistochemistry protocol sheet for SLC39A14 (IHC for SLC39A14): expected localisation Membranous and cytoplasmic staining (HPA tissue IHC), antibody A04761, validated IHC image, and IHC protocol steps — Printable **SLC39A14** IHC protocol sheet — expected localisation Membranous and cytoplasmic staining (HPA tissue IHC), antibody A04761, controls and protocol steps. Open the full SLC39A14 IHC guide →

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SLC39A14 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 and cytoplasmic staining (HPA tissue IHC)
	Staining pattern	Glandular cells: membranous and cytoplasmic staining (HPA tissue IHC)
	Antigen retrieval	Citrate pH 6 HIER, heat-mediated (datasheet A04761)
	Positive control ⓘ	Appendix+4 more · see all
	Negative control ⓘ	Adipose tissue+4 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Formaldehyde fixed in the kidney IHC example (selected-SKU IHC image A04761); keep fixation consistent (standard IHC practice; not target-specific)
	Caveat	Presumed off-target binding may complicate interpretation (HPA tissue IHC)
	Regulation	Glucose uptake enriches surface ZIP14 (UniProt)
	Isoform / epitope	3 isoforms; check extracellular vs cytoplasmic epitope (UniProt)

Section 1

Recommended SLC39A14 IHC & IF Protocols

The catalog antibody has an IHC-P protocol (datasheet: A04761). Published SLC39A14 IHC protocols cover nasopharyngeal tissue (PMC11009181) and control liver and normal human brain (PMC4894980).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Formaldehyde-fixed, paraffin-embedded human kidney tissue (datasheet A04761)
Fixation	Image formalin-fixed; duration unreported (datasheet A04761); 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 (datasheet A04761); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% serum (datasheet A04761)
Primary antibody	Rabbit anti-SLC39A14, 1-2 μg/mL (datasheet A04761)
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	SLC39A14-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Membranous and cytoplasmic expression in several tissues. No signal in the no-primary control.

💡Decision noteStart with heat-mediated citrate retrieval at pH 6 for the catalog antibody (datasheet: A04761). A published control-liver protocol used pH ~9 retrieval (PMC4894980).

Section 2

What Is the Expected SLC39A14 Staining Pattern?

SLC39A14 is a six-pass membrane transporter found at the cell surface and on endosomal and lysosomal membranes (UniProt Q15043 topology and subcellular location). In paraffin-section IHC, expect membranous and cytoplasmic staining in selected epithelial cells, including kidney distal tubules and intestinal glandular cells (HPA: tissue IHC). HPA rates the tissue evidence Enhanced, while noting medium agreement with RNA data and presumed off-target staining that was disregarded (HPA: reliability description).

What am I looking at on my slide?
Distinct membrane staining with some cytoplasmic signal in kidney distal tubules or intestinal glandular cells.	This fits the reported tissue pattern (HPA: High in kidney distal tubules and small-intestinal glandular cells; membranous and cytoplasmic profile). Score the named cell population and compartment together; intensity alone cannot establish specificity. Intracellular signal is plausible because endosome and lysosome membranes are reported locations (UniProt Q15043 subcellular location).
Predominantly nuclear staining, with little membrane or cytoplasmic signal.	Treat a nuclear-dominant pattern as discordant: the reported IHC pattern is membranous and cytoplasmic (HPA: tissue IHC profile), and the annotated locations are membranes (UniProt Q15043 subcellular location). Check the antibody and detection controls before interpreting it as SLC39A14. Neither source establishes a nuclear staining pattern.
Strong staining in a cell population listed as not detected, such as adipocytes or bone-marrow hematopoietic cells.	Investigate possible cross-reactivity or endogenous detection activity before scoring it as positive (HPA: Not detected in adipocytes and bone-marrow hematopoietic cells; general IHC practice). A not-detected reference is a comparator, not proof that every specimen or condition must be negative. Review staining in the expected cell populations on the same run.
Broad, diffuse color obscures cell borders and the tissue architecture.	Background prevents a compartment call even where SLC39A14 is expected (HPA: membranous and cytoplasmic tissue profile). Uneven reagent exposure, nonspecific binding, or detection-system background are general IHC possibilities. Compare a no-primary control and inspect whether color follows cells or spreads across the section (general IHC practice).
No detectable signal in kidney distal tubules or colon glandular cells.	A negative result in these high-staining reference populations warrants a run-level check before a biological conclusion (HPA: High in kidney distal tubules and colon glandular cells). Confirm that the positive-control section developed, then review antibody use, retrieval, and detection steps as general IHC checks. The supplied sources do not establish target-specific fixation sensitivity.

