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MYO1D / Unconventional myosin-Id · IHC design guide
Design Immunohistochemistry for MYO1D
Plan chromogenic MYO1D IHC on paraffin sections with the catalog antibody’s 1:100–1:300 dilution range (datasheet: 1:100–1:300). Assess cytoplasmic staining and use controls to check specificity, given presumed off-target binding (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MYO1D 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 | Medium cytoplasmic staining in glandular and neuronal cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Appendix+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 paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target binding; verify staining specificity (HPA tissue IHC) | |
| Regulation | Brain has the highest reported expression (UniProt) | |
| Isoform / epitope | 0 reported isoforms; chain spans residues 2–1006 (UniProt) |
Section 1
Recommended MYO1D IHC & IF Protocols
The catalog antibody protocol is supplemented by published IHC methods for TNBC tissue microarrays (PMC10957690) and colorectal cancer sections (PMC8927999).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A09126-1) |
| Fixation | Image fixative and duration unreported (datasheet A09126-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-MYO1D, 1:100 - 1:300 (datasheet A09126-1) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MYO1D-positive staining in glandular cells of appendix (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen retrieval); the colorectal cancer method reports 7 min at high heat (PMC8927999).
Section 2
What Is the Expected MYO1D Staining Pattern?
MYO1D should appear predominantly in the cytoplasm, with possible enrichment near the cell cortex and puncta resembling transport vesicles (UniProt O94832: subcellular location; HPA tissue IHC: cytoplasmic expression in most tissues). HPA reports medium staining in several glandular and neuronal cell populations (HPA tissue IHC). Its tissue IHC rating is Approved, but staining has low consistency with RNA data and remains pending external verification (HPA tissue IHC: reliability). MYO1D has no transmembrane segment (UniProt O94832: topology).
| Cytoplasmic chromogen in colon or duodenal glandular cells, with nuclei comparatively clear (HPA tissue IHC: Medium in these cells). | This fits the reported tissue pattern and cytoplasmic location (HPA tissue IHC; UniProt O94832: subcellular location). Compare the signal with an appropriate control on the same run; a matching compartment alone cannot establish antibody specificity because HPA flags presumed off-target binding (HPA tissue IHC: reliability). |
| Cytoplasmic staining in cerebellar Purkinje cells or cerebral cortex or hippocampal neurons (HPA tissue IHC: Medium in these cells). | These are reported positive cell populations and can help assess whether the run detects the expected pattern (HPA tissue IHC). Record the stained cell type as well as intensity: UniProt describes high expression in brain, whereas the HPA entries report staining at the level of specified cells (UniProt O94832: tissue specificity; HPA tissue IHC). |
| Predominantly nuclear staining, or a continuous cell-surface outline without appreciable cytoplasmic signal. | Neither is the principal reported location (HPA subcellular ICC-IF: cytosol; UniProt O94832: subcellular location and no transmembrane segment). Check the compartment against controls and staining quality before calling it MYO1D; cortex-associated signal can lie close to a membrane without indicating a transmembrane protein (UniProt O94832: subcellular location and topology). |
| Strong staining in adipocytes, esophageal squamous cells, or lung alveolar cells (HPA tissue IHC: Not detected in these cells). | The cell-type mismatch warrants scrutiny for cross-reactivity or endogenous detection activity (HPA tissue IHC: reliability; general IHC practice). Do not treat lung as uniformly negative: UniProt describes high lung expression at the tissue level, while HPA specifically reports no staining in alveolar cells (UniProt O94832: tissue specificity; HPA tissue IHC). |
| Diffuse chromogen across cells and extracellular spaces, or no signal in colon or duodenal glandular cells (HPA tissue IHC: Medium in these cells). | Diffuse staining obscures compartment assessment; an absent expected signal makes the run inconclusive (general IHC practice). Review background controls, detection, antigen retrieval and the catalog antibody's IHC-P instructions before assigning biological meaning (general IHC practice). HPA's Approved rating does not resolve a failed or noisy local run (HPA tissue IHC: reliability). |
💡Expected MYO1D appearanceCall a result consistent when medium cytoplasmic staining is visible in reported glandular or neuronal cells, with possible cortex-associated or punctate emphasis; isolated nuclear staining or strong signal in HPA-undetected cell types is suspect (HPA tissue IHC; UniProt O94832: subcellular location).
