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DSTN / Destrin · IHC design guide
Design Immunohistochemistry for DSTN
Plan DSTN chromogenic IHC in paraffin sections around the reported general cytoplasmic staining pattern (HPA tissue IHC). High staining in colon glandular cells offers a positive reference, but antibody staining has low consistency with RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DSTN 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 | General cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M03849-2) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+2 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA expression show low consistency (HPA tissue IHC) | |
| Regulation | Specific expression regulators unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended DSTN IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 heat retrieval (datasheet: M03849-2). The published myocardial tissue protocol below provides a second IHC workflow (PMC12306564 methods).
| Sample | Paraffin-embedded mouse small intestine tissue; fixative not specified (datasheet M03849-2) |
| Fixation | Image fixative and duration unreported (datasheet M03849-2); 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 M03849-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M03849-2) |
| Primary antibody | Rabbit monoclonal (clone 19D11) anti-DSTN, 1:50 (datasheet M03849-2) |
| Primary incubation | Overnight at 4 °C (datasheet M03849-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M03849-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DSTN-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteTry EDTA pH 8.0 heat retrieval first (datasheet: M03849-2); the published protocol reports antigen retrieval without conditions (PMC12306564 methods).
Section 2
What Is the Expected DSTN Staining Pattern?
In paraffin-section IHC, expect mainly cytoplasmic DSTN staining in glandular cells of several tissues and in cerebellar Purkinje cells (HPA: general cytoplasmic expression; High in listed glandular cells and Purkinje cells). DSTN has no transmembrane segment (UniProt P60981: topology). HPA rates the tissue IHC profile Approved, while reporting low consistency between antibody staining and RNA expression; interpret the pattern with that qualification (HPA: reliability description).
| Clear cytoplasmic staining in colon or duodenal glandular cells, with visible cell boundaries and limited surrounding deposit. | This fits the reported IHC compartment and a cell population scored High in both tissues (HPA: general cytoplasmic expression; High in colon and duodenum glandular cells). Compare intensity among cells on the same section before treating a weak field as a biological difference (general IHC practice). |
| Strong nuclear-only staining, with little or no cytoplasmic signal in an otherwise readable section. | Nuclear-only signal does not match the reported general cytoplasmic IHC profile (HPA: tissue IHC profile). Check antibody dilution, detection chemistry, and a control section before assigning it to DSTN (general IHC practice). UniProt provides no DSTN subcellular annotation to support a nuclear call (UniProt P60981: subcellular location unannotated). |
| Prominent staining of adipocytes, hippocampal glial cells, or skeletal-muscle myocytes. | These populations were scored Not detected, so reproducible signal warrants scrutiny for cross-reactivity or endogenous detection activity (HPA: adipocytes, hippocampal glia, and skeletal-muscle myocytes Not detected; general IHC practice). A reported absence is a comparison point, not proof that every specimen must be blank (HPA: tissue IHC profile). |
| Brown deposit spreads across stroma, section edges, or the whole slide without resolving individual cell cytoplasm. | Treat a widespread, poorly localized deposit as background until controls show otherwise (general chromogenic IHC practice). It cannot by itself establish the cytoplasmic pattern reported for DSTN (HPA: general cytoplasmic expression); compare a no-primary control and inspect whether staining tracks tissue structures (general IHC practice). |
| No detectable signal in well-preserved colon or duodenal glandular cells. | A blank result conflicts with the reported High staining in those cells, but does not alone identify the cause (HPA: High in colon and duodenum glandular cells). Review positive-control performance, retrieval conditions, antibody dilution, and detection reagents as general IHC checks; no DSTN-specific retrieval requirement is supplied (general IHC practice). |
💡Expected DSTN appearanceCall a positive result when distinct cytoplasmic staining is evident in glandular cells of a reported High tissue or in Purkinje cells; diffuse deposit or dominant nuclear-only staining is suspect (HPA: general cytoplasmic expression; High in listed glandular cells and Purkinje cells; general IHC practice).
