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DMTN / Dematin · IHC design guide
Design Immunohistochemistry for DMTN
Plan paraffin-section DMTN IHC around cytoplasmic staining in several tissues and red blood cell positivity (HPA tissue IHC). Start the catalog antibody at 1:100–1:300 (datasheet: A07422), and interpret results cautiously because antibody staining and RNA show low consistency (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DMTN 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 | Cytoplasmic staining in tissue (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic signal in several tissues; red blood cells positive (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Appendix+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining and RNA show low consistency (HPA tissue IHC) | |
| Regulation | Brain-enhanced RNA expression (HPA tissue RNA) | |
| Isoform / epitope | Four isoforms; isoform-specific staining is unresolved (UniProt) |
Section 1
Recommended DMTN IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with one published DMTN IHC protocol (PMC6277997).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A07422) |
| Fixation | Image fixative and duration unreported (datasheet A07422); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| 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-DMTN, 1:100-1:300 (datasheet A07422) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DMTN-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in several tissue. Additional positivity in red blood cells. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen-retrieval rule); the article does not specify retrieval conditions (PMC6277997).
Section 2
What Is the Expected DMTN Staining Pattern?
DMTN is a cytoplasmic and cytoskeletal protein that also associates with membranes, including the erythrocyte spectrin–actin junction; it has no transmembrane segment (UniProt Q08495 topology/localization). In tissue IHC, expect cytoplasmic staining in selected cells and possible red blood cell positivity (HPA: tissue IHC profile). Treat the pattern as provisional: HPA rates tissue staining Uncertain because antibody staining and RNA expression show low consistency (HPA: reliability).
| Cytoplasmic staining in bronchial respiratory epithelium, with red blood cell positivity. | This fits the reported tissue profile: bronchial respiratory epithelial cells show Medium staining, and HPA notes additional red blood cell positivity (HPA: bronchus; tissue IHC profile). Assess epithelial cytoplasm separately from blood-filled spaces; erythrocyte-associated DMTN is biologically plausible (UniProt Q08495 localization). |
| Signal follows cell borders or processes as well as cytoplasm, without a dominant nuclear pattern. | A membrane-associated component can fit DMTN’s cytoskeletal and erythrocyte membrane localization, although HPA describes the tissue pattern broadly as cytoplasmic (UniProt Q08495 localization; HPA: tissue IHC profile). A predominantly nuclear result lacks support from either supplied localization record and warrants comparison with controls (UniProt Q08495 localization; HPA: tissue IHC profile). |
| Strong staining appears in appendix glandular cells while the expected comparison tissue stains weakly or not at all. | HPA reports appendix glandular cells as Not detected; this contrast raises concern for cross-reactivity or detection-system activity (HPA: appendix; general IHC practice). It is a warning, not proof of antibody failure, because the HPA tissue IHC profile itself is rated Uncertain (HPA: reliability). |
| Weak, diffuse chromogen covers stroma, lumina and many unrelated cells without clear boundaries. | A widespread haze does not resemble the reported cytoplasmic, cell-associated pattern (HPA: tissue IHC profile). Compare the no-primary control and examine blocking, washes and detection conditions before assigning that signal to DMTN (general IHC practice). |
| No convincing signal appears in bronchial respiratory epithelial cells or kidney glomerular cells. | Both are reported at Medium staining and can serve as comparison areas (HPA: bronchus; kidney). Check tissue integrity, the positive control and the detection run before interpreting absence as biological; the HPA result is Uncertain and does not guarantee positivity in every specimen (HPA: reliability; general IHC practice). |
💡Expected DMTN appearanceCall a result plausible when selected cells show discernible cytoplasmic staining, such as Medium signal in bronchial respiratory epithelium, with possible red blood cell positivity; isolated nuclear staining or uniform diffuse haze is suspect (HPA: bronchus; tissue IHC profile; UniProt Q08495 localization).