| Which tissue and cell type are being scored? | HPA reports Medium staining in colon and duodenal glandular cells and in several neuronal populations, but Not detected in adipocytes and lung alveolar cells (HPA tissue IHC). Score the specified cells, not the entire section. UniProt's whole-tissue expression ranking cannot predict staining in every cell type (UniProt O94832: tissue specificity). |
| How certain is the antibody-based pattern? | The tissue IHC rating is Approved, with low staining–RNA consistency, external verification pending, and presumed off-target binding noted and disregarded in its assessment (HPA tissue IHC: reliability). The listed antibody HPA054744 has IHC Approved status; no Enhanced IHC status is supplied (HPA antibodies). Treat unexpected staining as provisional until controls support it. |
| Could a membrane-adjacent or punctate pattern be expected? | UniProt places MYO1D at the cell cortex near the apical membrane and in cytoplasmic puncta reminiscent of transport vesicles (UniProt O94832: subcellular location). Its lack of a transmembrane segment argues against interpreting a sharp, exclusive surface outline as its expected pattern (UniProt O94832: topology). |
| IF/ICC Q&A: where should fluorescence appear? | HPA's approved ICC-IF location is the cytosol (HPA subcellular ICC-IF). Assess cytosolic signal against the cell boundary and nucleus; UniProt also describes cortex-associated and vesicle-like punctate localization (UniProt O94832: subcellular location). This location guidance does not supply an IF/ICC protocol or validate an IHC-P result (HPA subcellular ICC-IF; general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| No chromogen appears in colon or duodenal glandular cells (HPA tissue IHC: Medium). | A run-level detection or retrieval problem is possible; the tissue result alone cannot identify the failed step (general IHC practice). | Confirm tissue preservation and positive-control staining, then check the catalog antibody's IHC-P retrieval, dilution and detection instructions (general IHC practice). Do not infer MYO1D absence from this run. |
| The whole section develops a diffuse haze that masks cytoplasmic detail. | Nonspecific reagent binding, excess primary antibody or incomplete washing can raise background (general IHC practice). | Compare the no-primary control, review blocking and washes, and adjust primary antibody dilution using the validated IHC-P protocol (general IHC practice). Score only after cell boundaries and intracellular staining are distinguishable. |
| A brown precipitate appears in negative-control tissue or in cells reported Not detected (HPA tissue IHC). | Endogenous enzyme activity, if enzyme-based detection is used, or nonspecific binding can mimic chromogen signal (general IHC practice). | Inspect no-primary and detection-only controls and apply the detection system's endogenous-activity blocking procedure where relevant (general IHC practice). Reassess whether signal tracks the reported cytoplasmic cell pattern (HPA tissue IHC). |
| Staining is predominantly nuclear or forms only a crisp cell-surface rim. | The distribution conflicts with the reported cytosolic and cortex-associated locations; artefact or off-target binding is possible (HPA subcellular ICC-IF; UniProt O94832: subcellular location; HPA tissue IHC: reliability). | Check morphology, counterstain and controls, then compare a reported positive cell population on the same run (general IHC practice; HPA tissue IHC). Avoid scoring the atypical compartment as confirmed MYO1D. |
| Lung alveolar cells stain strongly despite an otherwise clean run (HPA tissue IHC: Not detected). | HPA's cell-specific result and UniProt's high whole-lung expression describe different levels of observation; neither establishes alveolar-cell positivity (HPA tissue IHC; UniProt O94832: tissue specificity). | Document the exact stained cell type and compare control sections before interpreting the signal (general IHC practice). HPA's low staining–RNA consistency and off-target caveat make this unexpected result provisional (HPA tissue IHC: reliability). |
| The run shows variable intensity among areas of a reported positive tissue (HPA tissue IHC). | Uneven section quality or reagent coverage can create an apparent gradient (general IHC practice); HPA's Medium category does not guarantee uniform staining in every cell (HPA tissue IHC). | Inspect morphology and within-run control staining, repeat the affected section if technical unevenness is evident, and record intensity by cell population rather than averaging the whole tissue (general IHC practice). |
Experiment evidence
IHC & IF images showing MYO1D staining
A09126-1 · IHCImmunohistochemistry analysis of paraffin-embedded human brain tissue, using MYO1D Antibody. The picture on the right is blocked with the sy
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000176658-MYO1D/tissue/cerebral+cortex#imid_15740043
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000176658-MYO1D/tissue/colon#imid_15739985
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000176658-MYO1D/tissue/liver#imid_15739978
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000176658-MYO1D/tissue/kidney#imid_15739987
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA054744; green = MYO1D, blue = nucleus, red = microtubules.
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA054744; green = MYO1D, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MYO1D IHC & IF
🧪Run cerebellum first: Purkinje cells should stain at a medium level (HPA: cerebellum, Purkinje cells, Medium). Use adipose tissue as a negative tissue, where adipocytes are not detected (HPA: adipose tissue, adipocytes, Not detected); cells on the cerebellar slide without specific cytoplasmic staining should remain at background, but their identity as MYO1D-negative cells is unverified (UniProt O94832: cytoplasmic localization).