💡Expected SLC39A14 appearanceCall positive when discrete membranous with possible cytoplasmic staining marks expected cells, such as High-staining kidney distal tubules or intestinal glandular cells (HPA: tissue IHC); isolated nuclear-dominant color or staining confined to listed not-detected cells needs investigation, not automatic positive scoring (HPA: tissue IHC; UniProt Q15043 subcellular location).

How each factor affects the staining
Membrane topology and compartment	SLC39A14 has 6 transmembrane segments, extracellular and cytoplasmic regions, and reported surface, endosome, and lysosome locations (UniProt Q15043 topology and subcellular location). These support a membrane-centered interpretation with possible intracellular staining. The sources do not identify this antibody’s epitope, so topology cannot predict its retrieval requirements.
Tissue and cell selection	The tissue pattern is cell-specific: kidney distal tubules and colon, rectum, appendix, and small-intestinal glandular cells are High, while adipocytes and bone-marrow hematopoietic cells are listed as not detected (HPA: tissue IHC). Use named cell populations when comparing sections; a whole-tissue label can hide a mixed result.
Evidence strength and antibody choice	HPA calls tissue IHC Enhanced for HPA056732 and reports presumed off-target binding that was disregarded in its summary (HPA: antibody validation and reliability description). HPA016508 is ICC Approved but has no listed IHC status (HPA: antibodies). These ratings support different applications; they do not validate every unexpected staining pattern.
IF/ICC Q&A: what location should I expect?	In cultured-cell IF, HPA approves plasma-membrane, endoplasmic-reticulum, and Golgi locations and lists images from A-431, U-251MG, and U2OS (HPA: subcellular ICC-IF). This is the IF/ICC interpretation answer; it does not turn those images or the ICC approval into an IHC protocol or establish the same organelle balance in tissue sections.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Positive-control tissue is blank.	The run may have failed, or the antibody and detection conditions may be unsuitable; high-staining reference cells are expected in kidney distal tubules or colon glands (HPA: tissue IHC).	Check the control section and reagent sequence, then review retrieval and detection against the IHC-validated antibody’s instructions (general IHC practice). Do not infer an SLC39A14-specific fixation effect from this result.
Color is strongest over nuclei.	Nuclear-dominant signal conflicts with the reported membrane and cytoplasmic tissue pattern (HPA: tissue IHC profile; UniProt Q15043 subcellular location).	Inspect a no-primary control and reassess detection background and antibody specificity (general IHC practice). Require convincing staining in expected cells and compartments before scoring the section.
Color appears in adipocytes or marrow hematopoietic cells.	Those cell populations are listed as not detected; unexpected color raises a specificity or detection-background question (HPA: tissue IHC).	Compare with an expected positive tissue and a no-primary control in the same run (general IHC practice). Record the cell type and compartment rather than calling the entire tissue positive.
A uniform brown haze obscures cell outlines.	Diffuse background cannot be matched confidently to HPA’s membranous and cytoplasmic pattern (HPA: tissue IHC profile). Nonspecific detection is a general IHC possibility.	Review blocking, washes, detection exposure, and the no-primary control (general IHC practice). Score only resolved cellular staining after background is addressed.
A granular cytoplasmic pattern appears without a clear surface rim.	Intracellular membrane signal is biologically plausible because endosomes and lysosomes are annotated locations (UniProt Q15043 subcellular location), but granular color alone is not proof of target identity.	Check whether granules occur in HPA-positive cell populations and whether controls show comparable color (HPA: tissue IHC; general IHC practice). Report the observed compartment without claiming organelle identity from chromogenic IHC alone.
IF images and tissue IHC seem to emphasize different compartments.	HPA’s cultured-cell ICC-IF summary includes ER and Golgi, whereas its tissue IHC summary reports membranous and cytoplasmic expression (HPA: subcellular ICC-IF and tissue IHC).	Interpret each result within its application and cell context. Use the IF/ICC guide for that assay; the ICC Approved status of HPA016508 does not supply an IHC validation claim (HPA: antibodies).