| Tissue and cell selection | Adrenal gland, appendix, colon, duodenum, endometrium, epididymis, and fallopian-tube glandular cells are scored High; Purkinje cells are also High (HPA: tissue IHC). Adipocytes, hippocampal glia, and skeletal-muscle myocytes are scored Not detected (HPA: tissue IHC). |
| Strength of the IHC evidence | The tissue profile is Approved, but HPA reports low consistency between antibody staining and RNA expression (HPA: reliability description). Its listed IHC antibody, CAB037094, is Approved; the supplied record does not label its IHC validation Enhanced (HPA: antibody validation). |
| Cellular compartment and topology | HPA describes general cytoplasmic tissue staining, and UniProt reports no transmembrane segment (HPA: tissue IHC profile; UniProt P60981: topology). Neither entry establishes a required membrane pattern in paraffin-section IHC (HPA: tissue IHC profile; UniProt P60981: subcellular location unannotated). |
| IF/ICC comparison? | HPA lists plasma membrane as the approved ICC-IF main location, with a caution that the result is based on antibodies targeting proteins from multiple genes (HPA: subcellular summary). Read that as a qualified IF observation; judge this IHC slide against HPA's cytoplasmic tissue profile (HPA: tissue IHC profile). |
| Isoforms and epitope interpretation | UniProt lists two DSTN isoforms and an ADF-H domain spanning residues 4–153 (UniProt P60981: isoforms and domains). Without an epitope map for the antibody, these facts do not predict which isoforms it recognizes or a retrieval setting (UniProt P60981: isoforms and domains; general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive glandular cells are blank. | The reported High signal is absent; the slide alone cannot distinguish assay failure from specimen variation (HPA: High in listed glandular cells; general IHC practice). | Check a positive-control section, detection reagents, antibody dilution, and retrieval settings; treat retrieval changes as general assay optimization (general IHC practice). |
| Most tissue structures stain similarly. | Uniform deposit obscures the expected cytoplasmic, cell-resolved profile (HPA: general cytoplasmic expression; general IHC practice). | Compare a no-primary control, inspect blocking and washes, and titrate antibody or detection reagents under the assay's standard controls (general IHC practice). |
| Red blood cells or other control-negative elements show chromogen. | Endogenous detection activity can contribute signal in chromogenic IHC (general IHC practice). | Use the appropriate endogenous-enzyme blocking step for the detection system and compare a no-primary control (general IHC practice). |
| Nuclei dominate while cytoplasm remains faint. | That distribution differs from the reported tissue IHC profile; UniProt has no subcellular annotation resolving it (HPA: general cytoplasmic expression; UniProt P60981: subcellular location unannotated). | Review counterstain, reagent concentration, and controls; avoid scoring nuclear-only deposit as expected DSTN staining (general IHC practice; HPA: tissue IHC profile). |
| Adipocytes or skeletal-muscle myocytes appear strongly positive. | Both are reported Not detected, making this an unexpected cell-type result that merits confirmation (HPA: adipocytes and skeletal-muscle myocytes Not detected). | Inspect morphology and a no-primary control, then compare staining in a reported High cell population on a control section (general IHC practice; HPA: tissue IHC). |
| IF images appear membrane-enriched while IHC looks cytoplasmic. | The modalities have different HPA observations, and the ICC-IF membrane assignment carries a multiple-gene antibody caution (HPA: subcellular summary; tissue IHC profile). | Score paraffin-section IHC using the reported cytoplasmic tissue pattern; consult the separate IF/ICC guide for that modality's interpretation (HPA: tissue IHC profile; subcellular summary). |
Experiment evidence
IHC & IF images showing DSTN staining
M03849-2 · IHCIHC analysis of Destrin using anti-Destrin antibody (M03849-2). Destrin was detected in a paraffin-embedded section of mouse small intestine
M03849-2 · IHCIHC analysis of Destrin using anti-Destrin antibody (M03849-2). Destrin was detected in a paraffin-embedded section of rat small intestine t
M03849-2 · IFImmunofluorescent analysis using the Antibody at 1:50 dilution.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000125868-DSTN/tissue/cerebral+cortex#imid_9098560
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000125868-DSTN/tissue/colon#imid_9076946
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000125868-DSTN/tissue/liver#imid_9076931
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000125868-DSTN/tissue/kidney#imid_9076940
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA077782; green = DSTN, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA077782; green = DSTN, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DSTN IHC & IF
🧪Run duodenum first and expect staining in its glandular cells (HPA: High in duodenal glandular cells); use adipose tissue as the negative comparator and assess its adipocytes (HPA: Not detected in adipocytes). On the duodenal slide, treat morphologically intact cells that show only counterstain as internal negative cells, but verify their status rather than assuming every nonglandular cell lacks DSTN.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show DSTN in A-431, OE19, U2OS, with annotated localisation: Plasma membrane (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide; a concentration-matched nonimmune rabbit IgG isotype control matched to the primary’s clonality when established; and DSTN knockout tissue or immunogen-peptide competition if available (caption: rabbit primary; clonality unreported). For the duodenal positive slide, quench endogenous peroxidase before HRP/DAB detection and block endogenous biotin if using avidin–biotin detection (caption: HRP/DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected M03849-2 tissue-IHC caption does not state a fixative (caption: fixative unreported). Its paraffin-section example uses heat retrieval in EDTA at pH 8.0; compare retrieved and unretrieved sections to assess retrieval dependence (caption: heat retrieval in EDTA, pH 8.0). The supplied evidence does not establish whether frozen sections or IF are easier; if using IF on duodenum, check tissue autofluorescence with a no-primary control (standard IF practice).