| Cell type and compartment | Compare like cells: HPA reports Medium staining in bronchial respiratory epithelial cells and kidney glomerular cells, but Not detected in appendix glandular cells (HPA: bronchus; kidney; appendix). UniProt places DMTN in cytosol, cytoskeleton and membrane-associated sites, with no transmembrane segment (UniProt Q08495 topology/localization). |
| Confidence in the tissue pattern | HPA rates tissue IHC Uncertain owing to low consistency between antibody staining and RNA expression; both listed rabbit polyclonal antibodies have Uncertain IHC validation (HPA: reliability; antibodies HPA024290/HPA058487). Use morphology and controls when interpreting a deviation (general IHC practice). |
| Isoforms and epitope interpretation | UniProt lists 4 DMTN isoforms and an HP domain at residues 337–405 (UniProt Q08495 isoforms/domains). The supplied sources do not map the antibody epitope, so staining cannot establish which isoform was detected or whether the domain is represented. |
| Modification and processing | UniProt lists 19 modified residues, including phosphoserines, and a chain spanning residues 1–405, with no signal peptide or propeptide (UniProt Q08495 modifications/processing). These facts alone do not predict staining strength, antigen retrieval needs or epitope accessibility. |
| Chromogenic detection in blood-rich areas | Red blood cells may contain target-associated signal (HPA: tissue IHC profile; UniProt Q08495 localization). In peroxidase-based IHC, endogenous activity can also produce chromogen, so judge blood-rich areas against an appropriate no-primary control (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Bronchial respiratory epithelium is negative. | The run may have a staining or detection problem, or this specimen may differ from the reported Medium pattern (HPA: bronchus; reliability; general IHC practice). | Review morphology and a concurrent positive control, then check the antibody and detection steps before calling the sample negative (general IHC practice). |
| Appendix glandular cells stain more strongly than the expected positive comparison cells. | That reverses HPA’s Not detected appendix glandular pattern and may reflect cross-reactivity or background (HPA: appendix; general IHC practice). | Compare the same cell types across sections, inspect a no-primary control and avoid treating this contrast alone as proof of DMTN expression (general IHC practice; HPA: reliability). |
| Red blood cell-rich spaces dominate the slide. | HPA reports red blood cell positivity, and erythrocyte membrane association is described by UniProt; peroxidase-based detection can add endogenous chromogen (HPA: tissue IHC profile; UniProt Q08495 localization; general IHC practice). | Score tissue cells separately from blood and compare with a no-primary control; assess endogenous peroxidase blocking if the control also develops chromogen (general IHC practice). |
| Most nuclei stain while cytoplasm is faint. | Predominantly nuclear staining is unsupported by the supplied DMTN localization and tissue IHC profile (UniProt Q08495 localization; HPA: tissue IHC profile). | Check the no-primary control, counterstain and detection conditions; require a convincing cell-associated cytoplasmic pattern before attributing nuclear color to DMTN (general IHC practice). |
| Diffuse background obscures cell boundaries. | Non-specific binding or insufficient washing can create a broad chromogenic haze (general IHC practice). | Compare the no-primary control and review blocking, washes and antibody concentration; reassess only interpretable, cell-associated signal (general IHC practice). |
| Can IF/ICC localization be inferred from these images? | HPA provides no main ICC-IF location and lists 0 cell lines with ICC-IF images (HPA: subcellular record). | Use UniProt’s cytoplasmic, cytoskeletal and membrane-associated localization only as a hypothesis; confirm any IF/ICC pattern with controls in the separate IF/ICC design (UniProt Q08495 localization; HPA: subcellular record; general IF practice). |
Experiment evidence
IHC & IF images showing DMTN staining
A07422 · IHCImmunohistochemistry analysis of paraffin-embedded human brain tissue, using Dematin Antibody. The picture on the right is blocked with the
A07422 · IFImmunofluorescence analysis of HUVEC cells, using Dematin Antibody. The picture on the right is blocked with the synthesized peptide.
A30444 · IHCImmunohistochemistry analysis of paraffin-embedded human brain tissue, using Dematin Antibody. The picture on the right is blocked with the
A30444 · IFImmunofluorescence analysis of HUVEC cells, using Dematin Antibody. The picture on the right is blocked with the synthesized peptide.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000158856-DMTN/tissue/cerebral+cortex#imid_6435288
HPA · LungHuman Protein Atlas IHC, Lung. Source: https://www.proteinatlas.org/ENSG00000158856-DMTN/tissue/lung#imid_6437839
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000158856-DMTN/tissue/colon#imid_6437894
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000158856-DMTN/tissue/liver#imid_6437895
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DMTN IHC & IF
🧪Run cerebral cortex first: neuropil should stain at a Medium level (HPA: cerebral cortex, neuropil); use appendix glandular cells as the negative tissue (HPA: appendix glandular cells, Not detected). On the positive slide, nuclei should lack specific DMTN staining (UniProt Q08495: cytoplasmic and membrane localization).
Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)
Negative control tissue: Appendix (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for DMTN; derive a cell-line control from the positive tissue's cell type (Adipocytes) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species, immunoglobulin class and concentration (standard IHC practice). Use peptide blocking as a biological specificity control (A07422 tissue-IHC caption: signal blocked with synthesized peptide); quench endogenous peroxidase and inspect vascular erythrocytes separately because DMTN localizes to their membrane (standard chromogenic IHC practice; UniProt Q08495: erythrocyte membrane localization).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A07422 paraffin-section caption does not state a fixative (A07422 tissue-IHC caption). Retrieval dependence is unreported; optimize antigen retrieval empirically for paraffin sections (standard IHC practice). Neither frozen sections nor IF can be identified as easier from the supplied evidence; in cerebral cortex, distinguish neuropil staining from DMTN-positive vascular erythrocytes (HPA: cerebral cortex, neuropil Medium; UniProt Q08495: erythrocyte membrane localization).
HPA tissue IHC evidence for DMTN
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low 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 | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuropil | Medium | Protein (IHC) | HPA → |
| Colon | Endothelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced DMTN IHC Tips
Troubleshoot DMTN staining in paraffin sections by checking retrieval, compartment, cell identity, controls, and scoring before interpreting intensity.
How should I retrieve DMTN when paraffin-section staining is weak?
Use citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval specification). Cool sections gradually, keep them hydrated, and compare staining with a matched control section so retrieval changes can be judged consistently (standard IHC practice). DMTN occurs in cytoskeletal, cytosolic, and membrane-associated compartments, so assess both cellular staining and tissue preservation (UniProt Q08495 localisation). If signal remains weak, optimise retrieval duration on adjacent sections while holding antibody and detection conditions constant (standard IHC practice). The selected antibody has a paraffin-embedded brain image with peptide blocking, but its caption supplies no retrieval conditions (A07422 caption).
Could fixation explain weak or uneven DMTN staining?
Target-specific fixation sensitivity is unknown because the supplied DMTN evidence gives no fixation comparison or fixation condition for this antibody (A07422 caption; supplied evidence). Record the fixative, fixation duration, processing history, and section thickness for each specimen before comparing staining (standard IHC practice). Uneven preservation can change morphology and antigen accessibility, so examine adjacent sections and include similarly processed controls (standard IHC practice). If staining differs across specimens, optimise retrieval and antibody concentration using sections with matched processing before attributing the difference to DMTN expression (standard IHC practice). The paraffin-embedded brain caption does not identify a fixative (A07422 caption).
Which compartments should show credible DMTN staining?
Expect cytoplasmic or cytoskeletal staining, with possible membrane-associated or perinuclear signal, because these locations are recorded for DMTN (UniProt Q08495 localisation). In erythrocytes, DMTN localises to the spectrin–actin junction at the plasma membrane (UniProt Q08495 localisation). Evaluate cell boundaries and morphology with the counterstain before calling a narrow rim positive (standard IHC practice). HPA describes cytoplasmic staining across several tissues and additional red blood cell positivity, but assigns its tissue IHC profile uncertain reliability (HPA tissue IHC). Predominantly nuclear staining deserves investigation with peptide blocking or an independent antibody before it is reported as DMTN (UniProt Q08495 localisation; standard IHC practice).
Can this antibody distinguish DMTN isoforms or phosphorylation states in sections?
Do not assign an IHC pattern to one isoform without an isoform-resolved epitope map or direct validation: DMTN has 4 listed isoforms (UniProt Q08495 isoforms; standard IHC practice). Its C-terminal HP domain spans residues 337–405, while listed phosphorylation sites include serines 16, 18, 26, 92, and 96 (UniProt Q08495 domains; modified residues). Those features may matter if an antibody recognises a mapped region, but this payload does not map the catalog antibody’s epitope (supplied evidence). The selected brain image includes a synthesized-peptide blocking comparison, which supports investigation of binding specificity without identifying an isoform or phosphorylation state (A07422 caption; standard IHC practice). Compare an independent epitope or perturbation control before making either claim (standard IHC practice).
How can I assess DMTN localisation by multiplex IF?
Treat IF as a separate optimisation because the supplied catalog image documents paraffin-section chromogenic IHC, not IF performance (A07422 caption). Pair DMTN with a validated marker for the expected cell population, and inspect single-channel images before interpreting overlap (standard IF practice). Choose fluorophores after measuring tissue autofluorescence, placing the weaker signal in a channel with better separation from background (standard IF practice). DMTN has no transmembrane segment and is reported in cytosolic, cytoskeletal, and membrane-associated locations, so choose permeabilisation according to the mapped epitope’s accessibility (UniProt Q08495 topology; localisation; standard IF practice). Its antibody epitope is unspecified here; test permeabilised and unpermeabilised controls before interpreting a membrane rim (supplied evidence; standard IF practice).
What should I check when DMTN IHC looks diffusely brown?