Source: UniProt Q15043 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing SLC39A14 staining

Immunohistochemistry Validation of ZIP14 in Human Kidney Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-ZIP14 antibody (A04761) at 1 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

A04761 · IHCImmunohistochemistry Validation of ZIP14 in Human Kidney Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-ZI

Immunohistochemistry Validation of ZIP14 in Mouse Colon Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-ZIP14 antibody (A04761) at 1 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

A04761 · IHCImmunohistochemistry Validation of ZIP14 in Mouse Colon Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-ZIP1

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000104635-SLC39A14/tissue/cerebral+cortex#imid_20873087

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000104635-SLC39A14/tissue/colon#imid_20871989

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000104635-SLC39A14/tissue/liver#imid_20871990

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000104635-SLC39A14/tissue/kidney#imid_20871991

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA016508; green = SLC39A14, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for SLC39A14 IHC & IF

🧪Run kidney first: distal tubules should stain strongly (HPA: High in kidney distal tubules). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the kidney slide, compare distal tubules with adjacent cells lacking the expected distal-tubule pattern, without assuming those cells are target-free (HPA: High in distal tubules).

Positive control tissue: Appendix (Glandular cells, HPA High)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SLC39A14 in A-431, U-251MG, U2OS, with annotated localisation: Endoplasmic reticulum (approved), Golgi apparatus (approved), Plasma membrane (approved) (HPA subcellular).

Technical controls: Include a no-primary (secondary-only) slide, an irrelevant IgG control matched to the primary antibody’s host and isotype or polyclonal class, and a matched SLC39A14 knockout sample where available (standard IHC practice). In kidney chromogenic IHC, block endogenous peroxidase and check for background in the no-primary slide (standard IHC practice).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A04761 tissue-IHC metadata lists the fixative as unreported; its paraffin-section caption uses heat retrieval in citrate buffer at pH 6, but does not establish that this retrieval is required (selected A04761 tissue-IHC caption and metadata). Paraffin IHC has a supplied example, while frozen-section performance and whether IF is easier are unreported; HPA shows ICC-IF images in A-431, U-251MG and U2OS cells (selected A04761 tissue-IHC caption; HPA subcellular). For kidney sections, inspect tubules for endogenous peroxidase background in chromogenic IHC and autofluorescence if adapting the assay to IF (standard IHC/IF practice).

HPA tissue IHC evidence for SLC39A14

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
Appendix	Glandular cells	High	Protein (IHC)	HPA →
Colon	Glandular cells	High	Protein (IHC)	HPA →
Esophagus	Squamous epithelial cells	High	Protein (IHC)	HPA →
Kidney	Distal tubules	High	Protein (IHC)	HPA →
Rectum	Glandular cells	High	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 →
Breast	Adipocytes	Not detected	Protein (IHC)	HPA →
Bronchus	Respiratory epithelial cells	Not detected	Protein (IHC)	HPA →
Caudate	Glial cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced SLC39A14 IHC Tips

Troubleshoot SLC39A14 staining by checking retrieval, cell type and membrane localisation before interpreting chromogenic signal.

What retrieval should I try first for SLC39A14 in paraffin sections?

