NME2 / Nucleoside diphosphate kinase B · IHC design guide

Design Immunohistochemistry for NME2

Plan NME2 staining in paraffin sections using the catalog antibody and the reported cytoplasmic tissue pattern (HPA tissue IHC). Kidney tubule cells show high staining, while adipocytes are reported as not detected; interpret results with the atlas warning that staining may reflect more than one gene (HPA tissue IHC).

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

Immunohistochemistry protocol sheet for NME2 (IHC for NME2): expected localisation General cytoplasmic staining in tissue (HPA tissue IHC), antibody PA1830, validated IHC image, and IHC protocol steps — Printable **NME2** IHC protocol sheet — expected localisation General cytoplasmic staining in tissue (HPA tissue IHC), antibody PA1830, controls and protocol steps. Open the full NME2 IHC guide →

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NME2 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	General cytoplasmic staining in tissue (HPA tissue IHC)
	Staining pattern	General cytoplasmic staining (HPA tissue IHC)
	Antigen retrieval	Citrate pH 6 HIER, heat-mediated (datasheet PA1830)
	Positive control ⓘ	Endometrium+4 more · see all
	Negative control ⓘ	Adipose tissue+3 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific)
	Caveat	Staining may reflect proteins from more than one gene (HPA tissue IHC)
	Regulation	Expression regulation unreported (UniProt)
	Isoform / epitope	2 isoforms (1, 3); epitope impact unknown (UniProt)

Section 1

Recommended NME2 IHC & IF Protocols

The catalog antibody’s IHC-P protocol (datasheet: PA1830) is accompanied by published NME2 IHC parameters from three articles (PMC4336288; PMC12150935; PMC8572706).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Tissue sections; selected-image fixative not specified (standard IHC workflow)
Fixation	Image fixative and duration unreported (datasheet PA1830); 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 PA1830)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet PA1830)
Primary antibody	Rabbit anti-NME2, 0.5-1μg/ml (datasheet PA1830)
Primary incubation	Overnight at 4 °C (datasheet PA1830)
Detection	Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PA1830)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	NME2-positive staining in glandular cells of endometrium (HPA tissue IHC: Medium). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control.

💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet: PA1830); the rat skin protocol also reports this condition (PMC8572706).

Section 2

What Is the Expected NME2 Staining Pattern?

NME2 staining should be mainly cytoplasmic, with nuclear and peripheral cell-projection localisation also reported (HPA: general cytoplasmic IHC profile; UniProt P22392 localisation). Strong staining is reported in kidney tubules, urothelium and several glandular epithelia (HPA: High in those cells). NME2 has no transmembrane segment (UniProt P22392 topology). Interpret the pattern cautiously: HPA rates tissue IHC Approved, with medium RNA–staining consistency, pending external verification and a warning that the antibody targets protein from more than one gene.

What am I looking at on my slide?
Cytoplasmic staining in kidney tubular cells, urothelium or prostate glandular cells.	This fits the reported compartment and high-staining cell types (HPA: general cytoplasmic profile; High in kidney tubules, urothelium and prostate glands). Score the relevant cells separately from neighboring tissue; staining alone does not establish that every detected molecule is NME2 (HPA: antibody targets protein from more than one gene).
Predominantly crisp cell-surface or extracellular staining, with little cytoplasmic signal.	An exclusively surface or extracellular pattern does not fit the expected general cytoplasmic IHC profile (HPA: tissue IHC profile). NME2 has no transmembrane segment, although UniProt also reports localisation at peripheral ruffles and lamellipodia (UniProt P22392 topology and localisation). Review the signal as a possible artefact before calling it positive.
Strong staining in adipocytes or muscle cells while the expected positive cells stain weakly.	HPA reports NME2 as Not detected in adipocytes, cardiomyocytes, skeletal myocytes and smooth muscle cells (HPA: tissue IHC). Investigate cross-reactivity or endogenous chromogenic activity (HPA: antibody targets protein from more than one gene; general IHC practice). These cells are comparison points, not proof of universal biological absence.
Diffuse brown haze across cells, stroma and the section background.	A haze without cell-specific localisation cannot be scored as the reported cytoplasmic pattern (HPA: tissue IHC profile). Check nonspecific detection and background with appropriate controls, then reassess blocking and washing (general IHC practice). Do not infer NME2 abundance from background intensity.
No staining in kidney tubular cells or another reported high-staining cell population.	The result conflicts with an HPA high-staining example, but a single blank section cannot distinguish technical failure from sample variation (HPA: High in kidney tubules; Approved IHC with medium RNA–staining consistency). Check section quality, the validated IHC workflow and a control section before interpreting it as absent expression (general IHC practice).

