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DCTN1 / Dynactin subunit 1 · IHC design guide
Design Immunohistochemistry for DCTN1
Plan chromogenic DCTN1 IHC on paraffin sections around the cytoplasmic and nuclear staining seen in most tissues (HPA tissue IHC). This guide covers fixation consistency, cell-specific interpretation (HPA tissue IHC), and the catalog antibody’s 2–5 μg/mL IHC concentration (datasheet A02175-2).
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 tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic and nuclear staining in most tissues (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 paraffin 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; epitope coverage is unspecified (UniProt) |
Section 1
Recommended DCTN1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A02175-2) is accompanied by a published DCTN1 IHC protocol for human CNS tissue (PMC3898179).
| 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 retrieval for the catalog antibody (datasheet A02175-2); the published DCTN1 method does not specify retrieval for DCTN1 (PMC3898179).
Section 2
What Is the Expected DCTN1 Staining Pattern?
DCTN1 is a non-transmembrane protein associated with cytoplasm, cytoskeleton, centrosomes and the nuclear envelope (UniProt Q14203 topology and subcellular location). In paraffin-section IHC, expect cytoplasmic and sometimes nuclear staining across many tissues, including strong staining in fallopian tube and gallbladder glandular cells (HPA: tissue IHC). HPA rates its tissue staining Approved, with medium consistency against RNA data and external verification pending (HPA: reliability).
| Strong cytoplasmic staining in fallopian tube or gallbladder glandular cells. | This matches the reported high staining in those cells (HPA: High in fallopian tube and gallbladder glandular cells). Compare signal within the glandular cells and surrounding tissue; the reported level describes those cell populations, not every cell in the section (HPA: tissue IHC). |
| Cytoplasmic staining with some nuclear or perinuclear staining. | HPA reports cytoplasmic and nuclear expression in most tissues (HPA: tissue IHC profile); UniProt also places DCTN1 at the nuclear envelope (UniProt Q14203 subcellular location). Do not classify every nuclear signal as artefact on compartment alone. Score the cells and tissue against the HPA pattern. |
| Signal confined to the cell surface, extracellular material, or an unrelated compartment. | That distribution does not fit the reported cytoplasmic, cytoskeletal and nuclear-envelope locations or the absence of a transmembrane segment (UniProt Q14203 subcellular location and topology). Treat an isolated wrong-compartment pattern as suspect; review controls and detection background before calling it DCTN1. |
| Strong staining in cells reported as not detected, or broad colour outside cell boundaries. | Germinal-center cells in lymph node and red-pulp cells in spleen are listed as not detected (HPA: tissue IHC). Signal there warrants a cross-reactivity or endogenous-detection check (standard IHC practice). Diffuse colour across cells and matrix likewise suggests background rather than a resolved cellular pattern (standard IHC practice). |
| No detectable signal in fallopian tube or gallbladder glandular cells. | These are reported high-staining populations, so an absent result calls for a run-level check before interpreting the sample as DCTN1-negative (HPA: tissue IHC; standard IHC practice). Confirm section integrity, the antibody's IHC-P instructions and detection controls; no target-specific fixation sensitivity is established by the supplied sources. |
💡Expected DCTN1 appearanceA convincing positive paraffin-section result is predominantly cellular cytoplasmic staining, potentially with nuclear signal, and strong staining in fallopian tube or gallbladder glandular cells; isolated surface or extracellular colour is suspect (HPA: tissue IHC; UniProt Q14203 subcellular location and topology).