Positive control tissue: Appendix (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MYO1D in MCF-7, PC-3, U2OS, NIH 3T3, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control and a concentration-matched isotype control appropriate to the primary antibody’s host species and clonality (standard IHC practice). Use a MYO1D knockout sample if available, or a peptide-block control where the immunizing peptide is available (standard IHC practice; A09126-1 caption: peptide-blocked brain image); quench endogenous peroxidase for chromogenic detection, and assess neuronal autofluorescence if using IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative in the selected A09126-1 paraffin-section brain caption is unreported (A09126-1 caption). Antigen-retrieval dependence is unreported, so optimize retrieval empirically for paraffin IHC; the supplied evidence does not establish whether frozen sections or IF are easier (A09126-1 caption; HPA: ICC-IF cytosol localization). In cerebellum, neuronal autofluorescence can complicate IF interpretation, so compare against the no-primary control (standard IF practice).
HPA tissue IHC evidence for MYO1D
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. Pending external verification. Presumed off target binding observed and disregarded.). 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MYO1D IHC Tips
Troubleshoot MYO1D staining in paraffin sections by checking retrieval, compartment, controls and cell-specific signal before comparing samples.
How should I retrieve MYO1D in paraffin sections with weak chromogenic staining?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval setting). Let sections cool in buffer, then apply the catalog antibody and compare a retrieved section with an otherwise identical section that skipped heating (standard IHC practice). If staining remains weak, test a different retrieval buffer only as a fallback, holding section thickness, antibody concentration and detection conditions constant (standard IHC practice). Evaluate cytoplasmic staining in neuronal cells of cerebral cortex or hippocampus, where medium staining is reported (HPA tissue IHC), and reject a retrieval change that mainly increases diffuse background.
Could fixation explain variable MYO1D staining across paraffin blocks?
Target-specific fixation sensitivity for MYO1D is unknown because the selected brain tissue caption identifies paraffin embedding but does not report a fixative (A09126-1 caption). Record each block’s fixative and fixation duration, and compare blocks processed under matched conditions before changing the antibody dilution (standard IHC practice). If prolonged fixation is suspected, run the stated citrate pH 6.0, 95–98 °C, 20 min retrieval consistently across a small comparison set (page retrieval setting). Judge recovery by cell-specific cytoplasmic staining and preserved morphology; the peptide-blocked brain image supports antibody competition in that preparation but cannot establish a fixation limit (A09126-1 caption).
What MYO1D staining pattern should count as plausible in brain IHC?
Score predominantly cytoplasmic signal, with possible cortical or punctate emphasis, because MYO1D is annotated in the cytoplasm, cell cortex, dendrites and early endosomes (UniProt O94832 localisation). Medium staining in cerebral cortical and hippocampal neuronal cells provides tissue-level context for selecting fields (HPA tissue IHC). Compare neuronal somata and processes with adjacent cells on the same section, using a counterstain to verify boundaries (standard IHC practice). A strong nucleus-only pattern should trigger checks of retrieval, detection background and antibody specificity before interpretation, since the approved cell-image location is cytosol (HPA subcellular).
How do I assess whether epitope accessibility affects MYO1D IHC?
Document the immunogen region if the catalog record provides it; the supplied caption gives no epitope coordinates for A09126-1 (A09126-1 caption). MYO1D has a motor domain at residues 9–695, two IQ motifs at 699–719 and 721–741, and a TH1 region at 812–1005 (UniProt O94832 domains). Its record lists 0 isoforms and modified residues including phosphoserine 200 and phosphotyrosine 536 (UniProt O94832). Without a mapped epitope, do not attribute staining differences to a particular domain or modification; compare retrieval conditions and peptide competition while keeping detection constant (standard IHC practice; A09126-1 caption).
How can IF help check an ambiguous chromogenic MYO1D pattern?
Use IF on a matched specimen to examine whether MYO1D signal overlaps a marker identifying the cell type being scored in IHC; choose the marker independently of MYO1D staining (standard IF practice). Include single-label controls and select fluorophores away from the tissue’s strongest autofluorescence, checking each channel separately before interpreting overlap (standard IF practice). MYO1D lacks a transmembrane segment and is annotated in cytoplasm, cortex and endosomes, so permeabilise sufficiently for intracellular epitope access while preserving cell boundaries (UniProt O94832 topology and localisation; standard IF practice). Compare the resulting cytosolic pattern with the approved cytosol annotation, then return to the chromogenic section for scoring (HPA subcellular).
