MAML2 / Mastermind-like protein 2 · IHC design guide

Design Immunohistochemistry for MAML2

Plan MAML2 paraffin-section IHC with a 1:25 starting dilution for the catalog antibody (datasheet M03679-1). Assess nuclear and cytoplasmic staining in tissue (HPA tissue IHC), while treating punctate nuclear localisation as a molecular expectation (UniProt).

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

Immunohistochemistry protocol sheet for MAML2 (IHC for MAML2): expected localisation Nuclear and cytoplasmic (HPA tissue IHC); punctate nuclear location (UniProt), antibody M03679-1, validated IHC image, and IHC protocol steps — Printable **MAML2** IHC protocol sheet — expected localisation Nuclear and cytoplasmic (HPA tissue IHC); punctate nuclear location (UniProt), antibody M03679-1, controls and protocol steps. Open the full MAML2 IHC guide →

Navigate to: Protocols Staining pattern Controls Antibodies

MAML2 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	Nuclear and cytoplasmic (HPA tissue IHC); punctate nuclear location (UniProt)
	Staining pattern	General nuclear and cytoplasmic staining (HPA tissue IHC)
	Antigen retrieval	Citrate pH 6 HIER, heat-mediated (datasheet M03679-1)
	Positive control ⓘ	Adrenal gland+4 more · see all
	Negative control ⓘ	Parathyroid gland

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep fixation consistent across sections (standard IHC practice; not target-specific)
	Caveat	Medium agreement between staining and RNA expression (HPA tissue IHC)
	Regulation	Staining-intensity regulation unreported (UniProt)
	Isoform / epitope	No isoforms annotated; one 1–1156 chain (UniProt)

Section 1

Recommended MAML2 IHC & IF Protocols

The catalog antibody’s IHC-P protocol uses citrate retrieval (datasheet M03679-1). One published MAML2 IHC study provides a sinonasal papilloma tissue microarray protocol (PMC10907326).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Formaldehyde-fixed, paraffin-embedded human kidney tissue (datasheet M03679-1)
Fixation	Image formalin-fixed; duration unreported (datasheet M03679-1); verify before use.
Sectioning	4–5 µm sections on charged slides (standard)
Deparaffinisation	Xylene, graded ethanol series to water (standard)
Antigen retrieval	Heat retrieval: Citrate pH 6 (datasheet M03679-1); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% normal serum of the secondary host, 30 min, room temperature (standard)
Primary antibody	Rabbit anti-MAML2, 1:25 (datasheet M03679-1)
Primary incubation	1 hours at 37°C (datasheet M03679-1)
Detection	HRP-polymer secondary, DAB chromogen 5–10 min (standard)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	MAML2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General nuclear and cytoplasmic expression. No signal in the no-primary control.

💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet M03679-1); the published study does not specify its antigen-repair conditions (PMC10907326).

Section 2

What Is the Expected MAML2 Staining Pattern?

MAML2 is a nuclear protein with a punctate pattern in nuclear speckles (UniProt Q8IZL2). HPA describes general nuclear and cytoplasmic tissue staining, with high signal in several epithelial and glandular cell types (HPA tissue IHC). Its tissue IHC reliability is Approved, with medium consistency between antibody staining and RNA expression (HPA tissue IHC). MAML2 has no transmembrane segment (UniProt Q8IZL2 topology).

What am I looking at on my slide?
Nuclear staining in adrenal, colon or duodenal glandular cells.	This fits the reported high staining in these cell types (HPA tissue IHC). Assess the signal within the relevant cells; staining elsewhere in the section does not establish the expected pattern (general IHC practice).
Punctate nuclear staining, with some cytoplasmic signal.	Punctate nuclear localisation is consistent with nuclear speckles (UniProt Q8IZL2) and supported nuclear bodies in ICC-IF (HPA subcellular). HPA also reports general cytoplasmic tissue staining, so cytoplasmic signal alone is not automatically an artefact (HPA tissue IHC).
Predominantly membranous or extracellular staining, without convincing nuclear signal.	This does not match the reported nuclear localisation (UniProt Q8IZL2; HPA subcellular). Because MAML2 has no transmembrane segment, review such staining for nonspecific binding or detection artefact before assigning it to MAML2 (UniProt Q8IZL2 topology; general IHC practice).
Strong staining in parathyroid glandular cells.	HPA reports MAML2 as not detected in these cells (HPA tissue IHC). Check for cross-reactivity or endogenous detection activity; a single discordant section does not establish which explanation applies (general IHC practice).
Diffuse colour across cells, stroma and blank areas.	This distribution cannot be scored as cell-specific MAML2 staining (general IHC practice). Compare with a no-primary control and inspect blocking, washing and detection steps for background (general IHC practice).

