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DOK7 / Protein Dok-7 · IHC design guide
Design Immunohistochemistry for DOK7
Plan DOK7 IHC in paraffin sections using heart muscle as a strong positive tissue (HPA tissue IHC). This guide covers staining patterns, tissue controls, fixation consistency and chromogenic detection with the IHC-validated antibody (datasheet A05165-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DOK7 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 most tissues (HPA tissue IHC) | |
| Staining pattern | Strong cytoplasmic staining in cardiomyocytes (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05165-1) | |
| Positive control | Heart muscle+4 more · see all | |
| Negative control | Adipose tissue+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) | |
| Caveat | Presumed off-target binding; restaining pending (HPA tissue IHC) | |
| Regulation | Expression enriched in heart and skeletal muscle (UniProt) | |
| Isoform / epitope | Four isoforms; check epitope coverage across variants (UniProt) |
Section 1
Recommended DOK7 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is supplemented by one published DOK7 staining protocol using mouse tumor sections (PMC10756110).
| Sample | Paraffin-embedded human skeletal muscle tissue; fixative not specified (datasheet A05165-1) |
| Fixation | Image fixative and duration unreported (datasheet A05165-1); 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 A05165-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05165-1) |
| Primary antibody | Rabbit anti-DOK7, 2-5 μg/ml (datasheet A05165-1) |
| Primary incubation | Overnight at 4 °C (datasheet A05165-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A05165-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DOK7-positive staining in cardiomyocytes of heart muscle (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 for the catalog antibody (datasheet A05165-1); the published protocol used heated citrate with a different antibody (PMC10756110).
Section 2
What Is the Expected DOK7 Staining Pattern?
DOK7 should give predominantly cytoplasmic staining in tissue sections, with the clearest supplied positive example in heart cardiomyocytes (HPA: cytoplasmic expression in most tissues; High in cardiomyocytes). Enrichment near neuromuscular junctions is biologically plausible, but is not an established routine IHC pattern here (UniProt Q18PE1: accumulates at neuromuscular junctions). DOK7 has no transmembrane segment (UniProt Q18PE1: topology). HPA rates its tissue IHC profile Approved while noting presumed off target binding and pending restaining (HPA: tissue IHC reliability).
| Cytoplasmic chromogen is prominent in heart cardiomyocytes, with interpretable cellular boundaries and little staining in the negative control. | This matches the strongest listed tissue result: High cardiomyocyte staining (HPA: heart muscle IHC). Interpret the compartment as tissue cytoplasm; neuromuscular junction accumulation is a separate UniProt annotation, not proof that ordinary heart sections must show a junctional pattern (UniProt Q18PE1: subcellular location). |
| Staining is confined to nuclei, or forms a uniform surface rim, in otherwise readable heart sections. | Neither is the reported dominant tissue IHC pattern (HPA: cytoplasmic expression in most tissues). Review morphology and controls before calling it DOK7. A nuclear signal alone is not conclusive evidence of artefact because the separate ICC-IF dataset reports enhanced nucleoplasmic localization (HPA: subcellular ICC-IF). |
| Alveolar cells or adipocytes stain strongly while heart cardiomyocytes show little or no signal. | This reverses the supplied tissue comparison (HPA: alveolar cells and adipocytes Not detected; cardiomyocytes High). Consider cross-reactivity, nonspecific detection, or section handling; the unexpected stain cannot be assigned to DOK7 from morphology alone, especially while the tissue profile awaits restaining (HPA: reliability description). |
| A diffuse brown haze covers several structures without a clear cellular compartment. | The result cannot be scored against the reported cytoplasmic pattern (HPA: tissue IHC profile). In chromogenic IHC, nonspecific antibody binding or endogenous detection activity can create background (standard IHC practice). Compare the matched detection-only control and inspect whether chromogen follows cells rather than section-wide deposits. |
| Heart cardiomyocytes remain unstained although tissue architecture and counterstain are intact. | This is a failed positive control for the listed IHC pattern, not evidence that DOK7 is absent from all heart tissue (HPA: cardiomyocytes High; UniProt Q18PE1: heart protein expression). Check the catalog antibody's IHC-P instructions and the staining run before interpreting weaker tissues. |
💡Expected DOK7 appearanceCall a positive IHC result when heart cardiomyocytes show clear, predominantly cytoplasmic staining consistent with the listed High level; isolated nuclear, indiscriminate surface, or staining concentrated in listed negative cells needs control-based review (HPA: tissue IHC profile; heart cardiomyocytes High; alveolar cells and adipocytes Not detected).
