NUP43 / Nucleoporin Nup43 · IHC design guide

Design Immunohistochemistry for NUP43

Use colon glandular cells as a high-staining reference for NUP43 chromogenic IHC (HPA tissue IHC). Assess the chiefly cytoplasmic tissue pattern in light of NUP43’s annotated nuclear pore location (HPA tissue IHC; UniProt).

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

Immunohistochemistry protocol sheet for NUP43 (IHC for NUP43): expected localisation Chiefly cytoplasmic; nuclear in a few tissues (HPA tissue IHC), antibody A11950-1, validated IHC image, and IHC protocol steps — Printable **NUP43** IHC protocol sheet — expected localisation Chiefly cytoplasmic; nuclear in a few tissues (HPA tissue IHC), antibody A11950-1, controls and protocol steps. Open the full NUP43 IHC guide →

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NUP43 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	Chiefly cytoplasmic; nuclear in a few tissues (HPA tissue IHC)
	Staining pattern	Colon glandular cells: high, chiefly cytoplasmic staining (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet A11950-1)
	Positive control ⓘ	Adrenal gland+4 more · see all
	Negative control ⓘ	None in HPA (detected in all 45 tissues); use no-primary + isotype controls

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific)
	Caveat	Cytoplasmic IHC differs from the nuclear pore location (HPA tissue IHC; UniProt)
	Regulation	No specific expression regulator annotated (UniProt)
	Isoform / epitope	2 isoforms; epitope differences are not mapped (UniProt)

Section 1

Recommended NUP43 IHC & IF Protocols

The catalog antibody has an IHC-P protocol (datasheet A11950-1). One published study reports NUP43 IHC in mouse colorectal cancer liver metastases (PMC11102480 methods).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human colon tissue; fixative not specified (datasheet A11950-1)
Fixation	Image fixative and duration unreported (datasheet A11950-1); 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 A11950-1); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A11950-1)
Primary antibody	Rabbit anti-NUP43, 2-5 μg/ml (datasheet A11950-1)
Primary incubation	Overnight at 4 °C (datasheet A11950-1)
Detection	HRP-conjugated secondary, DAB chromogen (datasheet A11950-1)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	NUP43-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. Additional nuclear expression in a few tissues. No signal in the no-primary control.

💡Decision noteTry heat-mediated EDTA retrieval at pH 8.0 first (datasheet A11950-1); the published study does not state its retrieval conditions (PMC11102480 methods).

Section 2

What Is the Expected NUP43 Staining Pattern?

NUP43 is a nuclear pore complex and kinetochore protein with no transmembrane segment (UniProt Q8NFH3). In tissue IHC, HPA reports general cytoplasmic staining with additional nuclear staining in a few tissues, including high staining in several glandular and neuronal cell populations (HPA tissue IHC). HPA rates its tissue IHC reliability Enhanced, while noting medium agreement with RNA data and pending external verification (HPA tissue IHC).

What am I looking at on my slide?
Glandular cells in colon, duodenum or adrenal gland stain strongly; some cells also show nuclear signal (HPA tissue IHC).	This fits HPA's high staining in those cell populations and its general cytoplasmic, occasionally nuclear, tissue pattern (HPA tissue IHC). Score cytoplasmic and nuclear staining separately; nuclear staining need not appear in every tissue.
Signal appears only at the cell surface or in extracellular material, without a cellular pattern.	Treat this as suspect: NUP43 has no transmembrane segment and is assigned to the nuclear pore complex and kinetochore (UniProt Q8NFH3). Compare with a positive tissue and detection controls before assigning specificity (general IHC practice).
A cell population outside the reported high-staining groups is the strongest feature on the slide.	Possible causes include cross-reactivity or endogenous detection activity (general IHC practice). HPA's selected high-staining populations are reference examples, not an exhaustive map; its supplied negative list is empty (HPA tissue IHC).
Color spreads across stroma, empty spaces or most cells without a clear cellular pattern.	This is diffuse background rather than interpretable NUP43 localisation (general IHC practice). Review blocking, washing and chromogen development with detection controls; HPA's tissue summary does not establish a cause for background (HPA tissue IHC).
A reported high-staining population has no detectable signal, such as colon glandular cells (HPA tissue IHC).	Check tissue identity and assay performance before calling NUP43 absent (general IHC practice). HPA reports high staining in that population, but its Enhanced reliability rating still carries a pending external-verification caveat (HPA tissue IHC).

💡Expected NUP43 appearanceCall a positive result when cellular staining is clear in an HPA high-staining population, such as colon glandular cells, with the reported cytoplasmic pattern and possible additional nuclear signal; isolated extracellular or surface color is suspect (HPA tissue IHC; UniProt Q8NFH3; general IHC practice).