💡Expected MAML2 appearanceCall a result positive when relevant cells show interpretable nuclear staining, potentially punctate, at a level consistent with the tissue reference; high glandular-cell staining in colon is one example (UniProt Q8IZL2; HPA tissue IHC). Diffuse colour outside cell boundaries is background, not a positive cell score (general IHC practice).

How each factor affects the staining
Tissue and cell selection	HPA reports high staining in adrenal, colon and duodenal glandular cells, bronchial respiratory epithelium, cerebellar Purkinje cells, esophageal squamous epithelium and kidney glomerular cells (HPA tissue IHC). Parathyroid glandular cells are reported as not detected (HPA tissue IHC).
Compartment and imaging method	UniProt describes punctate nuclear localisation; HPA tissue IHC describes general nuclear and cytoplasmic staining (UniProt Q8IZL2; HPA tissue IHC). Supported nuclear bodies and nucleoplasm come from ICC-IF, so do not require individual nuclear puncta to be resolved in every chromogenic section (HPA subcellular; general IHC practice).
Strength of antibody evidence	The HPA tissue IHC profile is Approved with medium staining-to-RNA consistency, and HPA035223 is IHC Approved (HPA tissue IHC; HPA antibodies). Its ICC validation is Uncertain, which limits what that antibody alone can establish about IF localisation (HPA antibodies).
Protein topology and processing	MAML2 has no transmembrane segment or annotated signal peptide and is recorded as a single 1–1156 chain (UniProt Q8IZL2). These annotations support scrutinising a membrane-dominant result; they do not establish an antigen-retrieval setting or a fixation effect (UniProt Q8IZL2; general IHC practice).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
No staining in a chosen positive tissue.	The reference identifies high staining only in specified cell types; sampling the wrong cells can make a section appear negative (HPA tissue IHC). A failed staining run is another possibility (general IHC practice).	Confirm the expected cells are present, then review the run's positive control, primary-antibody step and detection reagents (HPA tissue IHC; general IHC practice).
Only broad cytoplasmic colour is visible.	HPA allows cytoplasmic tissue staining, but a result lacking interpretable nuclear staining is less consistent with UniProt nuclear localisation (HPA tissue IHC; UniProt Q8IZL2).	Compare cell boundaries and nuclei with the counterstain and a no-primary control; score only staining that can be assigned to cells (general IHC practice).
Membrane edges or extracellular material dominate.	This pattern conflicts with reported nuclear localisation and the absence of a transmembrane segment (UniProt Q8IZL2). Its cause cannot be identified from appearance alone (general IHC practice).	Check the no-primary control, blocking and detection background before interpreting the signal (general IHC practice).
Parathyroid glandular cells appear strongly positive.	This conflicts with HPA's not-detected reference for those cells; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice).	Verify cell identity, compare a no-primary control and review the detection system; report the disagreement rather than treating this tissue as a confirmed positive control (general IHC practice).
Background obscures cell-level scoring.	Diffuse staining can arise from nonspecific binding or detection background (general IHC practice); HPA's tissue profile cannot identify the cause in this run (HPA tissue IHC).	Inspect no-primary controls and optimise blocking, washing or detection conditions according to the validated IHC workflow (general IHC practice).
Can IF/ICC confirm the punctate nuclear location?	HPA supports nuclear bodies and nucleoplasm in ICC-IF, but rates HPA035223 ICC validation Uncertain (HPA subcellular; HPA antibodies).	Use the separate IF/ICC guide to plan that experiment; interpret its images alongside the validation status rather than applying an IF protocol from this IHC section (HPA antibodies; general IHC practice).

Source: UniProt Q8IZL2 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing MAML2 staining

M03679-1 staining MAML2 in human kidney tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.

M03679-1 · IHCM03679-1 staining MAML2 in human kidney tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections).

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000184384-MAML2/tissue/cerebral+cortex#imid_10180001

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000184384-MAML2/tissue/colon#imid_10188639

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000184384-MAML2/tissue/liver#imid_10188648

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000184384-MAML2/tissue/kidney#imid_10188649

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035223; green = MAML2, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for MAML2 IHC & IF

🧪Run adrenal gland first and require nuclear staining in its glandular cells (HPA: High in adrenal gland glandular cells; UniProt Q8IZL2: punctate nuclear localization). Use parathyroid glandular cells as the negative tissue (HPA: Not detected in parathyroid gland glandular cells); on the adrenal slide, cells outside the glandular compartment should lack comparable specific nuclear staining, but should not be treated as a validated negative population.

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

Negative control tissue: Parathyroid gland (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MAML2 in A-431, U-251MG, U2OS, with annotated localisation: Nuclear bodies (supported) (HPA subcellular).