| Tissue and cell choice | Heart cardiomyocytes provide the strongest listed positive comparison; adipocytes and lung alveolar cells are listed as Not detected (HPA: tissue IHC). Heart and skeletal muscle are preferential expression sites in the protein record, but the supplied HPA list gives no skeletal-muscle IHC staining level (UniProt Q18PE1: tissue specificity). |
| Antibody validation and reliability | HPA059449 is Approved for IHC, whereas its Enhanced designation applies to ICC; HPA062780 has no listed IHC status (HPA: antibody validation). The tissue profile is Approved but notes presumed off target binding and pending restaining, so unexpected cells need independent control-based assessment (HPA: reliability description). |
| Compartment and topology | Tissue IHC is described as cytoplasmic (HPA: tissue IHC profile). UniProt places DOK7 at the cell membrane and synapse, with neuromuscular junction accumulation and no transmembrane segment (UniProt Q18PE1: location and topology). These annotations do not justify demanding a continuous membrane rim in a paraffin section. |
| Isoforms and epitope coverage | UniProt lists 4 isoforms (UniProt Q18PE1: isoforms). An antibody could recognize a region shared by some isoforms or restricted to one; no epitope map is supplied here. Therefore, isoform-specific staining or absence cannot be inferred from this record, and any such claim requires antibody documentation. |
| Retrieval and processing limits | No target-specific antigen-retrieval condition or fixation sensitivity is provided (supplied UniProt and HPA records). UniProt lists no signal peptide, propeptide, glycosylation sites, or modified residues for this record (UniProt Q18PE1: processing and modifications); these facts do not predict paraffin-section retrieval performance. |
| Situation | Likely cause | Next action |
|---|---|---|
| Heart positive control has no specific stain. | The result conflicts with the listed High cardiomyocyte signal (HPA: heart muscle IHC); an ineffective staining run is possible (standard IHC practice). | Confirm tissue identity and cardiomyocyte preservation, then follow the catalog antibody's IHC-P instructions for retrieval, dilution, and detection; include a run control. No DOK7-specific retrieval setting is supplied. |
| Diffuse chromogen obscures cytoplasmic detail. | Background can arise from nonspecific binding or endogenous enzyme activity in chromogenic IHC (standard IHC practice). HPA's cytoplasmic profile cannot be evaluated through a section-wide haze (HPA: tissue IHC profile). | Compare detection-only and other appropriate negative controls; review blocking, reagent dilution, washes, and endogenous activity blocking as relevant to the detection system (standard IHC practice). |
| Strong staining appears in adipocytes or alveolar cells. | Both are listed as Not detected, so strong signal there is discordant and may reflect nonspecific staining (HPA: adipose tissue and lung IHC). The HPA tissue profile also notes presumed off target binding (HPA: reliability description). | Check cell identity and negative controls, then compare with a heart positive section in the same run. Report the discordance rather than assigning the unexpected cells as DOK7-positive. |
| Only nuclei or a sharp cell-surface rim stain in heart. | The dominant reported tissue pattern is cytoplasmic (HPA: tissue IHC profile). A nuclear ICC-IF location is reported separately and does not validate nuclear-only paraffin IHC (HPA: subcellular ICC-IF). | Recheck cellular morphology, counterstain, and control sections; assess whether the signal is reproducible with an independently validated approach before treating the compartment as target-specific. |
| Heart stains, but a second tissue gives weak or mixed signal. | The supplied tissue levels differ by cell type: heart cardiomyocytes are High, several glandular or neural cell types Medium, and liver hepatocytes Low (HPA: tissue IHC). A whole-section average can hide that distribution. | Score the named cell population and compartment within each tissue, using the same run controls. Describe weak or mixed staining at cell level rather than extrapolating the heart intensity to every tissue. |
| IF/ICC Q&A: why does a nuclear signal differ from the IHC expectation? | HPA reports enhanced nucleoplasmic and supported mitochondrial localization in ICC-IF, while tissue IHC is described as cytoplasmic (HPA: subcellular ICC-IF; tissue IHC profile). The datasets use different applications. | Interpret each result within its application and validation evidence. Do not use the ICC-IF location alone to reclassify a nuclear-only paraffin IHC result; consult the separate IF/ICC guide for that application. |
Experiment evidence
IHC & IF images showing DOK7 staining
A05165-1 · IHCIHC analysis of DOK7 using anti-DOK7 antibody (A05165-1). DOK7 was detected in a paraffin-embedded section of human skeletal muscle tissue.
A05165-1 · IHCIHC analysis of DOK7 using anti-DOK7 antibody (A05165-1). DOK7 was detected in a paraffin-embedded section of rat heart tissue. Heat mediate
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000175920-DOK7/tissue/cerebral+cortex#imid_16272795
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000175920-DOK7/tissue/colon#imid_16272737
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000175920-DOK7/tissue/liver#imid_16272722
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000175920-DOK7/tissue/kidney#imid_16272723
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA059449; green = DOK7, blue = nucleus, red = microtubules.