Start with heat-mediated retrieval in citrate buffer at pH 6 (datasheet A04761). Test a short and a longer heating interval on adjacent sections while holding antibody concentration, detection and development constant; the selected kidney image used 1 µg/ml primary antibody (datasheet A04761). Judge retrieval by staining in distal tubules, where expression is reported as high, alongside morphology and background (HPA: High in kidney distal tubules). If signal remains weak, evaluate an alternative retrieval buffer as a fallback, but compare it directly with citrate pH 6 and retain the condition that preserves interpretable cellular localisation (datasheet A04761; UniProt Q15043 localisation).

How can I troubleshoot weak staining when fixation varies between paraffin blocks?

Target-specific fixation sensitivity for SLC39A14 is unknown from the supplied evidence, so do not assign weak staining to a particular fixative or fixation duration. Compare blocks with documented processing histories using the same citrate pH 6 retrieval and primary concentration; the selected paraffin kidney image used 1 µg/ml antibody (datasheet A04761). Include a known positive tissue section in each run and inspect tissue integrity before adjusting retrieval or detection; distal tubules provide a reported positive cell population (HPA: High in kidney distal tubules). Record fixation history alongside staining results, and change one processing variable at a time before drawing a target-specific conclusion.

Should SLC39A14 appear only at the cell surface in IHC?

Expect a membrane-associated pattern, but assess cytoplasmic staining in context: SLC39A14 is annotated at cell, endosome and lysosome membranes (UniProt Q15043 localisation). Tissue IHC reports both membranous and cytoplasmic expression, with presumed off-target binding observed and disregarded (HPA: tissue IHC profile and reliability description). In polarised cells, inspect whether staining follows the relevant membrane domain; basolateral localisation is reported for enterocytes (UniProt Q15043; PubMed:31028174). Compare serial sections and a no-primary control before treating diffuse cytoplasmic colour as specific, and document membrane and intracellular scores separately when both patterns are present (HPA: tissue IHC profile; UniProt Q15043 localisation).

Could epitope placement explain inconsistent SLC39A14 staining across sections?

Yes, but the supplied evidence does not identify the catalog antibody’s epitope, so its isoform coverage and accessibility cannot be assigned. SLC39A14 has 3 annotated isoforms, a cleaved 1–30 signal sequence, and 6 transmembrane segments (UniProt Q15043 isoforms, processing and topology). An extracellular epitope and a cytoplasmic epitope may respond differently to section processing; the large cytoplasmic interval spans residues 246–397 (UniProt Q15043 topology). Request epitope or immunogen information for the exact antibody, then compare staining under the validated citrate pH 6 retrieval condition before interpreting differences as biological isoform expression (datasheet A04761; UniProt Q15043 isoforms).

How should I follow up an IHC pattern with multiplex immunofluorescence?

Use IF as a separate validation experiment and pair SLC39A14 with a marker that identifies the expected cell population; kidney distal tubules are a reported high-staining population (HPA: High in kidney distal tubules). Choose spectrally separated fluorophores after checking unstained-section autofluorescence, then acquire single-stain controls before interpreting overlap. Set permeabilisation according to the antibody’s documented epitope: SLC39A14 has extracellular and cytoplasmic regions, including residues 246–397 on the cytoplasmic side (UniProt Q15043 topology). If the epitope is unknown, compare otherwise matched permeabilised and non-permeabilised sections, and do not transfer the paraffin IHC antibody concentration of 1 µg/ml to IF without optimisation (datasheet A04761).

What should I check when SLC39A14 staining is diffuse or widespread?

First examine a no-primary control and the section edges for detection background, then check whether signal tracks intact cells rather than folds, debris or damaged regions. For chromogenic detection, include an endogenous peroxidase block and assess its effect before attributing brown colour to SLC39A14. Titrate around the selected kidney image’s 1 µg/ml primary concentration, keeping citrate pH 6 retrieval and development conditions fixed during that comparison (datasheet A04761). Interpret widespread staining against the reported membranous and cytoplasmic tissue profile, while remembering that presumed off-target binding was observed in the tissue survey (HPA: tissue IHC profile and reliability description).