💡Expected NME2 appearanceCall an IHC section positive when the relevant cells show distinct, mainly cytoplasmic signal—potentially strong in kidney tubules or urothelium (HPA: general cytoplasmic profile; High in both cell types)—while diffuse haze or an exclusively extracellular deposit is treated as suspect (HPA: tissue IHC profile; UniProt P22392 localisation; general IHC practice).

How each factor affects the staining
Which compartment is expected?	HPA describes mainly cytoplasmic tissue staining, while UniProt also lists nucleus, perinuclear cytoplasm, ruffles and lamellipodia (HPA: tissue IHC profile; UniProt P22392 localisation). A nuclear component can therefore be plausible; an isolated nuclear pattern needs review against the cytoplasmic IHC profile.
Which tissue comparisons are informative?	Kidney tubules, lymph-node non-germinal-center cells, respiratory epithelium, several glandular epithelia and urothelium are reported High; adipocytes and muscle cells are Not detected (HPA: tissue IHC). Compare defined cell populations, since UniProt describes NME2 as ubiquitously expressed (UniProt P22392 tissue specificity).
How certain is antibody-specific attribution?	HPA rates tissue IHC Approved but reports medium consistency with RNA data, pending external verification, and detection of protein from more than one gene (HPA: tissue IHC reliability). Its antibody list assigns IHC Approved to HPA041113; ICC support for other listed antibodies does not establish their IHC performance (HPA: antibody validation).
Do sequence annotations predict a retrieval setting?	Two isoforms are listed, with no annotated signal peptide, propeptide, glycosylation sites or modified residues (UniProt P22392 isoforms and processing). The supplied evidence gives no epitope position or target-specific fixation sensitivity, so it cannot justify an NME2-specific retrieval condition; follow the antibody's validated IHC instructions (general IHC practice).
IF/ICC Q&A: what localisation should an IF image show?	Mainly cytosolic fluorescence is the HPA-supported ICC-IF result (HPA: Cytosol, supported). UniProt also reports nuclear and peripheral localisation (UniProt P22392 localisation). Assess an IF image using its own antibody and controls; HPA's supported ICC antibodies are listed separately from its IHC Approved antibody (HPA: antibody validation).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
A reported high-staining tissue is blank.	The section or detection workflow may have failed; HPA's High designation is an observation, not a guarantee for every specimen (HPA: tissue IHC; general IHC practice).	Run a known-positive section alongside it and check the antibody-specific IHC instructions, reagent activity and detection steps (general IHC practice).
Most compartments have uniform brown background.	Nonspecific binding, residual detection reagent or endogenous peroxidase activity can obscure cellular staining in chromogenic IHC (general IHC practice).	Inspect a no-primary control; review blocking, washing and endogenous-peroxidase control steps before scoring (general IHC practice).
A sharp membrane rim is the only signal.	An exclusively membrane-bound pattern conflicts with the general cytoplasmic profile and the lack of a transmembrane segment (HPA: tissue IHC profile; UniProt P22392 topology).	Check the positive control for cytoplasmic signal and compare the rim with background and no-primary controls (general IHC practice).
Adipocytes or muscle cells stain more strongly than nearby expected-positive cells.	This reverses HPA's reported Not detected pattern in those cell types; off-pattern signal may include cross-reactivity or endogenous chromogenic activity (HPA: tissue IHC and antibody reliability; general IHC practice).	Identify the stained cell type, inspect no-primary controls and seek agreement with an independently validated antibody before attributing the signal to NME2 (general IHC practice).
Only nuclei stain strongly.	Nuclear localisation is reported by UniProt, but HPA describes general tissue staining as cytoplasmic (UniProt P22392 localisation; HPA: tissue IHC profile).	Keep the nuclear observation distinct from the expected cytoplasmic score; compare with a reported high-staining tissue and review counterstain and controls (HPA: tissue IHC; general IHC practice).
The staining pattern varies across antibody preparations.	Antibody-specific recognition can differ, and HPA cautions that its tissue IHC antibody targets protein from more than one gene (HPA: antibody reliability).	Record the antibody identity and validation status, then compare the same cell populations under controlled IHC conditions (HPA: antibody validation; general IHC practice).