How each factor affects the staining
Tissue and cell choice	HPA reports high staining in adrenal, colon and duodenal glandular cells and in several neuronal populations (HPA tissue IHC). Its listed low-staining cells, such as liver cholangiocytes, are low references, not validated negative controls (HPA tissue IHC).
Compartment evidence	Tissue IHC is generally cytoplasmic with nuclear staining in a few tissues (HPA tissue IHC). UniProt assigns NUP43 to the nuclear pore complex and kinetochore (UniProt Q8NFH3); interpret a nuclear signal in the context of both records.
Antibody evidence	HPA lists Enhanced IHC validation for HPA027126, HPA027291 and HPA028500 (HPA antibodies). This supports the reported tissue pattern but does not remove HPA's medium RNA agreement and pending external-verification caveat (HPA tissue IHC).
Protein forms and topology	UniProt lists two isoforms, no signal peptide or propeptide, and no transmembrane segment (UniProt Q8NFH3). Epitope coverage is unspecified here, so these annotations cannot predict whether an antibody detects both isoforms or a particular IHC compartment.
Retrieval and detection	Antigen retrieval and endogenous detection activity are general IHC variables (general IHC practice). No NUP43-specific fixation or retrieval response is supplied; interpret changes in staining against tissue and detection controls, without attributing them to a proven NUP43 effect.
IF/ICC Q: What localisation should be compared?	A: HPA supports nucleoplasmic localisation; nuclear speckles and cytosol are marked uncertain (HPA subcellular ICC-IF). These cell-image observations provide context, while the tissue IHC pattern remains the primary guide for a chromogenic section (HPA tissue IHC).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
No stain in a reported high-staining tissue (HPA tissue IHC).	Tissue identity, retrieval or detection may need checking (general IHC practice); the record supplies no NUP43-specific fixation effect.	Verify the expected cell population, inspect a run control, then check retrieval, primary incubation and detection according to the assay controls (general IHC practice).
Strong color in a detection control without primary antibody.	Endogenous detection activity or reagent background is possible (general IHC practice).	Address the relevant endogenous activity, blocking and washing steps; reassess the tissue with matched controls (general IHC practice).
Color is diffuse and obscures cell boundaries.	Nonspecific binding, inadequate washing or excessive chromogen development may contribute (general IHC practice).	Review blocking and wash conditions, and compare a shorter development with the control slide (general IHC practice).
Only surface or extracellular staining is apparent.	That location conflicts with NUP43's annotated nuclear pore complex and kinetochore locations and lack of a transmembrane segment (UniProt Q8NFH3).	Check morphology and detection controls, then compare staining in an HPA high-staining population before interpreting the signal (HPA tissue IHC; general IHC practice).
Cytoplasmic staining is present but nuclei are unstained.	This can match HPA tissue IHC: its summary reports general cytoplasmic staining and nuclear staining in only a few tissues (HPA tissue IHC).	Record the observed compartment and cell type; do not require nuclear staining across every tissue (HPA tissue IHC).
A listed low-staining population appears intensely positive (HPA tissue IHC).	The difference warrants review, but an HPA low designation is not a validated negative result (HPA tissue IHC).	Confirm cell identification and examine primary-omission and detection controls; report the difference without assigning cross-reactivity from intensity alone (general IHC practice).

Source: UniProt Q8NFH3 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing NUP43 staining

IHC analysis of NUP43 using anti-NUP43 antibody (A11950-1). NUP43 was detected in a paraffin-embedded section of human colon tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NUP43 Antibody (A11950-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

A11950-1 · IHCIHC analysis of NUP43 using anti-NUP43 antibody (A11950-1). NUP43 was detected in a paraffin-embedded section of human colon tissue. Heat me

IF analysis of NUP43 using anti-NUP43 antibody (A11950-1) and anti-Beta Tubulin antibody (M01857-3). NUP43 was detected in immunocytochemical section of A549 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 5 μg/mL rabbit anti-NUP43 Antibody (A11950-1) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) and DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

A11950-1 · IFIF analysis of NUP43 using anti-NUP43 antibody (A11950-1) and anti-Beta Tubulin antibody (M01857-3). NUP43 was detected in immunocytochemica

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000120253-NUP43/tissue/cerebral+cortex#imid_7409288

HPA · Adrenal glandHuman Protein Atlas IHC, Adrenal gland. Source: https://www.proteinatlas.org/ENSG00000120253-NUP43/tissue/adrenal+gland#imid_7409256

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000120253-NUP43/tissue/colon#imid_7409214

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000120253-NUP43/tissue/liver#imid_7409215

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA028500; green = NUP43, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for NUP43 IHC & IF

🧪Run adrenal gland first and score glandular cells, where NUP43 staining is High (HPA: High in adrenal gland glandular cells). HPA detects NUP43 in all 45 scored tissues, so there is no HPA negative tissue; use no-primary and isotype controls, and expect any unstained cells within the positive section to remain at background rather than treating them as a validated negative cell population (HPA: no negative tissue rows; standard IHC practice).

