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DOCK3 / Dedicator of cytokinesis protein 3 · IHC design guide
Design Immunohistochemistry for DOCK3
Plan DOCK3 chromogenic IHC in paraffin sections using the IHC-validated antibody and cerebral cortex neuronal cells as a positive reference (HPA tissue IHC). The guide covers fixation consistency, staining controls and interpretation of the expected cytoplasmic pattern (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DOCK3 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 CNS (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in CNS neuronal cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A08904) | |
| Positive control | Cerebral cortex+4 more · see all | |
| Negative control | Adrenal gland+4 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). Selected-image fixative and duration unreported (datasheet A08904); verify before use. | |
| Caveat | Staining has medium concordance with RNA data (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | No listed isoforms or processing; epitope site unspecified (UniProt) |
Section 1
Recommended DOCK3 IHC & IF Protocols
The catalog antibody has a paraffin-section IHC protocol (datasheet A08904). One published Dock3 IHC protocol uses frozen optic-nerve sections (PMC4454328).
| Sample | Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet A08904) |
| Fixation | Image fixative and duration unreported (datasheet A08904); 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 A08904); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08904) |
| Primary antibody | Rabbit anti-DOCK3, 2-5 μg/ml (datasheet A08904) |
| Primary incubation | Overnight at 4 °C (datasheet A08904) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A08904) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DOCK3-positive staining in neuronal cells of cerebral cortex (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in CNS. No signal in the no-primary control. |
💡Decision noteFor paraffin sections, start with heat-mediated EDTA pH 8.0 retrieval (datasheet A08904). The published frozen-section method is a separate starting point for cryosections (PMC4454328).
Section 2
What Is the Expected DOCK3 Staining Pattern?
DOCK3 is primarily cytoplasmic and has no transmembrane segment (UniProt Q8IZD9). In paraffin sections, expect the clearest staining in cerebral cortex neuronal cells, with weaker or variable staining elsewhere (HPA: High in cerebral cortex neuronal cells; tissue IHC reliability Enhanced). HPA describes cytoplasmic expression in the CNS, but its tissue profile has medium consistency with RNA data and awaits external verification (HPA: tissue IHC profile and reliability description).
| Neuronal cytoplasm stains strongly in cerebral cortex; nearby cells and structures vary. | This fits the strongest listed tissue result (HPA: High in cerebral cortex neuronal cells). Neuropil staining can also fit normal brain distribution (UniProt Q8IZD9: normal brain tissue specificity). Judge the cell type and compartment together; a brown signal alone does not identify DOCK3 (general IHC practice). |
| Predominantly nuclear or sharply outlined membrane staining replaces the expected cytoplasmic pattern. | A dominant nuclear pattern conflicts with the reported cytoplasmic location (UniProt Q8IZD9; HPA: cytoplasmic expression in CNS). DOCK3 can become membrane associated through PSEN1 interaction, so limited membrane proximity is plausible; a dominant membrane outline needs control-based review (UniProt Q8IZD9: PSEN1 interaction; general IHC practice). |
| Strong staining appears in a cell population listed as undetected by HPA. | For example, strong adrenal glandular-cell staining conflicts with that listed result (HPA: Not detected in adrenal glandular cells). Consider nonspecific antibody binding or endogenous chromogenic activity, then compare a no-primary control (general IHC practice). Do not treat every non-neuronal cell as suspect: several non-neuronal populations have Medium staining (HPA: skin, stomach, testis). |
| Pale brown color spreads across cells and tissue without a discernible cytoplasmic pattern. | Diffuse color is difficult to score as DOCK3 because the expected pattern is compartmental (HPA: cytoplasmic expression in CNS; UniProt Q8IZD9: cytoplasm). Check the no-primary control and detection reagents, then optimize blocking, washing and antibody concentration as general IHC practice; no DOCK3-specific cause follows from this appearance alone. |
| Cerebral cortex neuronal cells show no convincing signal. | This is unexpected relative to the strongest listed tissue result (HPA: High in cerebral cortex neuronal cells). First verify tissue identity and the presence of evaluable neurons; then check the IHC antibody, retrieval and detection workflow (general IHC practice). HPA's Enhanced rating still has medium RNA–staining consistency and pending external verification (HPA: reliability description). |
💡Expected DOCK3 appearanceCall a positive result when cerebral cortex neuronal cells show clear, predominantly cytoplasmic staining, potentially extending into neuropil; strong nuclear staining or uniform color across HPA-undetected populations warrants control review (HPA: High in cerebral cortex neuronal cells; UniProt Q8IZD9: normal brain tissue specificity; HPA: negative tissue list).
