NME1 / Nucleoside diphosphate kinase A · IHC design guide

Design Immunohistochemistry for NME1

Plan chromogenic NME1 IHC in paraffin sections around the predominantly cytoplasmic tissue pattern (HPA tissue IHC). Breast glandular cells and lymph node non-germinal center cells show high staining, while muscle cells are reported as not detected (HPA tissue IHC).

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

Immunohistochemistry protocol sheet for NME1 (IHC for NME1): expected localisation Predominantly cytoplasmic in tissues (HPA tissue IHC), antibody PA1829, validated IHC image, and IHC protocol steps — Printable **NME1** IHC protocol sheet — expected localisation Predominantly cytoplasmic in tissues (HPA tissue IHC), antibody PA1829, controls and protocol steps. Open the full NME1 IHC guide →

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NME1 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	Predominantly cytoplasmic in tissues (HPA tissue IHC)
	Staining pattern	Cytoplasmic staining in most tissue cell types (HPA tissue IHC)
	Antigen retrieval	Citrate pH 6 HIER, heat-mediated (datasheet PA1829)
	Positive control ⓘ	Breast+4 more · see all
	Negative control ⓘ	Heart muscle+2 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep fixation conditions consistent across samples (standard IHC practice; not target-specific)
	Caveat	DNA damage can shift NME1 into nuclei (UniProt)
	Regulation	Ubiquitously expressed (UniProt)
	Isoform / epitope	3 isoforms; epitope coverage needs verification (UniProt)

Section 1

Recommended NME1 IHC & IF Protocols

The catalog antibody’s IHC-P protocol is followed by three published NME1 protocols for lung, colorectal, and mouse liver sections (PMC7598190; PMC5622428; PMC6376988).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded Rat Cerebellum tissues; fixative not specified (datasheet PA1829)
Fixation	Image fixative and duration unreported (datasheet PA1829); 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 PA1829)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet PA1829)
Primary antibody	Rabbit anti-NME1, 0.5-1μg/ml (datasheet PA1829)
Primary incubation	Overnight at 4 °C (datasheet PA1829)
Detection	Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PA1829)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	NME1-positive staining in glandular cells of breast (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control.

💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet PA1829); the lung study used citrate at pH 6 (PMC7598190). The colorectal study used conditioner at pH 8 (PMC5622428).

Section 2

What Is the Expected NME1 Staining Pattern?

NME1 staining should be mainly cytoplasmic in most tissues, with a supported cytosolic IF pattern (HPA tissue IHC; HPA subcellular). Breast glandular cells and lymph node non-germinal center cells are useful high-staining examples (HPA: High). UniProt also lists nuclear and cell-membrane localization, so compartment calls need context; NME1 has no transmembrane segment (UniProt P15531 localization and topology). HPA rates the tissue IHC pattern Supported, with medium consistency between staining and RNA data (HPA tissue IHC).

What am I looking at on my slide?
Cytoplasmic stain in breast glandular cells or lymph node non-germinal center cells.	This matches the predominant compartment and two high-staining cell populations (HPA: cytoplasmic in most tissues; High in these cells). Score the named cells, since nearby cell populations may differ (general IHC practice).
Predominantly nuclear staining, or a sharp membrane rim with little cytoplasmic signal.	Recheck controls and compartment scoring: HPA reports mainly cytosolic IF, while UniProt permits nuclear and cell-membrane localization (HPA subcellular; UniProt P15531). Nuclear signal alone cannot establish an artefact; a uniformly rim-only pattern is atypical of the predominant HPA pattern.
Strong stain in cardiomyocytes, skeletal myocytes, or smooth muscle cells.	HPA lists these specific cells as Not detected (HPA tissue IHC). Check cell identity and controls before interpreting the signal as NME1; cross-reactivity or endogenous chromogen activity are assay possibilities (general IHC practice).
Diffuse stain across tissue and empty spaces, with poor cell boundaries.	This does not resemble the reported cellular cytoplasmic pattern (HPA tissue IHC). Broad background can arise from nonspecific reagent binding or detection chemistry; inspect a no-primary control and blocking steps (general IHC practice).
No stain in breast glandular cells on a candidate positive section.	The result conflicts with an HPA High example (HPA: breast glandular cells). Verify that the expected cells are present, then assess the assay with matched controls; absence on one section does not establish biological loss (general IHC practice).