Technical controls: Include a no-primary (secondary-only) slide, an isotype control matched to the primary antibody’s host species, immunoglobulin class and clonality, and a MAML2 knockout specimen or validated peptide-block control. Quench endogenous peroxidase and block endogenous biotin where needed, then check the no-primary slide for background from the biotinylated secondary (selected-SKU tissue-IHC caption: biotinylated secondary).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU tissue-IHC caption lists the fixative as unreported (selected-SKU tissue-IHC caption: fixative not stated). The caption uses heat-mediated citrate retrieval at pH 6, but does not establish whether retrieval is required; the supplied IHC-P evidence also does not establish that frozen sections or IF are easier (selected-SKU tissue-IHC caption: citrate retrieval). No adrenal-specific artefact is established by the supplied evidence; assess chromogenic background with the no-primary control.

HPA tissue IHC evidence for MAML2

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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
Adrenal gland	Glandular cells	High	Protein (IHC)	HPA →
Bronchus	Respiratory epithelial cells	High	Protein (IHC)	HPA →
Cerebellum	Purkinje cells	High	Protein (IHC)	HPA →
Colon	Glandular cells	High	Protein (IHC)	HPA →
Duodenum	Glandular cells	High	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Parathyroid gland	Glandular cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced MAML2 IHC Tips

Troubleshoot MAML2 chromogenic IHC in paraffin sections by checking retrieval, staining compartment, controls and scoring before interpreting signal.

What retrieval should I use when MAML2 staining is weak in paraffin sections?

Use heat-mediated antigen retrieval in citrate buffer at pH 6 (datasheet M03679-1). The selected antibody’s kidney IHC example used that retrieval with primary antibody at 1:25 for 1 hour at 37°C (caption M03679-1). If staining remains weak, adjust heating duration in small steps while holding section preparation, antibody concentration and detection constant (standard IHC practice). Compare each condition against a positive control and a section processed without primary antibody; excess heating can damage tissue morphology or increase nonspecific staining (standard IHC practice). Judge improvement by interpretable nuclear staining, including puncta, rather than total chromogen intensity alone (UniProt Q8IZL2 localisation; standard IHC practice).

How should I troubleshoot weak MAML2 staining after fixation?

Target-specific fixation sensitivity for MAML2 is unknown from the supplied evidence; do not assign weak staining to a particular fixative without a controlled comparison. The selected kidney image documents formaldehyde-fixed paraffin tissue, but gives no comparison of fixatives or fixation durations (caption M03679-1). Record the fixative, exposure time, section age and processing conditions for each case, then compare matched sections using the same citrate pH 6 retrieval and detection workflow (datasheet M03679-1; standard IHC practice). If nuclear detail is poor throughout a section, assess tissue preservation and retrieval before changing antibody concentration (standard IHC practice). Include a consistently processed positive control in each run (standard IHC practice).

Should MAML2 appear exclusively in nuclei?

Expect a punctate nuclear pattern because MAML2 is annotated in nuclear speckles (UniProt Q8IZL2 localisation). Nuclear bodies are its supported main location, with supported nucleoplasmic and approved cytosolic localisation in cell imaging (HPA subcellular). Tissue IHC is described more broadly as nuclear and cytoplasmic, so cytoplasmic colour alone cannot establish either specificity or artefact (HPA tissue IHC). Assess whether discrete nuclear signal survives comparison with the no-primary control, and score nuclear and cytoplasmic compartments separately (standard IHC practice). Diffuse staining across every compartment, especially where tissue morphology is damaged, warrants a review of blocking, detection and counterstain (standard IHC practice).

Could epitope placement change how I interpret MAML2 staining?

The supplied record annotates one MAML2 chain spanning residues 1–1156, with no annotated isoforms or transmembrane segment (UniProt Q8IZL2 record). Its N-terminal region interacts with NOTCH proteins, and a phosphoserine is annotated at residue 175 (UniProt Q8IZL2 record). Those annotations do not identify the catalog antibody’s epitope or show that binding depends on phosphorylation (supplied record; datasheet M03679-1). Check the antibody’s documented immunogen or epitope before assigning staining differences to an isoform or modification (standard IHC practice). If epitope information is unavailable, compare staining patterns with an independently validated antibody and appropriate controls before making that assignment (standard IHC practice).

How can I cross-check MAML2 localisation by IF alongside tissue IHC?

Use IF as a separate localisation check and multiplex MAML2 with a validated marker for the cell type under study (standard IF practice). Choose fluorophores and imaging channels after measuring tissue autofluorescence; reserve the cleaner channel for a weak MAML2 signal (standard IF practice). MAML2 has no annotated transmembrane segment and is principally associated with nuclear bodies, so permit access to intracellular epitopes with controlled permeabilisation (UniProt Q8IZL2 topology; HPA subcellular; standard IF practice). Its antibody epitope is unspecified, so optimise permeabilisation empirically and include single-colour and no-primary controls (datasheet M03679-1; standard IF practice). Compare nuclear puncta with the chromogenic section without assuming identical signal intensity across methods (UniProt Q8IZL2 localisation; standard imaging practice).