| Tissue and cell selection | Cerebral cortex neuronal cells offer the strongest listed IHC signal; caudate neurons are Medium and hippocampal neurons Low (HPA: tissue IHC). A weak hippocampal result alone is less informative than a failed cerebral cortex control. HPA also lists Medium staining in skin keratinocytes, stomach glandular cells and testis Leydig cells (HPA: tissue IHC). |
| Compartment and membrane association | DOCK3 is cytoplasmic and lacks a transmembrane segment (UniProt Q8IZD9). Its PSEN1 interaction can mediate membrane association, so some signal near a membrane can be biologically plausible (UniProt Q8IZD9: subunit and subcellular location). This does not establish that a dominant membrane outline is the expected paraffin-section pattern. |
| Antibody evidence | HPA037543 has Enhanced IHC validation, meaning its pattern was reproduced by independent antibodies or orthogonal data (HPA: antibody validation summary). HPA037544 has an Enhanced ICC rating but no supplied IHC rating (HPA: antibody validation summary). Interpret either rating alongside the tissue profile's medium RNA–staining consistency and pending external verification (HPA: reliability description). |
| Processing and epitope limits | UniProt reports one 1–2030 chain, no signal peptide, no propeptide and no listed isoforms (UniProt Q8IZD9). Those annotations do not identify the antibody epitope or establish an antigen-retrieval condition. No DOCK3-specific fixation sensitivity is supplied; select and document retrieval using the IHC antibody's validated procedure (general IHC practice). |
| IF/ICC question: what pattern is expected? | Predominantly cytosolic fluorescence is the HPA ICC-IF result, with an Enhanced cytosol location (HPA: subcellular summary). HPA lists ICC-IF images in A-431, U-251MG and U2OS, and Enhanced ICC ratings for HPA037543 and HPA037544 (HPA: subcellular and antibody records). Those ICC observations do not establish an IHC retrieval or chromogenic protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Cerebral cortex has no neuronal staining. | The strongest listed positive population is missing (HPA: High in cerebral cortex neuronal cells); the specific technical cause cannot be inferred from the image alone. | Confirm evaluable neurons and tissue identity, then check primary-antibody use, retrieval and chromogenic detection against the IHC procedure; include a working positive control (general IHC practice). |
| Nuclei stain more strongly than cytoplasm. | Dominant nuclear staining conflicts with the reported cytoplasmic location (UniProt Q8IZD9; HPA: CNS cytoplasmic profile). | Compare with a no-primary control and inspect counterstain and chromogen separately; reassess antibody specificity before scoring nuclear signal as DOCK3 (general IHC practice). |
| Strong signal appears in an HPA-undetected population. | An unexpected cell pattern may reflect nonspecific binding or endogenous detection activity (general IHC practice); HPA lists adrenal glandular cells as Not detected (HPA: tissue IHC). | Run a no-primary control, check detection chemistry and blocking, and compare the suspect cells with cerebral cortex neurons on the same staining run (general IHC practice). |
| Brown haze obscures cell boundaries. | Diffuse background can arise from the staining workflow; its appearance alone does not identify a DOCK3-specific mechanism (general IHC practice). | Inspect the no-primary control, improve washing or blocking if indicated, and titrate the primary antibody within its validated IHC procedure (general IHC practice). |
| Hippocampus is weak while cerebral cortex stains clearly. | The difference matches the listed levels: Low in hippocampal neurons and High in cerebral cortex neuronal cells (HPA: tissue IHC). | Score each tissue against its own expected level and compartment; retain cerebral cortex as the stronger positive comparator (HPA: tissue IHC; general IHC practice). |