| Compartment and tissue context | UniProt places DCTN1 in several cytoskeletal structures and at the nuclear envelope, while HPA describes cytoplasmic and nuclear tissue staining (UniProt Q14203 subcellular location; HPA: tissue IHC profile). Assess the compartment together with the stained cell population; a single subcellular feature is not an IHC-P validation criterion. |
| Choice of positive and low-signal comparators | Fallopian tube and gallbladder glandular cells are reported High; hippocampal neuronal cells, esophageal squamous cells and bone-marrow hematopoietic cells are reported Low (HPA: tissue IHC). Use a High population to check whether the run detects the expected pattern. A Low population alone gives a weaker absence-of-signal test. |
| Antibody evidence and isoforms | HPA lists four rabbit polyclonal antibodies as IHC Approved, while its overall tissue profile has medium RNA-staining consistency and awaits external verification (HPA: antibodies and reliability). UniProt lists six DCTN1 isoforms (UniProt Q14203 isoforms). The supplied evidence gives no antibody epitope map, so isoform coverage cannot be inferred. |
| IF/ICC Q&A: should the IHC section look like an IF microtubule image? | No direct image-to-section match is established: HPA supports microtubule localization in ICC-IF, and also reports additional structures with differing confidence (HPA: subcellular ICC-IF). For this paraffin-section IHC guide, judge the tissue's observed cell and compartment pattern using HPA tissue IHC; follow the separate IF/ICC guide for imaging. |
| Situation | Likely cause | Next action |
|---|---|---|
| High-staining glandular cells show no signal. | A failed IHC run or unsuitable assay conditions are possible; the supplied sources do not identify a DCTN1-specific fixation effect (HPA: High in fallopian tube and gallbladder glandular cells; standard IHC practice). | Check the catalog antibody's IHC-P instructions, section quality and a known-positive tissue on the same run; inspect the detection control before assigning a negative result (standard IHC practice). |
| Colour covers most of the section without clear cell boundaries. | Nonspecific binding or detection background can obscure a cellular IHC pattern (standard IHC practice). HPA's broad tissue expression alone does not explain uniform colour in cells and extracellular space (HPA: tissue IHC profile). | Inspect the negative-reagent control, blocking and washes; then adjust antibody or detection conditions according to the validated IHC-P workflow (standard IHC practice). |
| Unexpected strong staining appears in lymph-node germinal-center cells or splenic red-pulp cells. | Those populations are reported Not detected (HPA: tissue IHC). Cross-reactivity or endogenous detection activity is possible, but the unexpected signal alone does not distinguish them (standard IHC practice). | Compare a reagent control that omits primary antibody with the full stain; review blocking and cell morphology before attributing the colour to DCTN1 (standard IHC practice). |
| Staining appears only on membranes or in extracellular material. | The pattern conflicts with DCTN1's reported intracellular locations and lack of a transmembrane segment (UniProt Q14203 subcellular location and topology). Section-edge or detection artefact should be considered (standard IHC practice). | Check whether the same pattern appears in controls, and repeat assessment in an HPA High glandular population with clear cellular morphology (HPA: tissue IHC; standard IHC practice). |
| Nuclear signal seems stronger than expected. | Nuclear tissue staining is within the HPA profile, and UniProt reports nuclear-envelope localization; intensity alone does not establish its origin (HPA: tissue IHC profile; UniProt Q14203 subcellular location). | Compare nuclear and cytoplasmic staining cell by cell, examine the negative-reagent control, and report the observed compartments without forcing a cytoplasm-only score (standard IHC practice; HPA: tissue IHC profile). |
| A low-staining comparator looks negative while the positive control works. | HPA reports Low staining in hippocampal neuronal, esophageal squamous and bone-marrow hematopoietic cells (HPA: tissue IHC). A weak population may fall below visibility under the selected IHC conditions (standard IHC practice). | Keep the successful High glandular-cell control as the run check, record the low comparator's observed intensity, and avoid declaring the antibody failed from that comparator alone (HPA: tissue IHC; standard IHC practice). |
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 confirm staining in its glandular cells (HPA: High in fallopian tube glandular cells). Use lymph node germinal center cells as the negative tissue comparator (HPA: Not detected in lymph node germinal center cells); on the positive slide, assess other cells for background relative to glandular cells without treating them as validated DCTN1-negative cells.
