MTA2 / Metastasis-associated protein MTA2 · IHC design guide

Design Immunohistochemistry for MTA2

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).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for MTA2 (IHC for MTA2): expected localisation Nuclear staining across tissues (HPA tissue IHC), antibody A03073-1, validated IHC image, and IHC protocol steps
Printable MTA2 IHC protocol sheet — expected localisation Nuclear staining across tissues (HPA tissue IHC), antibody A03073-1, controls and protocol steps. Open the full MTA2 IHC guide →

MTA2 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 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
Important caveats
Reasons your staining may differ from the expected pattern.
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)
Section 1

Recommended MTA2 IHC & IF Protocols

The catalog antibody’s IHC-P protocol (datasheet: A03073-1) is accompanied by 3 published MTA2 IHC protocols (PMC6393561; PMC10296612; PMC4634783).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human glioma tissue; fixative not specified (datasheet A03073-1)
FixationImage fixative and duration unreported (datasheet A03073-1); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: EDTA pH 8.0 (datasheet A03073-1); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet A03073-1)
Primary antibodyRabbit anti-MTA2, 0.5-1μg/ml (datasheet A03073-1)
Primary incubationOvernight at 4 °C (datasheet A03073-1)
DetectionStreptavidin-biotin complex (SABC), DAB chromogen (datasheet A03073-1)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultMTA2-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control.
💡Decision noteStart with heat-mediated EDTA at pH 8.0 for the catalog antibody (datasheet: A03073-1); the published citrate protocols used pH 6.0 (PMC6393561; PMC10296612).
Section 2

What Is the Expected MTA2 Staining Pattern?

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).

What am I looking at on my slide?
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).
💡Expected MTA2 appearanceCall a result consistent with MTA2 when nuclei are clearly stained in the relevant cells, including reported high-staining populations; dominant non-nuclear colour is a discordant pattern to investigate (HPA: tissue IHC; UniProt O94776: nuclear location; general IHC interpretation).
How each factor affects the staining
Compartment and distributionMTA2 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 tissueHPA 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 evidenceHPA 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 processingUniProt 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 chemistryEndogenous 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-checkQ: 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).
Why is my staining missing, weak or wrong?
SituationLikely causeNext 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).

Sample controls for MTA2 IHC & IF

🧪Run breast tissue first and assess glandular cells for nuclear staining (HPA: High in breast glandular cells; UniProt O94776: nucleus). HPA lists no negative tissue and detects MTA2 in all 45 scored tissues, so no-primary and isotype controls carry the negative comparison; no cell type on the breast slide is established as an internal negative, and cells without signal should show counterstain without nuclear chromogen (HPA: no negative tissue listed).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: MTA2 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 MTA2 in A-431, U-251MG, U2OS, HAP1, REH, U2OS, siRNA 1 (10x), U2OS, siRNA 2 (10x), U2OS, scrambled (10x), with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide, a concentration-matched rabbit IgG isotype control matched to the primary’s clonality where known, and a processed MTA2-knockout sample as a biological negative (caption: rabbit primary antibody; standard IHC controls). Quench endogenous peroxidase and check endogenous biotin background in breast sections before interpreting DAB staining (caption: biotinylated secondary, streptavidin–biotin detection and DAB; standard IHC practice).
⚠️Feasibility: The selected paraffin-section example uses heat retrieval in EDTA at pH 8.0, but reports no fixative; a target-specific fixation window or fixation effect is unreported (caption: EDTA retrieval; fixative not stated). Paraffin IHC is directly demonstrated, while HPA ICC-IF images support nucleoplasmic localization without establishing that IF or frozen sections are easier (caption: paraffin-section IHC; HPA: nucleoplasm, enhanced). In breast sections, glandular and surrounding tissue background should be checked against the negative controls, particularly with biotin-based detection (caption: streptavidin–biotin detection; standard IHC practice).

HPA tissue IHC evidence for MTA2

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.