| A strong membrane rim is scored as positive. | PSEN1-mediated membrane association is possible, but DOCK3 is primarily cytoplasmic and has no transmembrane segment (UniProt Q8IZD9). | Review the cytoplasmic signal and control sections before accepting the rim; record membrane enrichment separately from the expected cytoplasmic pattern (UniProt Q8IZD9; general IHC practice). |
Experiment evidence
IHC & IF images showing DOCK3 staining
A08904 · IHCIHC analysis of MOCA/DOCK3 using anti-MOCA/DOCK3 antibody (A08904). MOCA/DOCK3 was detected in a paraffin-embedded section of human colon ca
A08904 · IHCIHC analysis of MOCA/DOCK3 using anti-MOCA/DOCK3 antibody (A08904). MOCA/DOCK3 was detected in a paraffin-embedded section of human lung can
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000088538-DOCK3/tissue/cerebral+cortex#imid_1337
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000088538-DOCK3/tissue/cerebral+cortex#imid_20712424
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000088538-DOCK3/tissue/colon#imid_20712334
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000088538-DOCK3/tissue/liver#imid_20712351
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037543; green = DOCK3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037543; green = DOCK3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DOCK3 IHC & IF
🧪Run cerebral cortex first: neuronal cells should stain strongly (HPA: High in cerebral cortex neuronal cells). Use adrenal gland glandular cells as the negative tissue (HPA: Not detected); on the cortex slide, adjacent non-neuronal cells should show little signal relative to neuronal cells, with any staining assessed against the negative controls (HPA: High in neuronal cells; standard IHC practice).
Positive control tissue: Cerebral cortex (Neuronal cells, HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show DOCK3 in A-431, U-251MG, U2OS, with annotated localisation: Cytosol (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; rabbit IgG matched to the primary antibody’s clonality and concentration; and a DOCK3 knockout specimen where available as a biological negative (A08904 caption: rabbit primary; standard IHC practice). Block endogenous peroxidase and assess pigment-related background in cerebral cortex before interpreting DAB staining (HPA: cerebral cortex; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected A08904 tissue-IHC caption does not state the fixative (A08904 caption). The caption uses heat-mediated retrieval in EDTA at pH 8.0 for a paraffin section; it does not establish whether DOCK3 staining depends on retrieval, and the evidence does not establish that frozen sections or IF are easier (A08904 caption). In cerebral cortex, neuronal pigment can complicate interpretation of chromogenic staining, so compare the signal with the no-primary control (HPA: High in cerebral cortex neuronal cells; standard IHC practice).
HPA tissue IHC evidence for DOCK3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Skin | Keratinocytes | Medium | Protein (IHC) | HPA → |
| Stomach | Glandular cells | Medium | Protein (IHC) | HPA → |
| Testis | Leydig cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced DOCK3 IHC Tips
Troubleshoot DOCK3 staining in paraffin sections using the catalog antibody’s tissue example, expected cytoplasmic pattern, and appropriate controls.
What retrieval should I try first for weak DOCK3 staining in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA pH 8.0 (datasheet A08904). The catalog antibody’s paraffin-section example used that retrieval before an overnight incubation with 2 μg/ml primary antibody at 4°C (datasheet A08904). If staining remains weak, check heating consistency, section adhesion, and reagent performance before comparing a second retrieval condition on matched sections (standard IHC practice). Keep detection and exposure conditions identical across that comparison, and include a known positive section so that a retrieval change can be judged against the expected cytoplasmic signal (HPA tissue IHC: cytoplasmic expression in CNS; standard IHC practice).