💡Expected NME1 appearanceCall positive a mainly cytoplasmic stain in the expected cells, especially High breast glandular or lymph node non-germinal center cells; diffuse stain outside cell boundaries or strong signal in HPA Not detected muscle cells calls for control review (HPA tissue IHC; general IHC practice).

How each factor affects the staining
Tissue and cell selection	NME1 has low tissue RNA specificity and cytoplasmic protein staining in most tissues, but cell-level IHC varies: breast glandular cells and lymph node non-germinal center cells are High, while the three listed muscle cell types are Not detected (HPA tissue IHC).
Compartment interpretation	HPA IF places NME1 mainly in the cytosol; UniProt also lists nucleus and cell membrane, including cell-cycle-dependent nuclear localization and translocation after DNA damage (HPA subcellular; UniProt P15531). Use the predominant pattern without treating every nuclear signal as invalid.
Antibody evidence	The supplied HPA antibodies have IHC Supported status; that validation supports use of the observed pattern but does not prove every positive cell or exclude sample-specific background (HPA antibody validation; general IHC practice). Interpret the chosen antibody with section-level controls.
Protein forms and epitope limits	UniProt lists 3 isoforms, no signal peptide or transmembrane segment, and a chain spanning residues 2–152 (UniProt P15531). Without an antibody epitope or isoform-reactivity record, these facts cannot predict which forms the stain detects.
IF/ICC: what pattern should be expected?	Mainly cytosolic signal is the supported HPA IF result (HPA subcellular). This is a compartment cross-check for IHC interpretation; the supplied IF record does not specify an IF antibody setup or protocol (HPA subcellular).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Expected positive cells are unstained.	A failed IHC workflow is possible; HPA reports High staining in breast glandular cells and lymph node non-germinal center cells (HPA tissue IHC; general IHC practice).	Confirm the cell type and examine a matched known-positive section. Review antibody dilution, detection reagents, and the retrieval procedure used for that antibody (general IHC practice); no NME1-specific retrieval condition is supplied.
Muscle cells stain strongly.	Cardiomyocytes, skeletal myocytes, and smooth muscle cells are listed as Not detected (HPA tissue IHC). Cross-reactivity or endogenous detection activity could explain unexpected stain (general IHC practice).	Check morphology and a no-primary control; review endogenous enzyme blocking if using an enzyme-based chromogen (general IHC practice). Reassess the call against the HPA cell-specific pattern.
Brown signal coats spaces or tissue edges.	A broad deposit lacks the cellular boundaries expected for cytoplasmic NME1 staining (HPA tissue IHC). Detection precipitate or nonspecific background is possible (general IHC practice).	Inspect no-primary and reagent controls, wash steps, and chromogen development; score only signal assignable to cells (general IHC practice).
Nuclear stain dominates the field.	UniProt permits nuclear localization, but HPA describes mainly cytosolic IF and cytoplasmic staining in most tissues (UniProt P15531; HPA subcellular; HPA tissue IHC).	Compare the cytoplasmic signal and controls before scoring. Record nuclear staining separately with its cell context; do not label it an artefact solely because it is nuclear (UniProt P15531; general IHC practice).
Every cell appears equally dark.	HPA reports differing cell-level intensities, including High, Medium, Low, and Not detected examples (HPA tissue IHC). Uniform dark staining may obscure that pattern through background or overdevelopment (general IHC practice).	Compare a no-primary control and review detection development. Score named cell populations rather than whole-section darkness (general IHC practice).
IF looks cytosolic while IHC also shows nuclear signal.	These observations can coexist: HPA supports mainly cytosolic IF, and UniProt lists context-dependent nuclear localization (HPA subcellular; UniProt P15531).	Use IF as a compartment comparison, then evaluate IHC cell identity, morphology, and controls on its own terms (general IHC practice). Do not infer an IF protocol from this IHC result.