First compare no-primary and peptide-blocked controls with the test section; the selected brain image includes a peptide-blocked comparison (A07422 caption; standard IHC practice). Block endogenous peroxidase before chromogenic detection and check whether blood-rich regions retain colour in the no-primary control (standard IHC practice). Red blood cell positivity can also be biologically plausible for DMTN, so distinguish cellular staining from detection background by morphology and controls (HPA tissue IHC; UniProt Q08495 localisation). If haze persists, titrate the primary antibody and adjust blocking or washing while keeping retrieval constant (standard IHC practice). Score only interpretable cells in intact tissue, excluding folds and damaged edges (standard IHC practice).
How should I quantify heterogeneous DMTN staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because DMTN can appear cytoplasmic or membrane-associated and red blood cells may stain (UniProt Q08495 localisation; HPA tissue IHC). For cellular regions, report the percentage of positive eligible cells and, when intensity is reproducible, an H-score from intensity-weighted percentages (standard IHC practice). For sparse positive cells, report counts per mm² of viable, evaluable tissue; normalise each count to the measured area or eligible cell count (standard IHC practice). Keep retrieval, detection, counterstain, image settings, and positivity thresholds consistent across comparison groups (standard IHC practice). Record red blood cells separately from the target population to avoid inflating tissue-cell scores (HPA tissue IHC; standard IHC practice).
When is a DMTN-positive pattern convincing rather than artefactual?
A convincing result has interpretable cellular morphology, a plausible cytoplasmic, cytoskeletal, or membrane-associated pattern, and appropriate control behaviour (UniProt Q08495 localisation; standard IHC practice). In brain sections, identify the stained structure before equating signal with a cell type; HPA reports medium staining in cerebral cortex neuropil (HPA tissue IHC). Compare peptide-blocked and no-primary sections, and inspect section edges, folds, and necrotic areas for misleading colour (A07422 caption; standard IHC practice). Persistent colour after omitting primary antibody can indicate endogenous enzyme activity or detection background (standard IHC practice). Interpret tissue comparisons cautiously because HPA rates its DMTN tissue IHC profile uncertain owing to low staining–RNA consistency (HPA tissue IHC).
Boster reagents
Best DMTN / Dematin IHC Antibodies
Anti-DMTN antibodies have paraffin-section IHC images from human brain and IF images from HUVEC cells (catalog image captions); both list human and mouse reactivity (catalog applications/reactivity).
Real IHC data
Anti-Dematin DMTN Antibody
Cat # A07422
Real IHC data
Anti-Dematin DMTN Antibody
Cat # A30444
A07422 lists IHC and IF, with human brain paraffin-section IHC and HUVEC IF images (A07422 applications and image captions). A30444 lists IHC, IF and ICC, with human brain paraffin-section IHC and HUVEC IF images (A30444 applications and image captions).
Which to pick: For tissue IHC, either SKU has its own human brain paraffin-section image (A07422 and A30444 IHC image captions); the fixative is unreported in both captions. For IF, either lists IF and has its own HUVEC image; choose A30444 if ICC is required because ICC appears in its application list (A07422 and A30444 applications and IF image captions). For human or mouse samples, both are listed as rabbit polyclonal antibodies reactive with those species (A07422 and A30444 catalog payloads).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q08495 (DEMA_HUMAN, Dematin).
- Human Protein Atlas. DMTN tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. DMTN subcellular location (ICC-IF): Highest expression in HEL: 191.1 nTPM.
- Human Protein Atlas. DMTN antibody validation summary (2 antibodies).
- Hypermethylation of DMTN promotes the metastasis of colorectal cancer cells by regulating the actin cytoskeleton through Rac1 signaling activation. Journal of experimental & clinical cancer research : CR 2018 — PMC6277997.
- PubMed PMID:8341682 — UniProt-cited evidence.
- PubMed PMID:7615546 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DMTN (Dematin). Expected localisation Cytoplasmic staining in tissue (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A07422 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Appendix. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet A07422); 2. Fixation: Image fixative and duration unreported (datasheet A07422); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-DMTN, 1:100-1:300 (datasheet A07422); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Caveat: Antibody staining and RNA show low consistency (HPA tissue IHC); Isoform / epitope: Four isoforms; isoform-specific staining is unresolved (UniProt); Regulation: Brain-enhanced RNA expression (HPA tissue RNA). Evidence: UniProt Q08495, HPA tissue IHC · Uncertain, HPA cell-line RNA · Highest expression in HEL: 191.1 nTPM, Datasheet A07422, PMC6277997. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DMTN/immunohistochemistry-ihc-guide.