What should I check when MYO1D chromogenic staining is diffuse?
Inspect a no-primary control alongside the test section to identify signal from the detection system, and check whether endogenous peroxidase was blocked before chromogen development (standard IHC practice). Titrate the primary antibody and shorten chromogen development one variable at a time while keeping retrieval at citrate pH 6.0, 95–98 °C for 20 min (page retrieval setting; standard IHC practice). Compare diffuse staining with the expected cytoplasmic distribution rather than accepting uniformly dark tissue as positive (HPA tissue IHC). The selected brain image includes peptide blocking, which can support a competition check for that preparation but does not resolve every source of background (A09126-1 caption).
How should I quantify MYO1D staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and tissue region before reviewing treatment groups, then use identical retrieval, illumination and chromogen development across slides (standard IHC practice). Record the percentage of positive cells and staining intensity, or calculate an H-score from intensity categories 0–3 multiplied by their percentages (standard IHC scoring practice). Normalise cell counts to the number of eligible cells, or report positive-cell density per mm² of viable scored tissue when cell counts are impractical (standard IHC practice). Keep neuronal and glandular populations separate because their reported staining refers to different cell types, and exclude folds, edges and necrotic areas consistently (HPA tissue IHC; standard IHC practice).
When is a MYO1D-positive IHC result convincing rather than artefactual?
A convincing result has cell-resolved cytoplasmic staining in a plausible population, such as neuronal cells in cerebral cortex, and remains interpretable against adjacent tissue (HPA tissue IHC; UniProt O94832 localisation). Review nucleus-only staining, tissue-edge gradients and necrotic areas as possible artefacts before calling a section positive (HPA subcellular; standard IHC practice). Check the no-primary control for endogenous enzyme or detection signal, and use the peptide-blocked brain image as limited competition evidence for the selected antibody preparation (standard IHC practice; A09126-1 caption). Interpret tissue comparisons cautiously because the HPA tissue profile is approved yet reports low agreement with RNA and presumed off-target staining (HPA tissue IHC reliability).
Boster reagents
Best MYO1D / Unconventional myosin-Id IHC Antibodies
A09126-1 has a real IHC image from paraffin-embedded human brain tissue and lists IF/ICC applications and human, mouse, and rat reactivity (catalog: A09126-1 image caption, applications, reactivity).
Real IHC data
Anti-MYO1D/Myosin Id Antibody
Cat # A09126-1
A09126-1 will render with an IHC image of paraffin-embedded human brain tissue and a peptide-blocked comparison (catalog: A09126-1 IHC image caption). It lists IHC, IF, ICC, and ELISA applications and human, mouse, and rat reactivity; no IF image is supplied (catalog: A09126-1 applications, reactivity, IF image alts).
Which to pick: For tissue IHC, choose A09126-1: its own image caption shows paraffin-embedded human brain tissue, but does not report the fixative (catalog: A09126-1 IHC image caption). For IF/ICC, A09126-1 is the listed option, although no IF image is supplied (catalog: A09126-1 applications, IF image alts). For cross-species work, A09126-1 is a rabbit polyclonal antibody listing human, mouse, and rat reactivity (catalog: A09126-1 host, dilution_raw, reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O94832 (MYO1D_HUMAN, Unconventional myosin-Id).
- Human Protein Atlas. MYO1D tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MYO1D subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. MYO1D antibody validation summary (1 antibodies).
- Identification and validation of neutrophils-related subtypes and prognosis model in triple negative breast cancer. Journal of cancer research and clinical oncology 2024 — PMC10957690.
- The Role of miR-217-5p in the Puromycin Aminonucleoside-Induced Morphological Change of Podocytes. Non-coding RNA 2022 — PMC9229466.
- Identification of metastasis-associated exoDEPs in colorectal cancer using label-free proteomics. Translational oncology 2022 — PMC8927999.
- PubMed PMID:9872452 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:18669648 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MYO1D (Unconventional myosin-Id). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A09126-1 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet A09126-1); 2. Fixation: Image fixative and duration unreported (datasheet A09126-1); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-MYO1D, 1:100 - 1:300 (datasheet A09126-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Presumed off-target binding; verify staining specificity (HPA tissue IHC); Isoform / epitope: 0 reported isoforms; chain spans residues 2–1006 (UniProt); Regulation: Brain has the highest reported expression (UniProt). Evidence: UniProt O94832, HPA tissue IHC · Approved, HPA ICC-IF · cytosol., Datasheet A09126-1, PMC10957690, PMC8927999. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MYO1D/immunohistochemistry-ihc-guide.