Source: UniProt P15531 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing NME1 staining

IHC analysis of NM23A using anti-NM23A antibody (PA1829). NM23A was detected in paraffin-embedded section of Rat Cerebellum 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-NM23A Antibody (PA1829) 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.

PA1829 · IHCIHC analysis of NM23A using anti-NM23A antibody (PA1829). NM23A was detected in paraffin-embedded section of Rat Cerebellum tissues. Heat me

IHC analysis of NM23A using anti-NM23A antibody (PA1829). NM23A was detected in immunocytochemical section of HELA 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 1μg/ml rabbit anti-NM23A Antibody (PA1829) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

PA1829 · IHCIHC analysis of NM23A using anti-NM23A antibody (PA1829). NM23A was detected in immunocytochemical section of HELA Cell. Enzyme antigen retr

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000239672-NME1/tissue/cerebral+cortex#imid_10276861

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000239672-NME1/tissue/colon#imid_10276787

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000239672-NME1/tissue/liver#imid_10276788

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000239672-NME1/tissue/kidney#imid_10276797

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB040571; green = NME1, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for NME1 IHC & IF

🧪Run breast first: its glandular cells should stain strongly (HPA: High in breast glandular cells). Use heart muscle cardiomyocytes as the negative tissue (HPA: Not detected in cardiomyocytes); compare nonglandular cells on the breast slide for background, but do not assume they lack NME1 (UniProt P15531: ubiquitously expressed).

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

Negative control tissue: Heart muscle (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NME1 in A-431, U-251MG, U2OS, HaCaT, SiHa, Sperm, with annotated localisation: Cytosol (supported) (HPA subcellular).

Technical controls: Include no-primary (secondary-only), a host-matched rabbit IgG isotype control, and NME1-knockout material as a biological negative (PA1829 caption: rabbit primary; standard IHC practice). For breast sections, block endogenous peroxidase and assess endogenous biotin background when using the caption’s biotin-based DAB detection (PA1829 caption: biotinylated secondary, SABC and DAB; standard IHC practice).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected PA1829 paraffin-section caption does not state a fixative (PA1829 caption: fixative not stated). That caption uses citrate retrieval at pH 6 for 20 minutes, but does not establish that retrieval is required; HPA ICC-IF images support cytosolic detection, without establishing whether frozen sections or IF are easier (PA1829 caption; HPA: Cytosol supported). No breast-specific artefact is documented in the supplied evidence; check background with the stated controls (HPA: breast glandular cells High; standard IHC practice).

HPA tissue IHC evidence for NME1

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium 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
Breast	Glandular cells	High	Protein (IHC)	HPA →
Endometrium	Glandular cells	Medium	Protein (IHC)	HPA →
Lymph node	Non-germinal center cells	High	Protein (IHC)	HPA →
Adrenal gland	Glandular cells	Medium	Protein (IHC)	HPA →
Appendix	Glandular cells	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Heart muscle	Cardiomyocytes	Not detected	Protein (IHC)	HPA →
Skeletal muscle	Myocytes	Not detected	Protein (IHC)	HPA →
Smooth muscle	Smooth muscle cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced NME1 IHC Tips

Troubleshoot NME1 staining in paraffin section IHC by checking retrieval, cellular distribution, controls, and scoring before interpreting biological differences.

How should I retrieve NME1 in paraffin sections when staining is weak?

