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- Table of Contents
Plan paraffin-section MTA2 IHC around the widespread nuclear staining observed in tissue (HPA tissue IHC). This guide covers antibody dilution, chromogenic detection and nuclear scoring, using 0.5–1 μg/mL as the catalog antibody A03073-1 starting range (datasheet: A03073-1).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Nuclear staining across tissues (HPA tissue IHC) | |
| Staining pattern | Widespread nuclear staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03073-1) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Resident cells may add nuclear signal in mixed tissues (HPA tissue IHC) | |
| Regulation | Widely expressed (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences are unspecified (UniProt) |
The catalog antibody’s IHC-P protocol (datasheet: A03073-1) is accompanied by 3 published MTA2 IHC protocols (PMC6393561; PMC10296612; PMC4634783).
| Sample | Paraffin-embedded human glioma tissue; fixative not specified (datasheet A03073-1) |
| Fixation | Image fixative and duration unreported (datasheet A03073-1); 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 A03073-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03073-1) |
| Primary antibody | Rabbit anti-MTA2, 0.5-1μg/ml (datasheet A03073-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03073-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03073-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MTA2-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
MTA2 is a nuclear, non-transmembrane protein (UniProt O94776: subcellular location and topology). In paraffin-section IHC, expect nuclear staining across many cell types: HPA describes ubiquitous nuclear expression and reports high staining in several named populations (HPA: tissue IHC). The tissue pattern is rated Supported because antibody staining is highly consistent with RNA expression; that rating does not establish how fixation affects this target (HPA: tissue IHC reliability).
| Distinct chromogenic staining in nuclei of glandular cells in breast, appendix or adrenal gland. | This matches the reported compartment and high-staining cell populations (HPA: tissue IHC). Judge nuclear contrast against adjacent background; high staining in these examples does not imply that every cell on the section must have identical intensity (HPA: tissue IHC; general IHC interpretation). |
| Predominantly cytoplasmic, surface-like or extracellular staining with little nuclear signal. | Treat this as a discordant pattern requiring validation: UniProt places MTA2 in the nucleus and reports no transmembrane segment, while HPA describes nuclear tissue staining (UniProt O94776: location and topology; HPA: tissue IHC). Distribution alone cannot identify the artefact's cause (general IHC interpretation). |
| The strongest signal appears in cells outside the reported high-staining population of the sampled tissue. | Check cell identity and assay controls before calling it MTA2: cross-reactivity or endogenous chromogen-generating activity can mimic a positive result (general IHC practice). HPA lists selected high-staining populations, but supplies no negative-cell list here, so staining elsewhere is not automatically false (HPA: tissue IHC). |
| A diffuse haze covers nuclei, cytoplasm and tissue spaces, obscuring cell boundaries. | This is poor compartment resolution rather than convincing nuclear localisation (UniProt O94776: nuclear location; general IHC interpretation). Review background in a reagent control and compare the staining and wash conditions with a clean run (general IHC practice). |
| No nuclear signal appears in an otherwise interpretable section of a reported high-staining tissue. | A failed or insensitive assay is plausible, especially when an appropriate control also lacks signal (HPA: high staining in the listed tissues; general IHC practice). A single negative section cannot establish absence of MTA2, since the HPA tissue profile is broad and is not a guarantee for every specimen (HPA: tissue IHC). |
| Compartment and distribution | MTA2 is nuclear in UniProt, and HPA describes ubiquitous nuclear tissue expression (UniProt O94776: location; HPA: tissue IHC). Score localisation and the stained cell population separately; a dark chromogen deposit outside nuclei does not establish target-specific expression (general IHC interpretation). |
| Choice of comparison tissue | HPA reports high staining in adipocytes, bone-marrow hematopoietic cells, bronchial respiratory epithelial cells and caudate glial cells, among others (HPA: tissue IHC). These are documented comparison populations, not a ranking of assay performance or a promise that all specimens stain equally (HPA: tissue IHC; general IHC interpretation). |
| Antibody evidence | HPA lists IHC as Supported for HPA006214, CAB005315 and CAB017522; no IHC status is supplied for HPA072727 (HPA: antibody validation). That evidence supports a reported staining pattern for those antibodies, but does not validate an unspecified catalog antibody or its dilution (HPA: antibody validation). |