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA059449; green = DOK7, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DOK7 IHC & IF
🧪Run heart muscle first: cardiomyocytes should stain (HPA: High in cardiomyocytes). Use adipose tissue as the negative tissue (HPA: adipocytes Not detected); on the heart slide, compare cardiomyocytes with adjacent noncardiomyocyte cells as an internal background reference, without assuming those cells are DOK7-negative.
Positive control tissue: Heart muscle (Cardiomyocytes, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show DOK7 in MCF-7, Rh30, U2OS, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls (selected-SKU caption: rabbit primary), plus DOK7 knockout tissue or a validated peptide-block control if available (standard IHC specificity controls). Quench endogenous peroxidase and check pigment-related background in heart sections before interpreting DAB staining (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence; the selected-SKU paraffin-section caption also leaves the fixative unreported (selected-SKU caption). Heat retrieval with EDTA at pH 8.0 preceded skeletal-muscle staining at 2 μg/ml, but retrieval dependency has not been established (selected-SKU caption). Paraffin IHC is documented by that caption; whether frozen-section IHC or IF is easier is unreported, and heart pigment or endogenous peroxidase can complicate chromogenic scoring (standard IHC practice).
HPA tissue IHC evidence for DOK7
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Presumed off target binding observed and disregarded. Pending restaining.). 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 |
|---|---|---|---|---|
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Lung | Alveolar cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced DOK7 IHC Tips
Use the catalog antibody’s paraffin-section IHC conditions as the starting point, then interpret DOK7 staining against muscle controls and compartment-specific evidence.
Which antigen retrieval conditions should I start with for DOK7 chromogenic IHC?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections stained with A05165-1 (datasheet A05165-1). Keep heating and cooling conditions consistent across the test section and a skeletal muscle control, where this antibody produced staining under the stated retrieval condition (catalog caption A05165-1; standard IHC practice). If signal is weak, check section adhesion, antibody incubation, and detection reagents before changing retrieval (standard IHC practice). If retrieval remains suspect, compare a modestly longer heating interval with the original condition on adjacent sections, watching for tissue damage and increased background (standard IHC practice).
How should I troubleshoot weak DOK7 staining when fixation history is uncertain?
The selected paraffin-section caption does not state a fixative, so target-specific DOK7 sensitivity to fixation is unknown (catalog caption A05165-1). Record the fixative and fixation duration for each specimen, then compare sections processed with the same retrieval and detection conditions (standard IHC practice). Run a skeletal muscle control alongside the test section because that tissue was stained with this antibody at 2 μg/ml after EDTA retrieval at pH 8.0 (catalog caption A05165-1). If both sections are weak, investigate processing, retrieval, primary incubation, and detection before attributing the result to fixation (standard IHC practice).
Where should convincing DOK7 staining appear in muscle sections?
Assess staining in the context of muscle fibers and neuromuscular junctions, where DOK7 accumulates (UniProt Q18PE1 localisation). DOK7 is annotated at the cell membrane and synapse despite having no transmembrane segment, so a crisp transmembrane outline is not required for a plausible result (UniProt Q18PE1 localisation and topology). Cytoplasmic staining can also be plausible, although HPA reports cytoplasmic expression in most tissues and flags presumed off-target binding in its tissue series (HPA tissue IHC). Compare the test section with a skeletal muscle control and inspect whether signal follows intact structures rather than damaged edges (catalog caption A05165-1; standard IHC practice).
Could DOK7 isoforms or epitope accessibility explain uneven IHC staining?
DOK7 has 4 annotated isoforms, while the supplied antibody caption does not identify the recognized epitope (UniProt Q18PE1 isoforms; catalog caption A05165-1). The protein contains a PH domain at residues 4–109 and an IRS-type PTB domain at 105–210, but those coordinates alone cannot establish which isoforms this antibody detects (UniProt Q18PE1 domains; standard immunoassay interpretation). DOK7 has no annotated glycosylation sites or modified residues in the supplied record, so do not assign uneven staining to a specific modification (UniProt Q18PE1 processing). Compare adjacent sections under identical retrieval conditions and seek an independently mapped epitope before making isoform-specific claims (standard IHC practice).
How should I investigate DOK7 localisation with multiplex IF?
For the separate IF/ICC workflow, pair DOK7 with a skeletal muscle marker to identify the expected cell type, and consider MUSK to examine the reported interaction at neuromuscular junctions (UniProt Q18PE1 tissue specificity, subunit and localisation; standard IF practice). Choose spectrally separated fluorophores and place the weaker signal in a channel with low tissue autofluorescence, checking single-stain controls for bleed-through (standard IF practice). DOK7 lacks a transmembrane segment and interacts with the cytoplasmic part of MUSK, but the antibody epitope is unspecified (UniProt Q18PE1 topology and subunit; catalog caption A05165-1). Optimise permeabilisation for access to an intracellular epitope while preserving junctional structure, and verify staining against an appropriate negative control (standard IF practice).
