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DCTN1 / Dynactin subunit 1 · IHC design guide
Design Immunohistochemistry for DCTN1
Plan chromogenic DCTN1 IHC on paraffin sections with the catalog antibody at 2–5 μg/mL (datasheet: A02175-2). Compare staining with HPA’s high staining in fallopian tube glandular cells and undetected staining in lymph node germinal center cells (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DCTN1 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 and nuclear staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Cells in most tissues show cytoplasmic and nuclear staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02175-2) | |
| Positive control | Fallopian tube+4 more · see all | |
| Negative control | Lymph node+1 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 | Germinal center cells may lack detectable staining (HPA tissue IHC) | |
| Regulation | No expression regulator specified (UniProt) | |
| Isoform / epitope | 6 isoforms; antibody epitope coverage is unspecified (UniProt) |
Section 1
Recommended DCTN1 IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet). Published examples include direct DCTN1 staining in human paraffin sections (PMC3898179) and FLAG-based detection of human DCTN1 in mouse free-floating sections (PMC12730482).
| Sample | Paraffin-embedded human ovarian adenocarcinoma tissue; fixative not specified (datasheet A02175-2) |
| Fixation | Image fixative and duration unreported (datasheet A02175-2); 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 A02175-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02175-2) |
| Primary antibody | Rabbit anti-DCTN1, 2-5μg/ml (datasheet A02175-2) |
| Primary incubation | Overnight at 4 °C (datasheet A02175-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02175-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DCTN1-positive staining in glandular cells of fallopian tube (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA, pH 8.0, for the catalog antibody (datasheet A02175-2). The citrate retrieval reported in PMC3898179 was for DCTN3, not DCTN1 (PMC3898179 methods).
Section 2
What Is the Expected DCTN1 Staining Pattern?
DCTN1 is a non-transmembrane protein associated with the cytoplasm, cytoskeleton, centrosome, spindle, nuclear envelope and cell cortex (UniProt Q14203 topology and subcellular location). In tissue IHC, expect cytoplasmic and sometimes nuclear staining across many cell types, including glandular and neuronal cells (HPA: tissue IHC profile). HPA rates its tissue staining “Approved,” with medium consistency against RNA data and external verification pending (HPA: tissue IHC reliability).
| Cytoplasmic staining in fallopian tube or gallbladder glandular cells, with possible nuclear staining. | This fits the reported tissue pattern: both glandular populations stain at a high level, and the overall profile includes cytoplasmic and nuclear expression (HPA: tissue IHC). Compare cells within the section when judging distribution (general IHC practice). |
| Staining is confined to cell surfaces or extracellular spaces, with no convincing intracellular signal. | Treat this as a compartment mismatch and investigate artefact: DCTN1 has no transmembrane segment, while its reported locations are intracellular (UniProt Q14203 topology and subcellular location). Nuclear staining alone is not a mismatch because HPA reports nuclear expression (HPA: tissue IHC profile). |
| A strong reaction dominates a cell population reported as undetected, while expected positive cells are unstained. | Consider cross-reactivity or endogenous detection activity and check the assay controls (general IHC practice). HPA reports no detection in lymph-node germinal-center cells and spleen red-pulp cells, but these observations are assay-specific, not universal negative controls (HPA: tissue IHC). |
| Diffuse color covers cells and surrounding tissue without clear cellular boundaries. | This is difficult to score as DCTN1 because the reported pattern is cellular (HPA: tissue IHC profile). Review background in a control lacking primary antibody, blocking, washes and chromogen development; these are general IHC checks, not documented DCTN1-specific effects (general IHC practice). |
| No signal appears in fallopian tube or gallbladder glandular cells. | First check section quality, controls, retrieval and detection before calling DCTN1 absent (general IHC practice). HPA reports high staining in these cells, but its tissue evidence has medium consistency with RNA data and awaits external verification (HPA: tissue IHC and reliability). |
💡Expected DCTN1 appearanceCall a result positive when intracellular staining, chiefly cytoplasmic with possible nuclear signal, is visible in glandular cells of fallopian tube or gallbladder at a high relative level (HPA: tissue IHC); an isolated surface or extracellular deposit is a compartment mismatch (UniProt Q14203 topology and subcellular location).