What should I check if chromogenic MAML2 staining covers the whole section?

First compare the section with a no-primary control to locate background from secondary detection or endogenous enzyme activity (standard IHC practice). The selected kidney example used 3% BSA for 0.5 hour and an undiluted biotinylated secondary antibody, so assess blocking and detection controls when following that workflow (caption M03679-1). For a peroxidase and DAB workflow, include a peroxidase block and check endogenous biotin where biotin-based detection is used (standard IHC practice). Titrate primary antibody around the example’s 1:25 condition while holding retrieval and detection constant (caption M03679-1; standard IHC practice). Read questionable signal alongside intact nuclear morphology and the expected punctate pattern (UniProt Q8IZL2 localisation; standard IHC practice).

How should I score MAML2 IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION

Define the analysis compartment and count eligible, intact cells before comparing cases (standard IHC scoring practice). Report the percentage of positive nuclei and an H-score from 0–300, calculated from the percentages of cells at intensity levels 0–3; report cytoplasmic staining separately (standard IHC scoring practice; HPA tissue IHC). If the question concerns spatial abundance, report positive cells per mm² of viable tissue as an additional measure (standard IHC scoring practice). Normalise counts to the relevant cell population or viable tissue area, with the same staining threshold and acquisition conditions throughout (standard IHC scoring practice). Set thresholds using positive and no-primary controls, then apply them blinded to sample identity (standard IHC scoring practice).

How do I distinguish convincing MAML2 signal from staining artefact?

Give most weight to reproducible nuclear signal with puncta, consistent with MAML2 nuclear speckles and supported nuclear-body localisation (UniProt Q8IZL2 localisation; HPA subcellular). Check cellular context: HPA reports high staining in kidney glomerular cells and no detection in parathyroid glandular cells, while its tissue IHC has medium consistency with RNA expression (HPA tissue IHC). Treat staining confined to section edges, necrotic regions or damaged cells cautiously, and compare it with adjacent intact tissue (standard IHC practice). Use no-primary and detection controls to identify nonspecific colour or endogenous enzyme signal (standard IHC practice). Cytoplasmic staining requires its own assessment because HPA also reports cytoplasmic tissue expression (HPA tissue IHC).

Boster reagents

Best MAML2 / Mastermind-like protein 2 IHC Antibodies

Anti-MAML2 options cover human and rat IHC; the supplied figure shows human kidney paraffin sections, with no IF/ICC data provided (catalog applications and reactivity; M03679-1 image caption).

Real IHC data

Anti-MAML2 Antibody (Center)

Cat # M03679-1

View product →

M03679-1 is listed for human IHC-P and has an IHC figure from formaldehyde-fixed human kidney paraffin sections (M03679-1 catalog applications; image caption). A03679-1 lists IHC for human and rat, but has no supplied IHC figure or reported fixative (A03679-1 catalog applications, reactivity and image fields).

Which to pick: For human tissue IHC-P, choose M03679-1: it is a rabbit polyclonal antibody with a documented 1:25 dilution and a human kidney paraffin-section figure (M03679-1 catalog host and dilution; image caption). For human and rat IHC, A03679-1 is the rabbit polyclonal option listed for both species, although its processing and fixative are unreported (A03679-1 catalog applications, reactivity, host and image fields). Neither SKU lists IF/ICC validation or an IF dilution, so neither has payload support as an IF/ICC choice (catalog applications and IF dilution fields).

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

References

UniProt Consortium. UniProt entry Q8IZL2 (MAML2_HUMAN, Mastermind-like protein 2).
Human Protein Atlas. MAML2 tissue IHC expression (reliability: Approved).
Human Protein Atlas. MAML2 subcellular location (ICC-IF): Mainly localized to the nuclear bodies. In addition localized to the nucleoplasm and cytosol..
Human Protein Atlas. MAML2 antibody validation summary (1 antibodies).
Comprehensive analysis of DNA methylation gene expression profiles in GEO dataset reveals biomarkers related to malignant transformation of sinonasal inverted papilloma. Discover oncology 2024 — PMC10907326.
MAML2 rearrangement in primary pulmonary mucoepidermoid carcinoma and the correlation with FLT1 expression. PloS one 2014 — PMC3979848.
Malignant undifferentiated epithelioid neoplasms with MAML2 rearrangements: A clinicopathologic study of seven cases demonstrating a heterogenous entity. Genes, chromosomes & cancer 2023 — PMC9908836.
A reappraisal of the MECT1/MAML2 translocation in salivary mucoepidermoid carcinomas. The American journal of surgical pathology 2010 — PMC13401325.
PubMed PMID:12539049 — UniProt-cited evidence.
PubMed PMID:11347906 — UniProt-cited evidence.
PubMed PMID:16554811 — UniProt-cited evidence.