How should I score SLC39A14 staining across samples? ⚠ ANSWER MARKED FOR VERIFICATION

Define the scored cell population before reviewing outcomes; distal tubules are a reported high-staining population in kidney (HPA: High in kidney distal tubules). For chromogenic IHC, record the percentage of positive cells and an H-score using intensity categories 0–3, and score membrane and cytoplasmic staining separately when both are present (HPA: tissue IHC profile). Normalise counts to the number of evaluable cells in the same compartment, or express positive-cell density per mm² of viable tissue when cell counting is impractical. Hold retrieval, antibody concentration, imaging and scoring thresholds constant across samples; citrate pH 6 and 1 µg/ml are the selected image’s documented conditions (datasheet A04761).

How can I distinguish true SLC39A14 staining from artefact?

Look for reproducible staining in intact, expected cells and plausible compartments: kidney distal tubules stain highly, and SLC39A14 is annotated at cell and intracellular membranes (HPA: High in kidney distal tubules; UniProt Q15043 localisation). Be cautious with nuclear-only colour, abrupt section-edge enhancement, necrotic areas or signal that also appears without primary antibody; these patterns warrant control review before scoring. Check endogenous peroxidase blocking when diffuse chromogen persists, and compare adjacent sections processed with the same citrate pH 6 retrieval (datasheet A04761). Treat a plausible pattern as supportive rather than definitive, because the tissue IHC survey reports medium staining–RNA consistency and presumed off-target binding (HPA: reliability description).

Boster reagents

Best SLC39A14 / Metal cation symporter ZIP14 IHC Antibodies

The IHC-validated antibody A04761 has paraffin-section images from human kidney, mouse colon and rat kidney (A04761 image captions); no IF/ICC validation is listed (A04761 applications).

Real IHC data

Anti-ZIP14 SLC39A14 Antibody

Cat # A04761

View product →

A04761 is listed for IHC-P in human, mouse and rat samples (A04761 applications and reactivity). Its images show paraffin sections of human kidney, mouse colon and rat kidney; the captions report formaldehyde fixation (A04761 image captions).

Which to pick: Choose A04761 for paraffin-section IHC: it is a rabbit polyclonal antibody listed for IHC-P, with 1–2 μg/mL as the catalog starting range (A04761 catalog entry and datasheet). For cross-species IHC, its captions show human kidney and mouse colon at 1 μg/mL and rat kidney at 2 μg/mL, all in paraffin sections fixed with formaldehyde (A04761 image captions). No IF/ICC application or figure is listed for A04761, so the payload does not support an IF/ICC pick (A04761 applications and image captions).

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

References

UniProt Consortium. UniProt entry Q15043 (S39AE_HUMAN, Metal cation symporter ZIP14).
Human Protein Atlas. SLC39A14 tissue IHC expression (reliability: Enhanced).
Human Protein Atlas. SLC39A14 subcellular location (ICC-IF): Localized to the plasma membrane, the Golgi apparatus and endoplasmic reticulum..
Human Protein Atlas. SLC39A14 antibody validation summary (2 antibodies).
Decreased expression of SLC 39A14 is associated with tumor aggressiveness and biochemical recurrence of human prostate cancer. OncoTargets and therapy 2016 — PMC4948725.
Integrating iron metabolism-related gene signature to evaluate prognosis and immune infiltration in nasopharyngeal carcinoma. Discover oncology 2024 — PMC11009181.
Mutations in SLC39A14 disrupt manganese homeostasis and cause childhood-onset parkinsonism-dystonia. Nature communications 2016 — PMC4894980.
Behavioral and neurochemical studies of inherited manganese-induced dystonia-parkinsonism in Slc39a14-knockout mice. Neurobiology of disease 2021 — PMC8448284.
PubMed PMID:7584044 — UniProt-cited evidence.
PubMed PMID:14702039 — UniProt-cited evidence.
PubMed PMID:16421571 — UniProt-cited evidence.