Positive expression · recommended positive controls

TissueCell typeLevelEvidenceSource
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 →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
None in HPA: MTA2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
Section 3

Advanced MTA2 IHC Tips

Troubleshoot MTA2 staining in paraffin sections by checking retrieval, nuclear localisation, background and scoring against the reported IHC conditions and tissue profile.

How should I retrieve MTA2 antigen when nuclear staining is weak?
Start with heat-mediated retrieval in EDTA, pH 8.0, the condition reported for the catalog antibody in a paraffin section (datasheet A03073-1). The reported human glioma workflow then used 10% goat serum and 1 µg/ml primary antibody overnight at 4°C (caption A03073-1). If nuclei remain weak, compare retrieval heating and cooling across matched sections while holding antibody concentration and detection constant (standard IHC practice). Assess staining in intact nuclei against a matched control section before increasing retrieval intensity, since tissue damage can make apparent signal difficult to interpret (standard IHC practice; UniProt O94776: nucleus).
Can fixation explain weak or uneven MTA2 staining?
Target-specific sensitivity of MTA2 to fixation is unknown from the supplied evidence; the selected tissue caption identifies a paraffin section but does not state its fixative (caption A03073-1). Record the actual fixative, fixation interval and tissue thickness for each specimen before comparing nuclear signal across cases (standard IHC practice). Keep processing and EDTA, pH 8.0 retrieval consistent across comparison sections, then inspect preservation alongside chromogenic staining (datasheet A03073-1; standard IHC practice). If signal varies with processing, test matched sections under controlled conditions and report the observed effect as experimental, without assigning MTA2-specific fixation sensitivity (standard IHC practice).
Where should convincing MTA2 staining appear in a tissue section?
Evaluate MTA2 primarily within nuclei: UniProt assigns it to the nucleus, and HPA reports ubiquitous nuclear tissue staining and enhanced nucleoplasmic localisation (UniProt O94776; HPA tissue IHC; HPA subcellular). In the selected paraffin-section example, the antibody was applied to human glioma tissue at 1 µg/ml overnight at 4°C (caption A03073-1). Compare nuclear signal with the counterstain in intact cells, checking whether chromogen occupies the expected compartment rather than surrounding cytoplasm (standard IHC practice; UniProt O94776: nucleus). Broad staining across cell populations can fit the reported ubiquitous profile, so identify cells morphologically before interpreting differences in intensity (HPA tissue IHC; standard IHC practice).
Could isoforms or epitope accessibility change my MTA2 IHC result?
MTA2 has 2 listed isoforms, but the supplied antibody caption does not identify its epitope or establish isoform recognition (UniProt O94776; caption A03073-1). Its annotated BAH (1–144), ELM2 (145–256) and SANT (263–315) regions provide sequence context, not evidence that this antibody binds any one region (UniProt O94776). Annotated phosphorylation and acetylation sites could matter to an epitope only if binding and site overlap were demonstrated; that overlap is unreported here (UniProt O94776; caption A03073-1). Compare staining under the reported EDTA, pH 8.0 retrieval condition and document antibody identity before attributing a difference to isoforms or modifications (datasheet A03073-1; standard IHC practice).
How can IF help investigate an ambiguous MTA2 IHC pattern?
Use IF as a complementary localisation check: HPA reports enhanced nucleoplasmic MTA2 signal in its subcellular data, while the selected antibody caption documents chromogenic tissue IHC (HPA subcellular; caption A03073-1). For multiplexing, pair MTA2 with a marker of the cell population under study, such as a glial cell marker when examining the HPA-reported glial population in caudate (HPA tissue IHC; standard IF practice). Choose a fluorophore channel with low measured tissue autofluorescence and include single-channel controls before assigning nuclear overlap (standard IF practice). Because MTA2 is nuclear and has no transmembrane segment, permeabilise sufficiently for nuclear access, then assess signal within segmented nuclei; optimise IF conditions independently of the IHC caption (UniProt O94776; standard IF practice).
What should I check when DAB background obscures MTA2 nuclei?
The reported IHC example used 10% goat serum, a biotinylated secondary antibody, streptavidin–biotin detection and DAB chromogen (caption A03073-1). Check a section without primary antibody to separate detection-system staining from primary-dependent nuclear signal, and inspect whether background follows tissue structures rather than nuclei (standard IHC practice; UniProt O94776: nucleus). Apply an appropriate endogenous peroxidase block and assess background from the biotin-based detection workflow as general chromogenic IHC controls (standard IHC practice; caption A03073-1). If staining remains diffuse, titrate the primary around the reported 1 µg/ml condition while keeping retrieval and development comparable between sections (caption A03073-1; standard IHC practice).
How should I score MTA2 across heterogeneous tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Score MTA2 in morphologically intact nuclei, since its annotated location and the HPA tissue profile are nuclear (UniProt O94776; HPA tissue IHC). Record the percentage of positive nuclei and an intensity-based H-score for each defined cell population, using consistent thresholds across the batch (standard IHC practice). Normalise positive counts to all eligible nuclei in that population, or report positive nuclei per mm² of evaluable tissue when density is the outcome (standard IHC practice). Exclude necrotic or poorly preserved regions, document the sampled area, and compare sections stained with the same EDTA, pH 8.0 retrieval and detection settings (standard IHC practice; datasheet A03073-1).
How do I distinguish genuine MTA2 positivity from staining artefact?
A convincing result places chromogen in intact nuclei, consistent with UniProt nuclear annotation and HPA's ubiquitous nuclear tissue profile (UniProt O94776; HPA tissue IHC). Unexpected cytoplasmic deposits or staining confined to an implausible cell population deserve review against morphology and controls; HPA lists high staining in several distinct cell populations (HPA tissue IHC; standard IHC practice). Discount signal concentrated at section edges or necrotic areas until it reproduces in well-preserved interior tissue (standard IHC practice). Because the reported workflow uses biotin-based detection and DAB, compare a no-primary control and assess endogenous enzyme background before interpreting faint deposits as MTA2 expression (caption A03073-1; standard IHC practice).
Boster reagents