Source: UniProt P22392 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing NME2 staining

PA1830 · IHCAnti-NME2 antibody, PA1830, IHC(P) IHC(P): Human Intestinal Cancer Tissue

PA1830 · IHCAnti-NME2 antibody, PA1830, IHC(P) IHC(P): Rat Cerebellum Tissue

IF analysis of NME2 using anti-NME2 antibody (PA1830). NME2 was detected in immunocytochemical section of U20S cells. 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-NME2 Antibody (PA1830) overnight at 4°C. DyLight®550 Conjugated Goat Anti-Rabbit IgG (BA1135) 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.

PA1830 · IFIF analysis of NME2 using anti-NME2 antibody (PA1830). NME2 was detected in immunocytochemical section of U20S cells. Enzyme antigen retriev

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000243678-NME2/tissue/cerebral+cortex#imid_13092147

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000243678-NME2/tissue/colon#imid_13092065

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000243678-NME2/tissue/liver#imid_13092082

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000243678-NME2/tissue/kidney#imid_13092067

HPA · ICC-IF U2OSHuman Protein Atlas immunofluorescence, U2OS, antibody CAB036214; green = NME2, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for NME2 IHC & IF

🧪Run kidney first: tubular cells should stain strongly (HPA: High in kidney cells in tubules). Use skeletal muscle myocytes as the negative tissue (HPA: Not detected); on the kidney slide, cells without tubular staining should remain at background, although HPA does not identify a specific internal negative cell population (HPA: kidney cells in tubules).

Positive control tissue: Endometrium (Glandular cells, HPA Medium)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NME2 in U2OS, SK-MEL-30, SiHa, with annotated localisation: Cytosol (supported) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only control; a control matched to the primary antibody’s host species and class (isotype control for a monoclonal or nonimmune IgG for a polyclonal); and, if available, NME2 knockout material or a peptide-blocked primary (standard IHC practice). For chromogenic kidney IHC, quench endogenous peroxidase for HRP detection and check endogenous biotin if using biotin-based detection (standard IHC practice).

⚠️Feasibility: No supplied target or application evidence reports an NME2-specific fixation window or antigen-retrieval effect; the PA1830 intestinal-cancer IHC(P) caption does not report a fixative (catalog caption: PA1830). Optimize retrieval empirically, and assess kidney background from endogenous peroxidase or biotin with the detection controls (standard IHC practice). ICC-IF images show mainly cytosolic NME2 in U2OS, SK-MEL-30 and SiHa, but the supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC (HPA subcellular: cytosol supported).

HPA tissue IHC evidence for NME2

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification. Caution, targets protein from more than one gene.). 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
Endometrium	Glandular cells	Medium	Protein (IHC)	HPA →
Kidney	Cells in tubules	High	Protein (IHC)	HPA →
Lymph node	Non-germinal center cells	High	Protein (IHC)	HPA →
Nasopharynx	Respiratory epithelial cells	High	Protein (IHC)	HPA →
Prostate	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 →
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 NME2 IHC Tips

Use matched tissue controls and compartment-specific scoring to troubleshoot NME2 staining in paraffin sections (HPA tissue IHC; UniProt P22392 localisation).

How should I adjust retrieval when NME2 staining is weak in paraffin sections?