HPA tissue IHC evidence for DSTN
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular 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 → |
Section 3
Advanced DSTN IHC Tips
Troubleshoot DSTN staining in paraffin sections by checking retrieval, controls, cellular distribution and scoring before interpreting differences between specimens.
Which retrieval conditions should I try first for DSTN IHC?
Start with heat-mediated EDTA retrieval at pH 8.0 for paraffin sections (datasheet M03849-2). The selected mouse small-intestine image used this retrieval before incubation with 1:50 primary antibody overnight at 4°C (caption M03849-2). If staining is weak, compare heating and cooling intervals on adjacent sections while keeping antibody dilution, section thickness and DAB development consistent (standard IHC practice). If those adjustments fail, test a citrate-buffer fallback on matched sections and record its pH, signal and morphology (standard IHC practice). Include a no-primary control to check whether retrieval increases nonspecific staining (standard IHC practice).
Could fixation explain weak or uneven DSTN staining?
DSTN-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (caption M03849-2). Record each specimen’s fixative, fixation duration, processing schedule and section age before comparing staining across blocks (standard IHC practice). For a controlled comparison, stain adjacent sections together using heat-mediated EDTA at pH 8.0 and the same detection conditions (datasheet M03849-2; standard IHC practice). If one block stains poorly, check morphology and a control antigen known to survive its processing before changing DSTN antibody concentration (standard IHC practice). Do not treat staining differences alone as evidence of a DSTN-specific fixation effect (caption M03849-2).
How should I evaluate cytoplasmic versus membrane DSTN staining?
For chromogenic tissue IHC, assess cytoplasmic signal first because the tissue profile describes general cytoplasmic expression (HPA: tissue IHC profile). DSTN has no transmembrane segment, although it binds actin filaments and monomers (UniProt P60981 topology and function). A subcellular ICC/IF entry reports approved plasma-membrane localisation, with a caution that its antibodies target proteins from multiple genes (HPA: subcellular). Score crisp membrane accentuation separately from diffuse cytoplasmic staining, and compare both patterns with no-primary and tissue controls (standard IHC practice). Do not assign a membrane rim to DSTN solely from that ICC/IF entry; evaluate whether the same pattern recurs across intact cells (HPA: subcellular; standard IHC practice).
Can this stain distinguish DSTN isoforms or modified forms?
DSTN has 2 listed isoforms, but the supplied IHC caption does not identify an antibody epitope or establish isoform selectivity (UniProt P60981 isoforms; caption M03849-2). Its ADF-H domain spans residues 4–153, and reported modifications include phosphoserine at residue 3 (UniProt P60981 domains and modified residues). Without an epitope map and validation against each form, interpret chromogenic signal as antibody-reactive DSTN rather than an isoform or modification state (standard IHC interpretation). Keep retrieval and detection identical when comparing specimens, since changing either can alter apparent staining independently of biological abundance (standard IHC practice). Request epitope information or use a separately validated reagent before making form-specific claims (standard IHC practice).
How can I compare DSTN IHC with a multiplex IF experiment?
Treat IF/ICC as a separate assay: the supplied 1:50 dilution and EDTA pH 8.0 retrieval belong to the selected paraffin-section IHC image (caption M03849-2). Pair DSTN with a validated marker for the cell population being assessed, and check each channel independently before interpreting colocalisation (standard IF practice). Choose fluorophores and exposure settings after inspecting unstained tissue for autofluorescence, especially where broad background could obscure weak signal (standard IF practice). If the antibody recognises an intracellular epitope, optimise permeabilisation for access to that side of the membrane; the supplied evidence does not map this antibody’s epitope (standard IF practice; caption M03849-2). Compare compartment patterns cautiously because tissue IHC reports cytoplasmic expression while the ICC/IF entry reports plasma-membrane localisation with an antibody-specificity caution (HPA: tissue IHC and subcellular).
What should I check when DAB staining is widespread?
The selected image used goat-serum blocking, a peroxidase-linked secondary antibody and DAB development (caption M03849-2). First compare a no-primary section with the test section to identify detection-system signal that persists without DSTN primary antibody (standard IHC practice). Include a peroxidase-blocking step and monitor DAB development, since endogenous enzyme activity or excessive development can produce misleading brown deposits (standard IHC practice). If background persists, titrate the primary antibody below the caption’s 1:50 condition and compare matched sections with identical retrieval and exposure to chromogen (caption M03849-2; standard IHC practice). Judge improvement by preserved cellular staining and clean surrounding tissue, rather than by reduced overall darkness alone (standard IHC practice).
