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MTA2 / Metastasis-associated protein MTA2 · IHC design guide
Design Immunohistochemistry for MTA2
Plan paraffin-section chromogenic IHC for MTA2 using its ubiquitous nuclear tissue pattern as a reference (HPA tissue IHC). Compare nuclear staining across matched cell populations, starting with the catalog antibody’s validated IHC-P conditions (datasheet A03073-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
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 | Ubiquitous nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | Nuclear staining across many 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 sections (standard IHC practice; not target-specific) | |
| Caveat | Ubiquitous staining leaves no obvious negative tissue (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended MTA2 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: A03073-1) with three published MTA2 immunohistochemistry 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. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 for the catalog antibody (datasheet: A03073-1); the published citrate protocols used other antibodies (PMC6393561; PMC10296612).
Section 2
What Is the Expected MTA2 Staining Pattern?
MTA2 should stain nuclei across many cell types in paraffin sections (UniProt O94776: nucleus; HPA tissue IHC: ubiquitous nuclear expression, Supported). High HPA staining is reported in adipocytes, glandular cells, hematopoietic cells, respiratory epithelial cells, glial cells and cerebellar granular-layer cells (HPA tissue IHC). MTA2 has no transmembrane segment or signal peptide (UniProt O94776 topology and processing).
| Nuclear staining in adipocytes, glandular cells or hematopoietic cells. | This matches the reported compartment and examples of high expression (HPA tissue IHC: ubiquitous nuclear expression; High in those cell types). Compare nuclei with adjacent cells and the counterstain when scoring (general IHC practice). |
| Predominantly cytoplasmic or membrane staining, with little nuclear signal. | This conflicts with MTA2's nuclear annotation and HPA's nuclear IHC profile (UniProt O94776: nucleus; HPA tissue IHC). Consider nonspecific staining or a detection artefact; compartment alone cannot identify its cause (general IHC practice). |
| Signal in a cell population expected to be negative solely because of its tissue identity. | Do not call this cross-reactivity on that basis: MTA2 is widely expressed, and HPA describes ubiquitous nuclear staining (UniProt O94776: tissue specificity; HPA tissue IHC). Cytoplasmic signal or staining tied to endogenous detection activity warrants a control-based check (general IHC practice). |
| Diffuse colour across nuclei, cytoplasm and tissue spaces. | A widespread haze obscures the nuclear pattern expected for MTA2 (HPA tissue IHC). Evaluate background with a no-primary control, then review blocking, washes and detection conditions (general IHC practice). |
| No nuclear signal in a known-positive tissue section. | This disagrees with HPA's High staining in the listed positive cell populations (HPA tissue IHC). Check that the relevant cells are present and assess the run with an established positive control before interpreting the section as negative (general IHC practice). |
💡Expected MTA2 appearanceCall a result positive when staining is predominantly nuclear in the relevant cells, with readily visible signal in HPA High examples; dominant cytoplasmic, membrane or diffuse background signal is suspect (HPA tissue IHC; UniProt O94776: nucleus; general IHC practice).
| Tissue and cell context | HPA reports ubiquitous nuclear expression and low tissue RNA specificity, with High IHC staining in eight listed tissue and cell contexts; its supplied list contains no negative tissue (HPA tissue IHC). Use a listed positive context to judge a run, without assuming unlisted cells are negative. |
| Antibody validation | IHC is Supported for HPA006214, CAB005315 and CAB017522; no IHC status is supplied for HPA072727 (HPA antibodies). These statuses support interpretation of the reported pattern but do not establish identical performance for every antibody or section. |
| IF/ICC Q: Where should signal appear? | In IF/ICC, expect nucleoplasmic signal; HPA calls that location Enhanced and lists ICC-IF images from A-431, U-251MG, U2OS, HAP1 and REH (HPA subcellular). Apply that location expectation to IF interpretation; this IHC section supplies no IF protocol. |
| Isoforms and epitope coverage | UniProt lists two MTA2 isoforms and domains at residues 1–315 (UniProt O94776: isoforms and domains). The supplied evidence gives no antibody epitope, so it cannot establish whether a particular antibody detects both isoforms or whether a domain affects staining. |