Start with heat-mediated citrate retrieval at pH 6 for paraffin sections (datasheet PA1829). The selected tissue IHC image used 20 minutes of heating in that buffer, followed by 1 µg/mL primary antibody overnight at 4°C (caption PA1829). When staining is weak, compare a small retrieval time series while keeping section thickness, antibody concentration, and detection constant (standard IHC practice). Check tissue integrity alongside signal: excessive heating can damage morphology, making cell-level interpretation unreliable (standard IHC practice). Include a concurrently processed positive tissue section to distinguish a retrieval problem from a failed staining run (standard IHC practice).

Could fixation explain inconsistent NME1 staining between paraffin blocks?

The selected paraffin section caption does not report its fixative, and target-specific fixation sensitivity for NME1 is unknown (caption PA1829). Record each block's fixative and fixation duration, then compare blocks using the same pH 6 citrate retrieval and detection conditions (datasheet PA1829; standard IHC practice). Include a known staining control in every run and assess morphology before attributing an intensity difference to NME1 abundance (standard IHC practice). If blocks differ in processing history, optimize retrieval on representative sections and document the chosen conditions for each group (standard IHC practice). Do not infer a fixation effect from tissue staining patterns alone (standard IHC practice).

Should NME1 staining be cytoplasmic, nuclear, or membranous?

Expect predominantly cytoplasmic staining in tissue sections, consistent with the supported cytosolic location in cell imaging (HPA tissue IHC; HPA subcellular). Nuclear NME1 is biologically plausible because its localization can change with the cell cycle or DNA damage, while UniProt also lists the cell membrane (UniProt P15531). Because NME1 has no transmembrane segment, a sharp membrane rim needs corroboration before being interpreted as membrane-associated protein (UniProt P15531 topology; standard IHC practice). Score cytoplasmic and nuclear signal separately and compare each pattern with counterstained cell morphology (standard IHC practice). Review unusual compartment staining against omission controls and a second validated assay where available (standard IHC practice).

How can epitope uncertainty affect NME1 staining across samples?

NME1 has 3 annotated isoforms, but the supplied product caption does not define the antibody epitope or its isoform coverage (UniProt P15531; caption PA1829). UniProt also annotates modified residues, including lysine 56, cysteine 109, and serine 120; these annotations alone do not establish an effect on this antibody's staining (UniProt P15531). Obtain the immunogen or epitope information before claiming that a staining difference reflects an isoform or modification (standard IHC practice). If an epitope is mapped, check which isoforms contain it and whether retrieval conditions preserve interpretable morphology (standard IHC practice). Treat unexplained staining differences as provisional until orthogonal evidence supports their identity (standard IHC practice).

How should I investigate NME1 localisation by multiplex IF?

Use IF to examine intracellular distribution alongside a cell-type marker validated for the tissue under study; glandular cells in breast are a documented NME1-positive reference (HPA tissue IHC; standard IF practice). Select a fluorophore in a spectral channel with low measured tissue autofluorescence, and acquire unstained and single-channel controls before interpreting overlap (standard IF practice). For a cytosolic or nuclear epitope, test gentle permeabilisation after fixation, since NME1 is mainly cytosolic and has no transmembrane segment (HPA subcellular; UniProt P15531 topology; standard IF practice). Compare permeabilised and unpermeabilised samples while keeping acquisition settings fixed (standard IF practice). Verify antibody performance for IF separately from its paraffin section IHC use (standard IF practice).

What causes diffuse brown background in NME1 IHC?

First compare the stained section with a no-primary control and inspect whether color follows cells, section edges, or debris (standard IHC practice). The selected method uses a biotinylated secondary, streptavidin–biotin detection, and DAB, so check secondary-only signal and endogenous biotin when background is widespread (caption PA1829; standard IHC practice). Include a peroxidase block before DAB development and verify that washing removes loosely bound reagent; these are general chromogenic workflow steps (standard IHC practice). Titrate the primary around the reported 1 µg/mL condition while holding development time constant (caption PA1829; standard IHC practice). Assess background before scoring because NME1 is reported across many tissues (UniProt P15531).