| Tissue and cell selection | HPA reports high staining in fallopian-tube and gallbladder glandular cells; medium staining in several other listed populations, including caudate neurons; and low staining in hippocampal neurons (HPA: tissue IHC). Select comparison cells by their reported level rather than treating all tissues as equally bright (general IHC practice). |
| Compartment used for scoring | HPA summarizes tissue staining as cytoplasmic and nuclear in most tissues (HPA: tissue IHC profile). UniProt also lists cytoskeletal, centrosomal, spindle, nuclear-envelope and cortical locations (UniProt Q14203 subcellular location). A fine structure seen in cells need not resolve as a distinct feature in chromogenic tissue IHC (general IHC practice). |
| Antibody evidence | Four listed antibodies have an “Approved” IHC status; the tissue profile has medium consistency with RNA expression and awaits external verification (HPA: antibodies and tissue IHC reliability). An Approved label supports use as a starting point but does not establish that every positive deposit is specific (general IHC practice). |
| Isoforms and epitope | UniProt lists six DCTN1 isoforms and a CAP-Gly domain at residues 48–90 (UniProt Q14203 isoforms and domains). The supplied evidence does not locate any catalog-antibody epitope, so isoform coverage and any retrieval dependence cannot be inferred from it (UniProt Q14203; HPA: supplied antibody records). |
| Processing and fixation evidence | UniProt lists one chain spanning residues 1–1278, with no signal peptide or propeptide and no reported glycosylation sites (UniProt Q14203 processing and glycosylation). Neither source supplied here reports a DCTN1-specific fixation effect; determine retrieval conditions with assay controls (UniProt Q14203; HPA: supplied records; general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected high-staining glandular cells are blank. | Section or detection failure is possible (general IHC practice); high staining is reported for these cells, though HPA verification remains pending (HPA: tissue IHC and reliability). | Check a positive section and detection controls, then review retrieval, antibody dilution and incubation against the chosen antibody's validated IHC conditions (general IHC practice). |
| Only a surface rim or extracellular material stains. | That distribution conflicts with the intracellular locations and absence of a transmembrane segment (UniProt Q14203 topology and subcellular location). | Inspect a control lacking primary antibody and reassess chromogen deposits and section morphology before scoring the rim as positive (general IHC practice). |
| Lymph-node germinal-center or spleen red-pulp cells stain strongly. | HPA reports these populations as not detected; cross-reactivity or endogenous detection activity is possible (HPA: tissue IHC; general IHC practice). | Compare a control lacking primary antibody and check endogenous-activity blocking; interpret the HPA negative observations within their assay limits (general IHC practice; HPA: tissue IHC). |
| Color is widespread, hazy or similar in cells and stroma. | High background can obscure a cellular pattern (general IHC practice); HPA reports cytoplasmic and nuclear tissue staining (HPA: tissue IHC profile). | Review blocking, washes, antibody concentration and development time alongside a control lacking primary antibody (general IHC practice). |
| Nuclear staining appears alongside cytoplasmic staining. | Nuclear expression is included in the HPA tissue profile, and UniProt lists the nuclear envelope as a location (HPA: tissue IHC; UniProt Q14203 subcellular location). | Score nuclear and cytoplasmic compartments separately; judge each against tissue morphology and controls (general IHC practice). |
| IF/ICC question: what pattern should be visible? | HPA supports microtubules as the main location and the cytokinetic bridge as an additional location; it marks spindle, primary-cilium and basal-body locations uncertain (HPA: subcellular ICC-IF). | Use the supported microtubule pattern as the primary localization check and interpret uncertain locations cautiously (HPA: subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing DCTN1 staining
A02175-2 · IHCIHC analysis of DCTN1/p150-glued using anti-DCTN1/p150-glued antibody (A02175-2). DCTN1/p150-glued was detected in paraffin-embedded section
A02175-2 · IHCIHC analysis of DCTN1/p150-glued using anti-DCTN1/p150-glued antibody (A02175-2). DCTN1/p150-glued was detected in paraffin-embedded section
A02175-2 · IFIF analysis of DCTN1/p150-glued using anti-DCTN1/p150-glued antibody (A02175-2). DCTN1/p150-glued was detected in immunocytochemical section
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000204843-DCTN1/tissue/cerebral+cortex#imid_21249827
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000204843-DCTN1/tissue/colon#imid_21249753
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000204843-DCTN1/tissue/liver#imid_21249754
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000204843-DCTN1/tissue/kidney#imid_21249739
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA071875; green = DCTN1, blue = nucleus, red = microtubules.
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA071875; green = DCTN1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DCTN1 IHC & IF
🧪Run fallopian tube first and expect staining in glandular cells (HPA: High in fallopian tube glandular cells); use lymph node germinal center cells as the negative tissue comparator (HPA: Not detected). On the positive slide, cells without visible staining can serve as internal background comparators, but the supplied HPA rows do not establish any cell type there as DCTN1-negative.