What should I check when DOK7 DAB staining is diffuse or widespread?
First compare a no-primary control with the stained section to identify signal from detection reagents or endogenous peroxidase (standard IHC practice). Include a peroxidase-blocking step before DAB development and check whether the secondary reagent reacts with the specimen independently of the primary antibody (standard IHC practice). The catalog image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a 30-minute secondary incubation at 37°C (catalog caption A05165-1). If background persists, titrate primary concentration or incubation time while retaining a skeletal muscle positive control; interpret broad staining cautiously because HPA flags presumed off-target binding (standard IHC practice; HPA tissue IHC).
How should I quantify DOK7 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scoring compartment before analysis, then report an H-score or the percentage of positive muscle fibers using the same threshold across sections (standard IHC practice). If junctions are identifiable, report positive junction density per mm² of evaluable muscle area and document how junctions were recognized (standard IHC practice; UniProt Q18PE1 localisation). Normalise counts to viable tissue area or the number of evaluable fibers, and exclude folds, section edges, and damaged regions by a prespecified rule (standard IHC practice). Keep retrieval, DAB development, imaging, and threshold settings consistent, and include a skeletal muscle control across staining runs (catalog caption A05165-1; standard IHC practice).
How can I distinguish true DOK7 staining from section artefacts?
Give greatest weight to reproducible signal in intact muscle structures, especially near neuromuscular junctions, where DOK7 is reported to accumulate (UniProt Q18PE1 localisation; standard IHC practice). Treat staining confined to cut edges, folds, or necrotic areas as suspect and compare it with an adjacent section and a no-primary control (standard IHC practice). Diffuse DAB signal that survives primary omission suggests endogenous enzyme activity or detection background, while staining in an unexpected compartment needs independent support (standard IHC practice). HPA reports cytoplasmic tissue staining but also notes presumed off-target binding, and its IF record places DOK7 mainly in the nucleoplasm; these observations warrant cautious interpretation rather than automatic acceptance of either pattern (HPA tissue IHC; HPA subcellular).
Boster reagents
Best DOK7 / Protein Dok-7 IHC Antibodies
A05165-1 has real paraffin-section IHC images in human skeletal muscle and rat heart (catalog IHC captions). No IF image is supplied (catalog IF image alts).
Real IHC data
Anti-DOK7 Antibody ®
Cat # A05165-1
A05165-1 has IHC images of paraffin-embedded human skeletal muscle and rat heart (catalog IHC captions). It lists IHC and human, mouse and rat reactivity (catalog applications; reactivity); IF/ICC validation is not listed (catalog applications; IF image alts).
Which to pick: For tissue IHC, choose A05165-1, a rabbit polyclonal listed for IHC (catalog host; dilution_raw; applications); its paraffin-section images used EDTA pH 8 retrieval and 2 μg/ml primary antibody, and the fixative is unreported (catalog IHC captions). For IF/ICC, A05165-1 has no listed IF/ICC application or IF image (catalog applications; IF image alts), so it requires separate validation. For cross-species IHC, A05165-1 lists human, mouse and rat reactivity (catalog reactivity), with paraffin-section IHC images for human and rat (catalog IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q18PE1 (DOK7_HUMAN, Protein Dok-7).
- Human Protein Atlas. DOK7 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. DOK7 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the mitochondria..
- Human Protein Atlas. DOK7 antibody validation summary (2 antibodies).
- A prognostic model based on necroptosis-related genes for prognosis and therapy in bladder cancer. BMC urology 2023 — PMC9883845.
- DOK7, a target of miR-299-5p, suppresses the progression of bladder cancer. Aging 2023 — PMC10756110.
- ELF5 and DOK7 regulation in anti-estrogen treated cells and tumors. Cancer cell international 2016 — PMC4754800.
- Impaired signaling for neuromuscular synaptic maintenance is a feature of Motor Neuron Disease. Acta neuropathologica communications 2022 — PMC9040261.
- PubMed PMID:16794080 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DOK7 (Protein Dok-7). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05165-1). Validated IHC antibody: A05165-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Heart muscle (Cardiomyocytes); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human skeletal muscle tissue; fixative not specified (datasheet A05165-1); 2. Fixation: Image fixative and duration unreported (datasheet A05165-1); 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 A05165-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05165-1); 8. Primary antibody: Rabbit anti-DOK7, 2-5 μg/ml (datasheet A05165-1); 9. Primary incubation: Overnight at 4 °C (datasheet A05165-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Presumed off-target binding; restaining pending (HPA tissue IHC); Isoform / epitope: Four isoforms; check epitope coverage across variants (UniProt); Regulation: Expression enriched in heart and skeletal muscle (UniProt). Evidence: UniProt Q18PE1, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A05165-1, PMC10756110. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DOK7/immunohistochemistry-ihc-guide.