| Processing and retrieval | UniProt describes a 1–668 chain, with no signal peptide, propeptide or glycosylation sites (UniProt O94776: processing and glycosylation). Follow the catalog antibody's IHC-P retrieval instructions when available (general IHC practice); no MTA2-specific retrieval or fixation effect is supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control cells lack nuclear staining. | The expected signal is absent in a context reported as High (HPA tissue IHC); the failed step cannot be assigned from appearance alone (general IHC practice). | Confirm the relevant cells are on the section, then review the catalog antibody's IHC-P retrieval, dilution and detection instructions and the run controls (general IHC practice). |
| Cytoplasm or cell borders stain more strongly than nuclei. | That distribution conflicts with nuclear MTA2 (UniProt O94776: nucleus; HPA tissue IHC). | Compare with a no-primary control and nuclear counterstain; review blocking, washing and antibody conditions before scoring nuclear positivity (general IHC practice). |
| Broad brown background obscures cell boundaries. | Background can arise from detection chemistry or nonspecific binding; the image alone cannot distinguish them (general IHC practice). | Check a no-primary control and the detection system's endogenous-activity control, then review blocking and washes (general IHC practice). |
| Only one cell population stains within a mixed tissue. | MTA2 is widely expressed, but the supplied HPA data do not specify equal intensity in every cell (UniProt O94776: tissue specificity; HPA tissue IHC). | Identify cell types with the counterstain and compare nuclear signal in a listed HPA High cell population; avoid assigning negativity from tissue name alone (HPA tissue IHC; general IHC practice). |
| Signal appears in an unexpected cell population. | HPA's ubiquitous nuclear profile makes cell identity alone insufficient evidence of cross-reactivity (HPA tissue IHC). | First verify that signal is nuclear; use a no-primary control to assess detection background, then interpret the population in its tissue context (general IHC practice). |
| IF/ICC shows diffuse cytoplasmic fluorescence. | That pattern differs from HPA's Enhanced nucleoplasmic location (HPA subcellular). | Compare fluorescence with a nuclear counterstain and appropriate background controls; consult the separate IF/ICC guide for assay conditions (HPA subcellular; general IF practice). |
Experiment evidence
IHC & IF images showing MTA2 staining
A03073-1 · IHCIHC analysis of MTA2 using anti-MTA2 antibody (A03073-1). MTA2 was detected in paraffin-embedded section of human glioma tissue. Heat mediat
A03073-1 · IHCIHC analysis of MTA2 using anti-MTA2 antibody (A03073-1). MTA2 was detected in paraffin-embedded section of human glioma tissue. Heat mediat
A03073-1 · IFIF analysis of MTA2 using anti-MTA2 antibody (A03073-1). MTA2 was detected in immunocytochemical section of A549 cells. Enzyme antigen retri
M03073 · IHCIHC analysis of MTA2 using anti-MTA2 antibody (M03073). MTA2 was detected in a paraffin-embedded section of human cervical cancer tissue. He
M03073 · IHCIHC analysis of MTA2 using anti-MTA2 antibody (M03073). MTA2 was detected in a paraffin-embedded section of human cervical cancer tissue. He
A03073 · IHCImmunohistochemistry of PID in human small intestine tissue with PID antibody at 2.5 μg/ml.
A03073 · IFImmunocytochemistry staining of HeLa using PID antibody at 10 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000149480-MTA2/tissue/cerebral+cortex#imid_2079776
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000149480-MTA2/tissue/colon#imid_2079694
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000149480-MTA2/tissue/liver#imid_2079711
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000149480-MTA2/tissue/kidney#imid_2079704
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB017522; green = MTA2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB017522; green = MTA2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MTA2 IHC & IF
🧪Run bone marrow first and score nuclear staining in hematopoietic cells (HPA: High in bone marrow hematopoietic cells; UniProt O94776: nucleus). HPA detects MTA2 in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and treat any neighboring cell with a counterstained nucleus but no nuclear signal as an observational negative, not a validated MTA2-negative cell (HPA: no negative tissue rows; UniProt O94776: widely expressed).
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 and a concentration-matched rabbit IgG isotype slide; a matched MTA2 knockout specimen would provide a biological negative (selected-SKU caption: rabbit anti-MTA2 antibody; standard IHC control practice). Check endogenous peroxidase in bone marrow before interpreting DAB staining (HPA: bone marrow hematopoietic cells High; standard IHC practice).