How should I score NME1 across sections with different cell mixtures? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and compartment before scoring; HPA reports cytoplasmic expression in most tissues and cell-dependent staining levels (HPA tissue IHC). For a chosen population, record the percentage of positive cells and staining intensity, then calculate an H-score from the same intensity bins across samples (standard IHC practice). Normalize positive-cell counts to the number of eligible cells, or report positive-cell density per mm² of viable tissue when cell counts are impractical (standard IHC practice). Score nuclear and cytoplasmic staining separately because both locations are annotated for NME1 (UniProt P15531; standard IHC practice). Keep retrieval, DAB development, image settings, and exclusion criteria consistent across the comparison (standard IHC practice).

When is an unexpected NME1-positive area likely to be artefact?

Start by identifying the stained cells and compartment: predominantly cytoplasmic tissue staining is expected, although nuclear localization is documented under specific conditions (HPA tissue IHC; UniProt P15531). High staining in breast glandular cells or lymph node non-germinal center cells is a useful reference, whereas cardiomyocytes and skeletal myocytes were not detected in the HPA tissue set (HPA tissue IHC). Treat signal confined to section edges, necrotic material, or no-primary controls as suspect and repeat with matched processing (standard IHC practice). Check endogenous peroxidase when brown DAB signal appears outside plausible cells (standard IHC practice). Confirm surprising patterns with independent specificity evidence before assigning biological meaning (standard IHC practice).

Boster reagents

Best NME1 / Nucleoside diphosphate kinase A IHC Antibodies

PA1829 has real IHC images from rat cerebellum and human mammary cancer tissue; M01334 lists human IHC and IF applications but has no image captions (PA1829 IHC image captions; M01334 catalog).

Real IHC data

Anti-NM23A/NME1 Antibody ®

Cat # PA1829

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PA1829 has paraffin-section IHC images from rat cerebellum and human mammary cancer tissue, plus a chromogenic immunocytochemistry image of HeLa cells (PA1829 IHC image captions). M01334 lists human IHC, IF and ICC applications, but has no IHC or IF image captions (M01334 catalog applications and image alts).

Which to pick: For tissue IHC, choose PA1829: its captions document paraffin sections with citrate pH 6 retrieval, though the fixative is unreported (PA1829 IHC image captions). For human IF/ICC, M01334 is a rabbit monoclonal listed for both applications, but has no fluorescence image; PA1829 lists human ICC and has a chromogenic HeLa cell image (M01334 catalog and image alts; PA1829 catalog and IHC image captions). For cross-species IHC, PA1829 lists human, mouse and rat reactivity, with tissue images from human and rat; M01334 lists human reactivity only (PA1829 catalog and IHC image captions; M01334 catalog).

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

References

UniProt Consortium. UniProt entry P15531 (NDKA_HUMAN, Nucleoside diphosphate kinase A).
Human Protein Atlas. NME1 tissue IHC expression (reliability: Supported).
Human Protein Atlas. NME1 subcellular location (ICC-IF): Mainly localized to the cytosol..
Human Protein Atlas. NME1 antibody validation summary (6 antibodies).
Negative Effect of Reduced NME1 Expression on Recurrence-Free Survival in Early Stage Non-Small Cell Lung Cancer. Journal of clinical medicine 2020 — PMC7598190.
Clinical Implication and the Hereditary Factors of NM23 in Hepatocellular Carcinoma Based on Bioinformatics Analysis and Genome-Wide Association Study. Journal of oncology 2018 — PMC6312618.
A transcriptome-based protein network that identifies new therapeutic targets in colorectal cancer. BMC genomics 2017 — PMC5622428.
The protein histidine phosphatase LHPP is a tumour suppressor. Nature 2018 — PMC6376988.
PubMed PMID:2509941 — UniProt-cited evidence.
PubMed PMID:1851158 — UniProt-cited evidence.
PubMed PMID:7916650 — UniProt-cited evidence.