Positive control tissue: Adrenal gland (Glandular cells, HPA High)

Negative control tissue: None in HPA: NUP43 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NUP43 in A-431, HEL, U2OS, with annotated localisation: Nucleoplasm (supported), Nuclear speckles (uncertain) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only section and a concentration-matched rabbit IgG isotype control alongside the rabbit primary antibody; confirm target dependence with knockout tissue or a peptide block if a suitable peptide is available (selected A11950-1 caption: rabbit primary; standard IHC practice). For chromogenic staining, block or assess endogenous peroxidase before interpreting DAB signal in the adrenal section (selected A11950-1 caption: HRP/DAB detection; standard IHC practice).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A11950-1 paraffin-section caption does not state the fixative (selected A11950-1 caption). That caption uses heat retrieval in EDTA at pH 8.0 for human colon; it supports a starting retrieval condition but does not establish a retrieval requirement for adrenal gland (selected A11950-1 caption). HPA has ICC-IF images in A-431, HEL and U2OS, but the supplied evidence does not establish whether IF or frozen sections are easier than paraffin IHC; assess endogenous peroxidase when interpreting adrenal DAB staining (HPA: subcellular ICC-IF images; selected A11950-1 caption: HRP/DAB; standard IHC practice).

HPA tissue IHC evidence for NUP43

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Pending external verification.). 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
Adrenal gland	Glandular cells	High	Protein (IHC)	HPA →
Bronchus	Respiratory epithelial cells	High	Protein (IHC)	HPA →
Caudate	Neuronal cells	High	Protein (IHC)	HPA →
Cerebellum	Cells in molecular layer	High	Protein (IHC)	HPA →
Cerebral cortex	Neuronal cells	High	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
None in HPA: NUP43 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.

Section 3

Advanced NUP43 IHC Tips

Troubleshoot NUP43 staining in paraffin section chromogenic IHC using the catalog antibody’s tissue method, compartment expectations and matched controls.

Which retrieval should I try first when NUP43 staining is weak?

Start with heat mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A11950-1). The selected tissue image used 2 μg/ml antibody overnight at 4°C, followed by a peroxidase conjugated secondary for 30 minutes at 37°C and DAB detection (datasheet A11950-1). If staining remains weak, compare a shorter and longer heating interval on adjacent sections while keeping antibody concentration and detection constant (standard IHC practice). Assess tissue preservation and the distribution of positive cells together, since more intense DAB alone cannot establish specific NUP43 staining (standard IHC practice).

Could fixation explain inconsistent NUP43 staining between paraffin blocks?

Target specific fixation sensitivity is unknown: the selected paraffin section caption does not state a fixative or fixation duration (datasheet A11950-1). Record fixative, fixation time and processing history for each block, then compare matched sections using the same EDTA pH 8.0 retrieval and 2 μg/ml antibody conditions (datasheet A11950-1; standard IHC practice). Include a previously stained reference block in each run to distinguish block variation from a run level change (standard IHC practice). Do not attribute a weak result to fixation alone without checking morphology, background and reference staining under the same detection conditions (standard IHC practice).

Should NUP43 staining be nuclear, cytoplasmic or at the nuclear rim?

NUP43 is a nuclear pore complex component and is also assigned to centromeres and kinetochores (UniProt Q8NFH3). Tissue IHC shows general cytoplasmic expression with additional nuclear staining in a few tissues, while cell imaging supports nucleoplasmic localization and assigns lower certainty to nuclear speckles and cytosol (HPA tissue IHC; HPA subcellular). Score nuclear and cytoplasmic DAB separately, using a nuclear counterstain and intact cells to establish compartment boundaries (standard IHC practice). A diffuse compartment pattern should be interpreted alongside a matched control and morphology, because the reported localization varies by evidence source and assay (HPA tissue IHC; HPA subcellular; standard IHC practice).

Can this antibody distinguish the two NUP43 isoforms in tissue?

The record lists 2 NUP43 isoforms, but the supplied tissue image does not identify the antibody epitope or establish isoform selectivity (UniProt Q8NFH3; datasheet A11950-1). Treat DAB staining as NUP43 associated rather than an isoform specific readout until the immunogen and both isoform sequences are compared (standard IHC practice). NUP43 has no annotated transmembrane segment or glycosylation sites, and its annotated modification includes N terminal acetylmethionine (UniProt Q8NFH3). Those features do not reveal whether the antibody recognizes both isoforms or how fixation affects its epitope; validate such claims with an appropriate independent control (standard IHC practice).