Start with heat-mediated citrate retrieval at pH 6 for the catalog antibody in paraffin sections (datasheet PA1830). Use a consistent heating and cooling cycle across control and study sections, then compare staining in kidney tubules or rectal glandular cells, where high signal is reported (standard IHC practice; HPA: High in kidney tubules and rectal glandular cells). If staining remains weak, test a longer heating interval on matched sections before trying another retrieval buffer as a fallback (standard IHC practice). Judge improvement by cytoplasmic signal relative to background, since general cytoplasmic tissue staining is reported for NME2 (HPA: general cytoplasmic expression).

Could fixation explain weak or uneven NME2 staining?

Target-specific sensitivity of NME2 staining to fixation is unknown from the supplied evidence; the selected tissue image identifies paraffin IHC but does not state its fixative (PA1830 tissue-IHC caption). Record fixative, fixation duration, section thickness and storage conditions for each block, then compare sections processed together using the same retrieval and detection settings (standard IHC practice). If available, use a well-preserved positive control such as kidney tubules beside the problem section to distinguish a run-wide failure from specimen variation (HPA: High in kidney tubules; standard IHC practice). Inspect morphology and staining uniformity before changing antibody conditions, because these checks can reveal processing variation (standard IHC practice).

Should I score nuclear NME2 staining as positive?

Score cytoplasmic staining separately from nuclear staining: tissue IHC reports general cytoplasmic expression, while UniProt also assigns NME2 to the nucleus (HPA: general cytoplasmic expression; UniProt P22392 localisation). A mainly cytosolic signal is supported by subcellular imaging, so an isolated nuclear-only pattern deserves confirmation on a second section or with an independent antibody (HPA: cytosol supported; standard IHC practice). NME2 is also annotated at perinuclear regions, ruffles and lamellipodia during early cell spreading under specified substrate conditions (UniProt P22392 localisation). Keep compartment calls tied to visible cell boundaries and morphology; diffuse chromogen over nuclei alone cannot establish a nuclear pool (standard IHC practice).

How can I assess whether the antibody distinguishes NME2 isoforms and NME1?

The record lists NME2 isoforms 1 and 3, but supplies no mapped epitope or isoform-specific validation for the catalog antibody (UniProt P22392 isoforms; supplied PA1830 evidence). Request its immunogen or epitope information and check which isoform sequences contain that region before assigning isoform identity to tissue staining (standard IHC practice). NME2 can assemble with NME1 in heterohexamers, so complex formation does not itself demonstrate that an NME2 antibody cross-reacts with NME1 (UniProt P22392 subunit). Use an independently validated NME2 reagent or a specific loss-of-target control when the distinction matters, and report unresolved specificity explicitly (standard IHC practice).

How should I examine NME2 by IF alongside a cell-type marker?

For the separate IF/ICC workflow, multiplex NME2 with a validated marker for the cell type under study, then inspect whether signal lies in the same identified cells (standard IF practice). Choose a far-red NME2 fluorophore when the specimen shows strong shorter-wavelength autofluorescence, and include single-color and no-primary controls to assess bleed-through and background (standard IF practice). NME2 has no transmembrane segment and is mainly cytosolic, so permeabilise fixed cells to access its intracellular epitopes; the antibody’s exact epitope is unreported (UniProt P22392 topology; HPA: cytosol supported; supplied PA1830 evidence). Optimise permeabilisation against preserved cell morphology and do not transfer the paraffin IHC caption into an IF fixation claim (standard IF practice; PA1830 tissue-IHC caption).

What should I check when chromogenic NME2 staining appears widespread?

First compare the section with a no-primary control and a matched positive tissue, keeping chromogen development consistent across slides (standard IHC practice). Block endogenous peroxidase before DAB detection, then assess whether diffuse signal persists in the control or accumulates over damaged tissue and section edges (standard chromogenic IHC practice). Widespread cytoplasmic staining can still be plausible because NME2 is ubiquitously expressed and the tissue atlas describes general cytoplasmic expression (UniProt P22392 tissue specificity; HPA: general cytoplasmic expression). If controls indicate background, optimise blocking, washes and antibody concentration incrementally; do not classify every stained cell as specific solely because NME2 is widespread (standard IHC practice; UniProt P22392 tissue specificity).