Could fixation explain weak or patchy DOCK3 staining?
Target-specific fixation sensitivity is unknown: the A08904 paraffin-section caption does not state a fixative (datasheet A08904). Record the fixative and fixation duration for each specimen, then compare similarly processed sections when investigating patchy staining (standard IHC practice). If fixation differs between samples, assess tissue morphology and a separate process-control stain before attributing a DOCK3 difference to biology (standard IHC practice). Keep retrieval at EDTA pH 8.0 and the primary incubation consistent while making that comparison (datasheet A08904; standard IHC practice). Neither the reported tissue pattern nor DOCK3 topology establishes how this antibody responds to a particular fixative (HPA tissue IHC; UniProt Q8IZD9 topology).
Where should I expect DOCK3 signal, and how should I assess membrane staining?
Expect predominantly cytoplasmic staining, consistent with the reported CNS tissue profile and cytosolic cell images (HPA tissue IHC; HPA subcellular: cytosol). In normal brain, DOCK3 is reported in neuropil and occasionally in pyramidal cells, so assess those structures separately when they are present (UniProt Q8IZD9 tissue specificity). DOCK3 has no transmembrane segment, although interaction with PSEN1 can make it membrane associated; a focal membrane-adjacent pattern therefore needs corroboration rather than automatic rejection (UniProt Q8IZD9 topology and subunit). Compare the pattern with morphology and a matched negative control, especially where dense processes make cytoplasmic boundaries difficult to resolve (standard IHC practice).
How can I investigate discordant staining when the antibody epitope is unspecified?
Do not assign a missing or altered signal to a particular DOCK3 isoform: the supplied record lists 0 isoforms, and the A08904 caption does not identify its epitope (UniProt Q8IZD9 isoforms; datasheet A08904). DOCK3 contains an SH3 region at residues 6–67, a C2 DOCK-type region at 421–599, and a DOCKER region at 1228–1635, but those boundaries do not locate this antibody’s binding site (UniProt Q8IZD9 domains). For discordant sections, verify antibody identity and compare processing against a positive control processed alongside the sample (standard IHC practice). Seek epitope information before interpreting effects of variants or phosphoserine 1658 on recognition (UniProt Q8IZD9 modified residues; standard IHC practice).
How should I investigate DOCK3 by IF alongside the chromogenic IHC result?
Treat IF as a separate validation experiment: the A08904 example documents chromogenic staining in a paraffin section, with no IF procedure or IF-specific fixation result supplied (datasheet A08904). For a brain specimen, multiplex with a validated neuronal marker and assess whether DOCK3 signal follows the expected cytoplasmic and neuropil distribution (HPA tissue IHC; UniProt Q8IZD9 tissue specificity; standard IF practice). Choose spectrally separated fluorophores, including a longer-wavelength channel if tissue autofluorescence obscures the target, and inspect unstained controls (standard IF practice). Because DOCK3 is cytosolic and has no transmembrane segment, test permeabilisation sufficient for intracellular antibody access while checking morphology and background (HPA subcellular; UniProt Q8IZD9 topology; standard IF practice).
How can I distinguish diffuse DOCK3 staining from assay background?
Compare the section with a no-primary control and a processed positive section before calling diffuse color DOCK3 signal (standard IHC practice). The A08904 example used 10% goat serum block, 2 μg/ml primary antibody overnight at 4°C, and DAB detection; these are documented starting conditions for that example (datasheet A08904). Check whether background follows tissue folds, edges, or areas of poor morphology, and adjust blocking, washes, or primary concentration one variable at a time (standard IHC practice). Include a peroxidase block and review the no-primary section for endogenous enzyme activity when using HRP and DAB (standard chromogenic IHC practice).