| Isoforms and processing | UniProt records two isoforms and a full-length chain of residues 1–668, with no signal peptide or propeptide (UniProt O94776: isoforms and processing). Antibody epitope information is needed to judge isoform coverage; the record alone does not predict differing IHC patterns or shedding (UniProt O94776: isoforms and processing). |
| Detection chemistry | Endogenous enzyme activity can create chromogenic signal, so interpret suspected activity with a suitable reagent control and the assay's blocking steps (general IHC practice). The supplied HPA pattern does not identify endogenous activity in any particular tissue as an MTA2-specific problem (HPA: tissue IHC). |
| IF/ICC cross-check | Q: Where should MTA2 appear by IF/ICC? A: HPA reports enhanced nucleoplasmic localisation; use that as a compartment cross-check, while interpreting the paraffin-section chromogenic result on its own evidence (HPA: subcellular ICC-IF; HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Only cytoplasmic or membranous colour is visible. | The compartment conflicts with nuclear MTA2 and its lack of a transmembrane segment; the precise cause remains unresolved (UniProt O94776: location and topology). | Confirm nuclear counterstain and tissue morphology, then compare a documented high-staining population and a reagent control (HPA: tissue IHC; general IHC practice). |
| High colour appears in cells that were not expected to dominate the sampled tissue. | Misidentified cells, cross-reactivity or endogenous detection activity are possibilities (general IHC practice); HPA provides no negative-cell list for this record (HPA: tissue IHC). | Recheck cell identity on the counterstained section and inspect detection controls before assigning cell-specific MTA2 positivity (general IHC practice). |
| Brown signal is diffuse and nuclear borders are hard to resolve. | Background deposition or excessive staining can obscure the expected nuclear pattern (UniProt O94776: location; general IHC practice). | Compare a reagent control; review antibody dilution, detection time, blocking and washes using the chosen assay instructions (general IHC practice). No MTA2-specific dilution is supplied. |
| A reported high-staining comparison tissue has no nuclear signal. | An assay or detection failure is possible; HPA's high category does not guarantee a positive result in every specimen (HPA: tissue IHC; general IHC interpretation). | Check reagent controls and the validated assay instructions, including antigen retrieval and detection steps (general IHC practice). Target-specific fixation sensitivity is unreported in the supplied sources. |
| Different antibodies produce conflicting nuclear patterns. | The supplied HPA entries have different IHC evidence: three are Supported, while one has no IHC status listed (HPA: antibody validation). Epitope and isoform coverage cannot be inferred here (UniProt O94776: isoforms). | Compare each antibody's documented IHC status and available epitope information, then repeat against the same tissue and controls (HPA: antibody validation; general IHC practice). |
| IF/ICC looks nucleoplasmic, but IHC shows widespread non-nuclear colour. | The IF compartment agrees with HPA's nucleoplasmic observation; the IHC compartment conflicts with the reported nuclear tissue pattern (HPA: subcellular ICC-IF; HPA: tissue IHC). | Evaluate the IHC section's background and detection controls, then score its nuclear staining independently; an IF image does not by itself validate that chromogenic result (general IHC interpretation). |
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — High 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.
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: MTA2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Troubleshoot MTA2 staining in paraffin sections by checking retrieval, nuclear localisation, background and scoring against the reported IHC conditions and tissue profile.
IHC images cover human tissue (catalog IHC captions), and IF/ICC images cover human cells (catalog IF captions). A03073 and A03073-1 also list mouse and rat reactivity (catalog reactivity).
A03073-1 shows IHC in human glioma paraffin sections and IF/ICC in A549 cells (A03073-1 image captions); M03073 shows IHC in human cervical and stomach cancer paraffin sections (M03073 IHC captions). A03073 shows IHC in human small intestine and IF/ICC in HeLa cells (A03073 image captions).
Which to pick: For tissue IHC, choose A03073-1 for human glioma paraffin sections or the rabbit monoclonal M03073 for human cervical cancer paraffin sections (A03073-1 and M03073 IHC captions; M03073 catalog clone). For IF/ICC, A03073-1 has an A549 cell image and A03073 has HeLa cell images (catalog IF captions). For mouse or rat work, consider A03073-1 or A03073 because both list those species and IHC and IF applications (catalog reactivity and applications); their mouse and rat performance is not shown in the supplied images (catalog image captions). The fixative is unreported in the tissue IHC captions (catalog IHC captions).