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 and a concentration-matched rabbit IgG isotype control appropriate to the primary antibody (selected-SKU caption: rabbit anti-DCTN1 antibody). A DCTN1 knockout specimen, if available, provides a biological negative; quench endogenous peroxidase and check for endogenous biotin because the caption uses a biotin-based system with DAB (selected-SKU caption: biotinylated secondary, SABC and DAB).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the fixative is unreported in the selected-SKU paraffin-section caption. That caption uses heat retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required under other conditions (selected-SKU caption: EDTA heat retrieval). The supplied evidence does not establish whether frozen sections or IF are easier, and reports no tissue-specific artefact for the selected fallopian tube control.
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
Troubleshoot DCTN1 staining in paraffin sections using the catalog antibody’s tissue protocol and documented localisation as reference points.
What retrieval should I try first if DCTN1 staining is weak?
Start with heat-mediated EDTA retrieval at pH 8.0 for paraffin sections (datasheet A02175-2). Keep retrieval conditions consistent across comparison sections, then check whether the catalog antibody detects cytoplasmic signal in glandular cells (datasheet A02175-2; HPA tissue IHC). If signal remains weak, adjust heating or cooling within your laboratory’s established retrieval workflow while holding antibody concentration constant (standard IHC practice). The documented tissue image used 2 µg/mL primary antibody overnight at 4°C, providing a starting concentration and incubation for this SKU (datasheet A02175-2). Assess morphology and background alongside signal, because overly harsh retrieval can impair interpretation (standard IHC practice).
Could fixation explain faint or uneven DCTN1 staining?
Target-specific fixation sensitivity is unknown: the selected tissue-IHC caption identifies paraffin sections but does not state a fixative (datasheet A02175-2). Record the actual fixative and processing history for each specimen before comparing staining intensity across cases (standard IHC practice). When troubleshooting, compare sections with similar processing histories and repeat the same pH 8.0 EDTA retrieval and 2 µg/mL primary incubation (datasheet A02175-2; standard IHC practice). Inspect tissue edges and poorly preserved regions separately, since uneven preservation can complicate chromogenic scoring (standard IHC practice). Do not assign a DCTN1-specific fixation effect from tissue expression or protein features alone (HPA tissue IHC; UniProt Q14203; datasheet A02175-2).
Where should convincing DCTN1 signal appear in tissue sections?
Expect predominantly cytoplasmic staining, while allowing for nuclear-associated signal reported across tissues (HPA tissue IHC). DCTN1 is also annotated at microtubules, centrosomes, spindles and the nuclear envelope, so cellular context matters when assessing focal patterns (UniProt Q14203 subcellular). High staining in fallopian-tube and gallbladder glandular cells offers documented tissue context for comparison (HPA tissue IHC). Avoid calling a small brown focus a centrosome from chromogenic staining alone; assess cellular morphology and, where needed, a separately validated localisation method (UniProt Q14203 subcellular; standard IHC practice). Compare signal with a no-primary control and intact neighboring tissue before assigning a subcellular pattern (standard IHC practice).
Can this IHC assay distinguish DCTN1 isoforms or phosphorylation states?
DCTN1 has 6 annotated isoforms, including p150 and p135, and a CAP-Gly domain at residues 48–90 (UniProt Q14203). The supplied tissue-IHC caption does not map the antibody epitope, so staining alone cannot establish which isoforms it recognizes (datasheet A02175-2; UniProt Q14203). Documented modifications include phosphorylation at Thr-108, Thr-145–147 and Ser-179, but no phosphorylation-specific reactivity is supplied for this antibody (UniProt Q14203; datasheet A02175-2). Interpret intensity as antibody-detected DCTN1 signal, and request epitope-mapping or isoform-specific validation before making finer claims (standard IHC practice). Keep the same retrieval and detection conditions when comparing specimens (datasheet A02175-2; standard IHC practice).
How should I investigate DCTN1 localisation by multiplex IF?