⚠️Feasibility: The selected-SKU paraffin-section caption reports heat retrieval in EDTA at pH 8.0 and nuclear-compatible chromogenic detection, but it does not report the fixative; a target-specific fixation window or retrieval dependency is unreported (selected-SKU tissue-IHC caption; UniProt O94776: nucleus). The supplied evidence does not establish that frozen sections or IF are easier; HPA does show nucleoplasmic ICC-IF localization (HPA: nucleoplasm enhanced). In bone marrow, endogenous peroxidase can complicate DAB scoring (HPA: bone marrow hematopoietic cells High; 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
| 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 → |
Undetected expression · recommended negative controls
| 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. | ||||
Section 3
Advanced MTA2 IHC Tips
Troubleshoot MTA2 chromogenic IHC by checking retrieval, nuclear localisation, detection controls, and scoring before interpreting differences between samples.
What retrieval should I use when MTA2 staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A03073-1). The selected paraffin-section image used that condition, followed by 10% goat serum blocking and 1 μg/ml primary antibody overnight at 4°C (caption A03073-1). If nuclear staining remains weak, check heating consistency and section adhesion, then compare a small retrieval-condition series while keeping detection and exposure to chromogen constant (standard IHC practice). Evaluate any change against nuclear signal and tissue preservation, since MTA2 is nuclear and the reported tissue pattern is broadly nuclear (UniProt O94776; HPA: ubiquitous nuclear expression).
Could fixation explain inconsistent MTA2 staining between tissue blocks?
Target-specific sensitivity of MTA2 staining to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (caption A03073-1). Record each block's fixative and processing history, and compare blocks using the same pH 8.0 EDTA retrieval and detection settings (datasheet A03073-1; standard IHC practice). Include a section with previously acceptable nuclear staining in each run to distinguish a block problem from run variation (HPA: ubiquitous nuclear expression; standard IHC practice). Interpret differences cautiously when fixation histories differ, because variable processing can alter antigen accessibility and tissue morphology in paraffin-section IHC (standard IHC practice).
How should I assess apparent cytoplasmic MTA2 staining?
Score nuclear staining first: MTA2 is assigned to the nucleus, and the subcellular profile specifies nucleoplasm (UniProt O94776: subcellular location; HPA: enhanced nucleoplasmic localisation). Compare the suspect area with intact nuclei, the counterstain, and nearby well-preserved cells under the same chromogen development conditions (standard IHC practice). Diffuse cytoplasmic colour without corresponding nuclear signal warrants checks for nonspecific primary binding, excessive detection signal, and pigment or precipitate (standard IHC practice). Retest with a primary-antibody omission control and a dilution series around the caption's 1 μg/ml condition before assigning an unexpected compartment to MTA2 (caption A03073-1; standard IHC practice).
Can an MTA2 antibody distinguish isoforms or modified epitopes in IHC?
The record lists 2 MTA2 isoforms, but the supplied antibody caption does not identify its epitope or establish isoform selectivity (UniProt O94776: isoforms; caption A03073-1). MTA2 contains BAH, ELM2, and SANT domains and reported phosphorylation and acetylation sites, so epitope mapping would matter when interpreting differential staining (UniProt O94776: domains and modified residues). Do not assign a staining difference to a particular isoform or modification from chromogenic intensity alone (standard IHC interpretation). First confirm nuclear localisation, then seek antibody epitope documentation and an independently validated assay for the proposed molecular distinction (UniProt O94776: nucleus; standard IHC practice).
How can IF help investigate an ambiguous MTA2 IHC pattern?
Use IF as a separate localisation check: the reported MTA2 signal is nucleoplasmic, while this page's selected antibody evidence concerns paraffin-section chromogenic IHC (HPA: enhanced nucleoplasmic localisation; caption A03073-1). Multiplex MTA2 with an independently validated marker for the cell population under study and a nuclear counterstain, then compare signals cell by cell (standard IF practice). Choose a fluorophore channel with low measured tissue autofluorescence and include single-stain and unstained controls (standard IF practice). Because MTA2 is nuclear and lacks a transmembrane segment, use permeabilisation appropriate for access to nuclear epitopes and verify that it preserves morphology (UniProt O94776: location and topology; standard IF practice).
What should I check when DAB obscures MTA2 nuclei?
Separate diffuse colour from nuclear signal by examining a primary-antibody omission control and comparing development across matched sections (standard IHC practice; HPA: ubiquitous nuclear expression). The selected image used 10% goat serum block, 1 μg/ml primary antibody overnight at 4°C, and a biotin-based DAB detection sequence (caption A03073-1). For high background, check blocking, washes, primary concentration, chromogen development, and peroxidase blocking as parts of the general IHC workflow (standard IHC practice). With a biotin-based system, assess endogenous biotin as another possible source of signal before changing the interpretation of nuclear staining (caption A03073-1; standard IHC practice).