Positive control tissue: Fallopian tube (Glandular cells, HPA High)
Negative control tissue: Lymph node (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show DCTN1 in HEL, Rh30, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), Sperm, with annotated localisation: Microtubules (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a rabbit IgG isotype control matched to the primary antibody's clonality, and DCTN1 knockout material as a biological negative (caption: rabbit primary; standard IHC control practice). Quench endogenous peroxidase and check for endogenous biotin in the positive tissue when using the documented SABC/DAB detection (caption: SABC/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A02175-2 paraffin-section caption does not report a fixative (caption: fixative not stated). That caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required for DCTN1 (caption: EDTA retrieval). Frozen sections cannot be ranked as easier from the supplied evidence; IF offers a supported microtubule localization readout in its separate guide, while tissue-specific artefacts are unreported (HPA subcellular: microtubules supported).
HPA tissue IHC evidence for DCTN1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced DCTN1 IHC Tips
Use the IHC-validated antibody’s paraffin-section example as a starting point, then judge staining against DCTN1’s documented distribution and appropriate controls.
Which retrieval conditions should I start with for DCTN1 paraffin-section IHC?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A02175-2). The selected paraffin-section example used this condition before staining human ovarian adenocarcinoma tissue with the catalog antibody at 2 µg/ml overnight at 4°C (datasheet A02175-2). If staining is weak, compare a carefully controlled retrieval time series while keeping antibody concentration and detection constant; excessive heating can damage tissue morphology (standard IHC practice). Include a known positive section in every comparison, and judge improvement by intracellular signal with preserved cellular detail rather than by stronger staining alone (HPA: cytoplasmic and nuclear expression in most tissues; standard IHC practice).
How should I troubleshoot weak staining when fixation details are missing?
Target-specific DCTN1 sensitivity to fixation is unknown from the supplied evidence (datasheet A02175-2: fixative not stated). The selected image describes a paraffin-embedded section, so its retrieval and staining conditions cannot establish how a particular fixative affects epitope preservation (datasheet A02175-2). Record the actual fixative and fixation duration for each specimen, then compare sections processed and stained together with the same EDTA pH 8.0 retrieval (standard IHC practice; datasheet A02175-2). If signal varies, assess tissue morphology and positive-control staining before changing antibody concentration, because processing differences can confound a direct comparison (standard IHC practice).
What DCTN1 staining pattern is plausible in tissue sections?
Expect mainly intracellular staining: DCTN1 is reported in cytoplasm, cytoskeletal structures, centrosomes, spindle, cell cortex and nuclear envelope (UniProt Q14203 subcellular location). HPA reports cytoplasmic and nuclear staining across most tissues, while its cell-imaging evidence supports microtubule localisation (HPA tissue IHC; HPA subcellular). In chromogenic sections, use the counterstain to distinguish cytoplasmic signal from apparent nuclear overlap, and inspect multiple intact cells before assigning a compartment (standard IHC practice). A narrow rim around a nucleus could fit the reported nuclear-envelope location, but diffuse nuclear DAB should be checked against controls before receiving that interpretation (UniProt Q14203 subcellular location; standard IHC practice).
Could isoforms or epitope access explain uneven DCTN1 staining?
DCTN1 has 6 listed isoforms, including p150 and p135, and a CAP-Gly domain at residues 48–90 (UniProt Q14203 isoforms and domains). The supplied caption names p150-glued but does not map the antibody epitope or establish which isoforms it detects in tissue (datasheet A02175-2). Do not assign a cell-specific staining difference to isoform expression without epitope information and independent supporting evidence (UniProt Q14203 isoforms; standard IHC practice). Compare retrieval conditions on matched sections and keep detection settings fixed; altered epitope access can change apparent staining without demonstrating a biological change (standard IHC practice).
How should I adapt this DCTN1 question to multiplex IF?
For multiplex IF, pair DCTN1 with a marker identifying the cell population under study, such as the glandular cells reported as strongly stained in fallopian tube or gallbladder (HPA tissue IHC). Choose spectrally separated fluorophores and assess an unstained section for tissue autofluorescence before assigning channels; a far-red channel can be useful when shorter-wavelength background is high (standard IF practice). DCTN1 is intracellular and has no transmembrane segment, so permeabilise after fixation when access to a cytoplasmic epitope is required, adjusting conditions to preserve cell structure (UniProt Q14203 topology and subcellular location; standard IF practice). Include single-label controls to evaluate channel bleed-through and interpret puncta against cellular morphology (standard IF practice).
What should I check when DCTN1 DAB staining is widespread?
Widespread staining is not automatically background: HPA reports cytoplasmic and nuclear expression in most tissues, with high staining in glandular cells of fallopian tube and gallbladder (HPA tissue IHC). Compare intact-cell localisation with a no-primary control and inspect tissue edges, folds and damaged areas for nonspecific deposition (standard IHC practice). The selected example used 10% goat serum blocking, biotinylated secondary antibody, streptavidin-biotin detection and DAB, so evaluate background across that detection chain if using the same workflow (datasheet A02175-2). Apply an endogenous peroxidase block and appropriate detection controls when troubleshooting DAB signal; these are general chromogenic IHC measures (standard IHC practice).