Use this as an IF investigation separate from the documented chromogenic paraffin-section assay; the selected caption supplies no IF conditions (datasheet A02175-2). Pair DCTN1 with a validated marker identifying the cell population being assessed, then examine overlap within individual cells rather than adjacent cells (standard IF practice). Choose spectrally separated fluorophores and assign the weaker channel away from the tissue’s strongest measured autofluorescence (standard IF practice). DCTN1 has no transmembrane segment and is annotated in intracellular compartments, so use permeabilisation compatible with access to its cytoplasmic epitope (UniProt Q14203 topology and subcellular; standard IF practice). Include single-stain and no-primary controls to check bleed-through and background (standard IF practice).
How can I reduce diffuse brown background without losing DCTN1 signal?
First compare the stained section with a no-primary control to identify secondary-reagent or detection background (standard IHC practice). The documented assay blocked with 10% goat serum and used biotinylated goat anti-rabbit secondary for 30 minutes at 37°C (datasheet A02175-2). Check endogenous peroxidase blocking and the biotin-based detection controls before attributing diffuse DAB deposition to DCTN1 (datasheet A02175-2; standard IHC practice). If background persists, optimize washing and primary concentration while retaining a section processed at the documented 2 µg/mL condition for comparison (datasheet A02175-2; standard IHC practice). Judge improvement by preserved cell-associated signal and reduced staining in the no-primary control (standard IHC practice).
How should I score DCTN1 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because DCTN1 has documented cytoplasmic and nuclear-associated patterns (HPA tissue IHC; UniProt Q14203 subcellular). Record the percentage of positive cells and staining intensity on a 0–3 scale, then calculate an H-score from those values (standard IHC practice). For spatial questions, report positive-cell density per mm² of viable assessed tissue and retain the region definitions (standard IHC practice). Normalize comparisons to the same eligible cell population or viable tissue area, with matched retrieval, detection and imaging conditions (standard IHC practice). Exclude necrotic or damaged regions by a prespecified rule and report exclusions alongside scores (standard IHC practice).
Which staining patterns suggest a true DCTN1 signal rather than artefact?
Favor reproducible cell-associated cytoplasmic staining that fits morphology; nuclear-associated staining can also be plausible for DCTN1 (HPA tissue IHC; UniProt Q14203 subcellular). Glandular cells in fallopian tube and gallbladder show high reported staining, whereas lymph-node germinal-center cells and splenic red-pulp cells were not detected (HPA tissue IHC). Treat isolated edge staining, necrotic-area staining or color persisting in a no-primary control as possible artefact (standard IHC practice). Check peroxidase and biotin-based detection controls when DAB deposition lacks a coherent cellular pattern (datasheet A02175-2; standard IHC practice). HPA rates tissue evidence as approved with medium staining–RNA consistency pending external verification, so interpret unexpected patterns cautiously (HPA tissue IHC).
Boster reagents
Best DCTN1 / Dynactin subunit 1 IHC Antibodies
A02175-2 has paraffin-section IHC images from human ovarian adenocarcinoma, placenta and breast cancer, plus an IF image from CACO-2 cells (catalog image captions).
Real IHC data
Anti-DCTN1/p150-glued Antibody ®
Cat # A02175-2
A02175-2 is listed for IHC and has images from paraffin sections of human ovarian adenocarcinoma, placenta and breast cancer (catalog applications; IHC image captions). The same SKU is listed for IF/ICC and has an IF image from CACO-2 cells (catalog applications; IF image caption).
Which to pick: Choose A02175-2 for human tissue IHC because its own images document paraffin sections stained at 2 μg/mL; the fixative is unreported (A02175-2 IHC image captions). Choose the same SKU for IF/ICC because its IF image documents CACO-2 cells stained at 5 μg/mL (A02175-2 IF image caption). Human, mouse and rat reactivity is listed, but the supplied IHC and IF images show human samples only, so verify staining in mouse or rat samples before use (catalog reactivity; A02175-2 image 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 tissue staining (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 paraffin sections (standard IHC practice; not target-specific); Caveat: Germinal center cells may lack detectable staining (HPA tissue IHC); Isoform / epitope: 6 isoforms; 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. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DCTN1/immunohistochemistry-ihc-guide.