How should I quantify NME2 across sections with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION

Define the eligible cell population and cytoplasmic compartment before scoring, then use either percent positive cells or an H-score calculated from staining intensity and the fraction of cells at each intensity (HPA: general cytoplasmic expression; standard IHC practice). For spatial comparisons, report positive cells per mm² within a predefined viable tissue region (standard IHC practice). Normalise counts to the number of eligible cells or measured viable area, and keep section processing, imaging and thresholds consistent across groups (standard IHC practice). Record nuclear staining separately, since nuclear localisation is annotated even though the reported tissue IHC pattern is generally cytoplasmic (UniProt P22392 localisation; HPA: general cytoplasmic expression).

How can I distinguish genuine NME2 signal from tissue artefact?

Look for reproducible cellular staining in expected compartments: tissue IHC reports general cytoplasmic expression, with high staining in kidney tubules and rectal glandular cells (HPA: general cytoplasmic expression; HPA: High in kidney tubules and rectal glandular cells). A nuclear component is biologically possible, but a pattern confined to nuclei warrants independent confirmation because the supported subcellular location is mainly cytosolic (UniProt P22392 localisation; HPA: cytosol supported; standard IHC practice). Review cell identity, section edges and necrotic areas against the counterstain, and compare with no-primary and peroxidase-block controls to assess artefact (standard IHC practice). Interpret weak or absent signal cautiously: HPA reports medium agreement with RNA data and warns that its staining may reflect more than one gene (HPA: Approved reliability description).

Boster reagents

Best NME2 / Nucleoside diphosphate kinase B IHC Antibodies

PA1830 has IHC images from human intestinal cancer tissue, rat cerebellum, and mouse intestine, plus IF/ICC images from U20S and HELA cells (PA1830 image captions).

Real IHC data Anti-NME2 antibody, PA1830, IHC(P) IHC(P): Human Intestinal Cancer Tissue

Anti-NME2 Antibody ®

Cat # PA1830

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The rendered SKU is PA1830; its IHC(P) captions show human intestinal cancer tissue, rat cerebellum, and mouse intestine (PA1830 IHC image captions). Its IF/ICC captions show U20S and HELA cells, and the catalog lists Human, Mouse, and Rat reactivity (PA1830 IF/ICC image captions; catalog: PA1830 reactivity).

Which to pick: For tissue IHC, choose PA1830: its own captions document paraffin sections, but do not report the fixative (PA1830 IHC image captions). For IF/ICC, PA1830 has cell image captions and lists both applications (PA1830 IF/ICC image captions; catalog: PA1830 applications). For cross-species IHC, PA1830 has Human, Mouse, and Rat reactivity and IHC captions for each; M01762-1 is listed as a rabbit monoclonal with the same reactivity and IHC/IF applications, but has no supplied IHC/IF image captions (catalog: PA1830 and M01762-1).

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

References

UniProt Consortium. UniProt entry P22392 (NDKB_HUMAN, Nucleoside diphosphate kinase B).
Human Protein Atlas. NME2 tissue IHC expression (reliability: Approved).
Human Protein Atlas. NME2 subcellular location (ICC-IF): Mainly localized to the cytosol..
Human Protein Atlas. NME2 antibody validation summary (5 antibodies).
NME2 reduces proliferation, migration and invasion of gastric cancer cells to limit metastasis. PloS one 2015 — PMC4336288.
NME2 modulates HCC progression through 4EBP1 phosphorylation and autophagy regulation independent of mTOR. Hepatology communications 2025 — PMC12150935.
Non-metastatic 2 (NME2)-mediated suppression of lung cancer metastasis involves transcriptional regulation of key cell adhesion factor vinculin. Nucleic acids research 2014 — PMC4191424.
The therapeutic effects of Bombyx mori sericin on rat skin psoriasis through modulated epidermal immunity and attenuated cell proliferation. Journal of traditional and complementary medicine 2021 — PMC8572706.
PubMed PMID:1988104 — UniProt-cited evidence.
PubMed PMID:1851158 — UniProt-cited evidence.
PubMed PMID:8392752 — UniProt-cited evidence.