How can I quantify DCTN1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because HPA reports both cytoplasmic and nuclear tissue staining (HPA tissue IHC). For cell-level comparisons, record the percentage of positive cells and an H-score from intensity categories; use identical thresholds and blinded scoring across groups (standard IHC practice). If measuring spatial abundance, report positive-cell density per mm² of viable tissue and normalise to the relevant cell population or tissue area, rather than comparing raw counts from unequal sections (standard IHC practice). Keep retrieval, DAB development and imaging conditions consistent, and report cytoplasmic and nuclear scores separately when both are assessed (standard IHC practice; HPA tissue IHC).
How do I distinguish genuine DCTN1 signal from artefact?
A plausible positive signal lies within intact cells and agrees with DCTN1’s reported cytoplasmic, cytoskeletal or nuclear-envelope locations; HPA also reports cytoplasmic and nuclear tissue staining (UniProt Q14203 subcellular location; HPA tissue IHC). Check whether the stained cells match the population being scored: HPA reports high staining in fallopian-tube and gallbladder glandular cells but no detection in lymph-node germinal-center cells (HPA tissue IHC). Treat staining restricted to section edges, folds or necrotic areas as suspect and compare it with adjacent intact tissue (standard IHC practice). A no-primary control helps identify detection background, including endogenous enzyme signal in a DAB workflow (standard IHC practice).
Boster reagents
Best DCTN1 / Dynactin subunit 1 IHC Antibodies
A02175-2 has human paraffin-section IHC images (IHC captions) and a CACO-2 cell IF image (IF caption); catalog reactivity covers human, mouse and rat (catalog: reactivity).
Real IHC data
Anti-DCTN1/p150-glued Antibody ®
Cat # A02175-2
A02175-2 has IHC images from human paraffin-embedded ovarian adenocarcinoma, placenta and breast cancer sections (IHC captions). The same SKU has an IF image from CACO-2 cells (IF caption) and lists IHC, ICC and IF applications (catalog: applications).
Which to pick: For paraffin-section IHC, choose A02175-2: its human tissue captions use EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (IHC captions); the fixative is unreported (IHC captions). For IF/ICC, A02175-2 has a CACO-2 cell IF image at 5 μg/ml (IF caption). For mouse or rat samples, A02175-2 lists those species as reactive (catalog: reactivity), while the supplied IHC and IF images show human samples (IHC and IF captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q14203 (DCTN1_HUMAN, Dynactin subunit 1).
- Human Protein Atlas. DCTN1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. DCTN1 subcellular location (ICC-IF): Mainly localized to the microtubules. In addition localized to the cytokinetic bridge, mitotic spindle, primary cilium, basal body, acrosome and perinuclear theca..
- Human Protein Atlas. DCTN1 antibody validation summary (4 antibodies).
- Dynactin Deficiency in the CNS of Humans with Sporadic ALS and Mice with Genetically Determined Motor Neuron Degeneration. Neurochemical research 2013 — PMC3898179.
- Dysregulation of stress granule dynamics by DCTN1 deficiency exacerbates TDP-43 pathology in Drosophila models of ALS/FTD. Acta neuropathologica communications 2024 — PMC10840176.
- Human Mutant Dynactin Subunit 1 Causes Profound Motor Neuron Disease Consistent with Possible Mechanisms Involving Axonopathy, Mitochondriopathy, Protein Nitration, and T-Cell-Mediated Cytolysis. Biomolecules 2025 — PMC12730482.
- DCTN1 Binds to TDP-43 and Regulates TDP-43 Aggregation. International journal of molecular sciences 2021 — PMC8070438.
- PubMed PMID:9799602 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DCTN1 (Dynactin subunit 1). Expected localisation Cytoplasmic and nuclear staining in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02175-2). Validated IHC antibody: A02175-2 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Fallopian tube (Glandular cells); negative Lymph node. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human ovarian adenocarcinoma tissue; fixative not specified (datasheet A02175-2); 2. Fixation: Image fixative and duration unreported (datasheet A02175-2); 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 A02175-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02175-2); 8. Primary antibody: Rabbit anti-DCTN1, 2-5μg/ml (datasheet A02175-2); 9. Primary incubation: Overnight at 4 °C (datasheet A02175-2). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Germinal center cells may lack detectable staining (HPA tissue IHC); Isoform / epitope: 6 isoforms; antibody epitope coverage is unspecified (UniProt); Regulation: No expression regulator specified (UniProt). Evidence: UniProt Q14203, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the microtubules. In addition localized to the…, Datasheet A02175-2, PMC3898179, PMC12730482. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DCTN1/immunohistochemistry-ihc-guide.