How should I score DOCK3 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical region and cell population before scoring, since the reported DOCK3 signal includes CNS cytoplasm and neuropil (HPA tissue IHC; UniProt Q8IZD9 tissue specificity). For cellular staining, report the percentage of positive cells and an H-score using intensity categories 0–3, giving a possible score of 0–300 (standard IHC scoring practice). For neuropil, quantify chromogen-positive area or optical density within matched regions instead of forcing a cell count (standard image-analysis practice). Normalize cellular counts to all eligible cells and area measurements to the analyzed tissue area; hold staining batch, threshold, and excluded regions consistent across comparisons (standard IHC quantification practice).
What would make an apparent DOCK3-positive area unconvincing?
A convincing result should fit the expected cytoplasmic pattern and the sampled cell type; reported high signal occurs in cerebral-cortex neuronal cells, while colon endocrine cells are reported as not detected (HPA tissue IHC). Do not treat the A08904 colon-cancer image as evidence that every normal colon cell should stain, because its caption describes a cancer section (datasheet A08904). Nuclear-only color, edge-restricted staining, or signal concentrated in necrotic tissue warrants review with morphology and a no-primary control (HPA tissue IHC: cytoplasmic expression in CNS; standard IHC practice). If DAB persists without primary antibody, investigate endogenous peroxidase or other detection background before assigning a DOCK3-positive score (standard chromogenic IHC practice).
Boster reagents
Best DOCK3 / Dedicator of cytokinesis protein 3 IHC Antibodies
A08904 has chromogenic IHC images from paraffin-embedded human colon and lung cancer sections (catalog IHC captions). No IF/ICC images are supplied (catalog images).
Real IHC data
Anti-MOCA/DOCK3 Antibody ®
Cat # A08904
A08904 is listed for IHC and shows staining in paraffin-embedded human colon and lung cancer sections (catalog applications; catalog IHC captions). Its listed reactivity is Human, Mouse and Rat, but the supplied IHC images show human tissue only (catalog reactivity; catalog IHC captions).
Which to pick: Choose A08904 for tissue IHC: it is a rabbit antibody with human paraffin-section IHC images and a recommended concentration of 2–5 μg/ml (catalog host; catalog IHC captions; datasheet: 2–5 μg/ml). The captions do not report a fixative (catalog IHC captions). There is no IF/ICC recommendation because A08904 has no listed IF/ICC application or image; Mouse and Rat are listed as reactive species, but their tissue IHC performance is not shown here (catalog applications; catalog images; catalog reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8IZD9 (DOCK3_HUMAN, Dedicator of cytokinesis protein 3).
- Human Protein Atlas. DOCK3 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. DOCK3 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. DOCK3 antibody validation summary (2 antibodies).
- Dock3 protects myelin in the cuprizone model for demyelination. Cell death & disease 2014 — PMC4454328.
- Dock3 interaction with a glutamate-receptor NR2D subunit protects neurons from excitotoxicity. Molecular brain 2013 — PMC3652797.
- PubMed PMID:10854253 — UniProt-cited evidence.
- PubMed PMID:14569117 — UniProt-cited evidence.
- PubMed PMID:9205841 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DOCK3 (Dedicator of cytokinesis protein 3). Expected localisation Cytoplasmic staining in CNS (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A08904). Validated IHC antibody: A08904 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebral cortex (Neuronal cells); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet A08904); 2. Fixation: Image fixative and duration unreported (datasheet A08904); 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 A08904); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08904); 8. Primary antibody: Rabbit anti-DOCK3, 2-5 μg/ml (datasheet A08904); 9. Primary incubation: Overnight at 4 °C (datasheet A08904). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A08904); verify before use.; Caveat: Staining has medium concordance with RNA data (HPA tissue IHC); Isoform / epitope: No listed isoforms or processing; epitope site unspecified (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q8IZD9, HPA tissue IHC · Enhanced, HPA ICC-IF · cytosol., Datasheet A08904, PMC4454328. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DOCK3/immunohistochemistry-ihc-guide.