How should I investigate NUP43 localisation by multiplex IF?

Use IF as a separate validation experiment: the supplied catalog image establishes paraffin section chromogenic IHC, not IF performance (datasheet A11950-1). Pair NUP43 with a marker for the cell population being examined and a nuclear stain, then choose spectrally separated fluorophores after checking tissue autofluorescence in each channel (standard IF practice). NUP43 has no transmembrane segment and is associated with nuclear pores, kinetochores and chromosomes, so optimize permeabilisation for access to intracellular epitopes without assuming an epitope on either side of a membrane (UniProt Q8NFH3; standard IF practice). Confirm apparent nuclear or cytosolic signal with single stain and secondary only controls before comparing it with DAB patterns (standard IF practice).

What controls help identify nonspecific NUP43 DAB staining?

Check a section without primary antibody and inspect the peroxidase block, because endogenous enzyme activity or detection reagents can contribute DAB signal (standard IHC practice). Keep the selected 2 μg/ml primary incubation overnight at 4°C as the reference condition when testing lower concentrations or altered blocking (datasheet A11950-1; standard IHC practice). The selected image used 10% goat serum before primary incubation and peroxidase based DAB detection (datasheet A11950-1). Compare background in stroma, damaged regions and tissue edges with intact cells; a signal that follows tissue damage or persists without primary antibody warrants investigation before scoring (standard IHC practice).

How should I quantify NUP43 IHC across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and compartment before scoring, then report the percentage of positive cells and an H-score from intensity categories multiplied by their percentages (standard IHC practice). If counting spatially distributed positive cells instead, report density per mm² of viable tissue and keep the counting rules constant (standard IHC practice). Normalize scores to the number of evaluable cells or viable tissue area, and compare slides processed with the same retrieval, detection and counterstain settings (standard IHC practice). Report nuclear and cytoplasmic results separately, since tissue IHC describes general cytoplasmic staining with nuclear staining in a few tissues (HPA tissue IHC).

When is a NUP43 positive result convincing rather than artefactual?

A convincing result has reproducible staining in intact cells, interpretable compartment boundaries and little signal in the no primary control (standard IHC practice). Colon glandular cells are a useful positive reference because the selected paraffin image detected NUP43 in human colon and HPA reports high staining in colon glandular cells (datasheet A11950-1; HPA tissue IHC). Examine unexpected compartment staining against NUP43’s nuclear pore and kinetochore assignments and the cytoplasmic pattern reported in tissue IHC (UniProt Q8NFH3; HPA tissue IHC). Discount isolated edge staining, necrotic regions and signal consistent with residual endogenous peroxidase before assigning a biological interpretation (standard IHC practice).

Boster reagents

Best NUP43 / Nucleoporin Nup43 IHC Antibodies

Anti-NUP43 A11950-1 has human colon paraffin-section IHC images and an A549-cell IF image (catalog image captions). Listed reactivity covers human, mouse, and rat (catalog).

Real IHC data

Anti-NUP43 Antibody ®

Cat # A11950-1

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A11950-1 is shown in paraffin sections of human colon and colon adenocarcinoma using IHC (catalog IHC image captions). The same SKU is shown in A549 cells using IF (catalog IF image caption); its listed applications include IHC and ICC/IF (catalog).

Which to pick: Choose A11950-1 for paraffin-section IHC because its own images show staining in human colon and colon adenocarcinoma sections (catalog IHC image captions); the fixative is unreported (catalog IHC image captions). Choose A11950-1 for IF/ICC because its application list includes both and its IF image shows A549 cells (catalog). For mouse or rat samples, A11950-1 lists reactivity with those species, although the supplied IHC images show human tissue (catalog reactivity; catalog IHC image captions); clonality is unreported (catalog).

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

References

UniProt Consortium. UniProt entry Q8NFH3 (NUP43_HUMAN, Nucleoporin Nup43).
Human Protein Atlas. NUP43 tissue IHC expression (reliability: Enhanced).
Human Protein Atlas. NUP43 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and nuclear speckles. In addition localized to the cytosol..
Human Protein Atlas. NUP43 antibody validation summary (3 antibodies).
NUP43 promotes PD-L1/nPD-L1/PD-L1 feedback loop via TM4SF1/JAK/STAT3 pathway in colorectal cancer progression and metastatsis. Cell death discovery 2024 — PMC11102480.
Identification of a novel glycolysis-related signature to predict the prognosis of patients with breast cancer. World journal of surgical oncology 2021 — PMC8487479.
PubMed PMID:12196509 — UniProt-cited evidence.
PubMed PMID:14702039 — UniProt-cited evidence.
PubMed PMID:14574404 — UniProt-cited evidence.