How should I quantify DSTN across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell compartment and analysis region before scoring, because the tissue IHC profile describes general cytoplasmic expression (HPA: tissue IHC profile). Record the percentage of positive cells and staining intensity, then calculate an H-score with intensity categories 0–3 if that scale is used consistently (standard IHC practice). For spatial comparisons, positive-cell density per mm² can complement the H-score, with necrotic and damaged areas excluded by a prespecified rule (standard IHC practice). Normalise to the number of eligible cells or viable tissue area within the same annotated cell population, rather than comparing raw stained area across different specimens (standard IHC practice). Keep retrieval, imaging, thresholds and scoring blinded and consistent across batches (standard IHC practice).
How do I distinguish genuine DSTN signal from artefact?
Expect a reproducible cellular pattern compatible with the reported general cytoplasmic tissue staining, while assessing any membrane accentuation separately (HPA: tissue IHC profile; HPA: subcellular). Compare intact cells with no-primary controls and examine whether brown signal instead follows tissue edges, folds or necrotic debris (standard IHC practice). Check endogenous peroxidase blocking when staining persists without primary antibody, since the selected IHC image uses peroxidase detection and DAB (caption M03849-2; standard IHC practice). Do not call every unstained cell a technical failure: adipocytes, hippocampal glial cells and skeletal-muscle myocytes were reported as not detected in the supplied tissue profile (HPA: tissue IHC). Treat isolated unexpected patterns cautiously because that profile reports low consistency between antibody staining and RNA expression (HPA: reliability description).
Boster reagents
Best DSTN / Destrin IHC Antibodies
The catalog antibody has IHC images from paraffin sections of mouse and rat small intestine (IHC captions); its IF captions give dilutions but no sample identity (IF captions).
Real IHC data
Anti-Destrin Rabbit Monoclonal Antibody
Cat # M03849-2
M03849-2 has IHC images of paraffin sections from mouse and rat small intestine (IHC captions). Its IF images list 1:50 and 1:150 dilutions without identifying specimens (IF captions); human, mouse, and rat reactivity is listed in the catalog (catalog: reactivity).
Which to pick: For tissue IHC, choose M03849-2: its caption documents paraffin sections, EDTA retrieval at pH 8.0, and a 1:50 primary dilution; the fixative is unreported (M03849-2 IHC caption). For IF/ICC, the same rabbit monoclonal is listed for both applications, although its IF captions do not identify the samples (catalog: host, clonality, applications; IF captions). For work across species, the catalog lists human, mouse, and rat reactivity, with pictured IHC evidence for mouse and rat only (catalog: reactivity; IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P60981 (DEST_HUMAN, Destrin).
- Human Protein Atlas. DSTN tissue IHC expression (reliability: Approved).
- Human Protein Atlas. DSTN subcellular location (ICC-IF): Localized to the plasma membrane. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. DSTN antibody validation summary (2 antibodies).
- Single-cell disulfidptosis regulator patterns guide intercellular communication of tumor microenvironment that contribute to kidney renal clear cell carcinoma progression and immunotherapy. Frontiers in immunology 2024 — PMC10824999.
- Disulfidptosis-Related Genes as Novel Biomarkers and Therapeutic Targets in Dilated Cardiomyopathy. Journal of inflammation research 2025 — PMC12306564.
- Differences in corneal phenotypes between destrin mutants are due to allelic difference and modified by genetic background. Molecular vision 2012 — PMC3298451.
- Abnormal epithelial homeostasis in the cornea of mice with a destrin deletion. Molecular vision 2008 — PMC2573731.
- PubMed PMID:8399167 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:11780052 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DSTN (Destrin). Expected localisation General cytoplasmic staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M03849-2). Validated IHC antibody: M03849-2 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse small intestine tissue; fixative not specified (datasheet M03849-2); 2. Fixation: Image fixative and duration unreported (datasheet M03849-2); 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 M03849-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M03849-2); 8. Primary antibody: Rabbit monoclonal (clone 19D11) anti-DSTN, 1:50 (datasheet M03849-2); 9. Primary incubation: Overnight at 4 °C (datasheet M03849-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA expression show low consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unknown (UniProt); Regulation: Specific expression regulators unreported (UniProt). Evidence: UniProt P60981, HPA tissue IHC · Approved, HPA ICC-IF · plasma membrane. Caution: Based on antibodies targeting proteins…, Datasheet M03849-2, PMC12306564. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DSTN/immunohistochemistry-ihc-guide.