Best MTA2 / Metastasis-associated protein MTA2 IHC Antibodies

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).

Real IHC data IHC analysis of MTA2 using anti-MTA2 antibody (A03073-1). MTA2 was detected in paraffin-embedded section of human glioma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-MTA2 Antibody (A03073-1) 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.
Anti-MTA2/PID ® Antibody
Cat # A03073-1
Real IHC data IHC analysis of MTA2 using anti-MTA2 antibody (M03073). MTA2 was detected in a paraffin-embedded section of human cervical cancer 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 1:50 rabbit anti-MTA2 Antibody (M03073) 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.
Anti-MTA2 Monoclonal Antibody
Cat # M03073
Real IHC data Immunohistochemistry of PID in human small intestine tissue with PID antibody at 2.5 μg/ml.
Anti-PID MTA2 Antibody
Cat # A03073

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).

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

References

  1. UniProt Consortium. UniProt entry O94776 (MTA2_HUMAN, Metastasis-associated protein MTA2).
  2. Human Protein Atlas. MTA2 tissue IHC expression (reliability: Supported).
  3. Human Protein Atlas. MTA2 subcellular location (ICC-IF): Localized to the nucleoplasm..
  4. Human Protein Atlas. MTA2 antibody validation summary (4 antibodies).
  5. MTA2-mediated inhibition of PTEN leads to pancreatic ductal adenocarcinoma carcinogenicity. Cell death & disease 2019 — PMC6393561.
  6. Prognostic, Immunological, and Mutational Analysis of MTA2 in Pan-Cancer and Drug Screening for Hepatocellular Carcinoma. Biomolecules 2023 — PMC10296612.
  7. Human Blastocyst Secreted microRNA Regulate Endometrial Epithelial Cell Adhesion. EBioMedicine 2015 — PMC4634783.
  8. MTA2 knockdown suppresses human osteosarcoma metastasis by inhibiting uPA expression. Aging 2024 — PMC11424574.
  9. PubMed PMID:9929979 — UniProt-cited evidence.
  10. PubMed PMID:11099047 — UniProt-cited evidence.
  11. PubMed PMID:14702039 — UniProt-cited evidence.