How should I quantify MTA2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the eligible cell population and score nuclear staining, consistent with the reported nuclear MTA2 distribution (UniProt O94776: nucleus; HPA: ubiquitous nuclear expression). An H-score can combine the percentage of nuclei in each intensity category; alternatively report percentage of positive nuclei or positive-nucleus density per mm² of viable tissue (standard IHC quantification practice). Normalize counts to all eligible nuclei in the same annotated compartment, and exclude folds, edges, and necrotic areas using predefined rules (standard IHC quantification practice). Keep retrieval, DAB development, counterstain, imaging, and positivity thresholds consistent across the comparison set (standard IHC practice).
How do I distinguish true MTA2 positivity from staining artefact?
Prioritize signal within intact nuclei: UniProt places MTA2 in the nucleus, and HPA reports a ubiquitous nuclear tissue pattern with supported reliability (UniProt O94776: location; HPA: tissue IHC). Broad staining across cell populations can therefore be plausible, but cell identity and nuclear boundaries still need confirmation on the counterstain (HPA: tissue IHC; standard IHC practice). Treat edge-concentrated colour, necrotic deposits, staining outside cells, and colour persisting after primary-antibody omission as possible artefacts (standard IHC practice). Check peroxidase blocking and the biotin-based detection controls when suspect signal could arise from endogenous enzyme activity or detection chemistry (caption A03073-1; standard IHC practice).
Boster reagents
Best MTA2 / Metastasis-associated protein MTA2 IHC Antibodies
Anti-MTA2 antibodies have IHC images from human glioma, cervical cancer and small intestine (catalog IHC captions), plus IF/ICC images from A549 and HeLa cells (catalog IF captions).
Real IHC data
Anti-MTA2/PID ® Antibody
Cat # A03073-1
Real IHC data
Anti-MTA2 Monoclonal Antibody
Cat # M03073
Real IHC data
Anti-PID MTA2 Antibody
Cat # A03073
A03073-1 has a paraffin-section IHC image from human glioma and an IF/ICC image from A549 cells (A03073-1 image captions); M03073 has a paraffin-section IHC image from human cervical cancer (M03073 image caption). A03073 has a human small-intestine IHC image and HeLa IF/ICC images (A03073 image captions).
Which to pick: For tissue IHC, choose A03073-1 for its human glioma paraffin-section example or M03073 for its human cervical cancer paraffin-section example (respective IHC captions); the fixative is unreported in both captions. For IF/ICC, choose A03073-1 for its A549 example or A03073 for its HeLa examples (respective IF captions). For planned mouse or rat work, A03073-1 and A03073 list both species as reactive, whereas the monoclonal M03073 lists human only (catalog reactivity and clone fields); the supplied images show human samples (image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O94776 (MTA2_HUMAN, Metastasis-associated protein MTA2).
- Human Protein Atlas. MTA2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. MTA2 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. MTA2 antibody validation summary (4 antibodies).
- MTA2-mediated inhibition of PTEN leads to pancreatic ductal adenocarcinoma carcinogenicity. Cell death & disease 2019 — PMC6393561.
- Prognostic, Immunological, and Mutational Analysis of MTA2 in Pan-Cancer and Drug Screening for Hepatocellular Carcinoma. Biomolecules 2023 — PMC10296612.
- Human Blastocyst Secreted microRNA Regulate Endometrial Epithelial Cell Adhesion. EBioMedicine 2015 — PMC4634783.
- MTA2 knockdown suppresses human osteosarcoma metastasis by inhibiting uPA expression. Aging 2024 — PMC11424574.
- PubMed PMID:9929979 — UniProt-cited evidence.
- PubMed PMID:11099047 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MTA2 (Metastasis-associated protein MTA2). Expected localisation Ubiquitous nuclear tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03073-1). Validated IHC antibody: A03073-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human glioma tissue; fixative not specified (datasheet A03073-1); 2. Fixation: Image fixative and duration unreported (datasheet A03073-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A03073-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03073-1); 8. Primary antibody: Rabbit anti-MTA2, 0.5-1μg/ml (datasheet A03073-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03073-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Ubiquitous staining leaves no obvious negative tissue (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). Evidence: UniProt O94776, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm., Datasheet A03073-1, PMC6393561, PMC10296612. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MTA2/immunohistochemistry-ihc-guide.