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MYZAP / Myocardial zonula adherens protein · IHC design guide
Design Immunohistochemistry for MYZAP
Start chromogenic MYZAP IHC-P with the catalog antibody at 5 μg/mL (datasheet: IHC-P). Assess cardiomyocyte intercalated discs and lung macrophages, and interpret staining cautiously because presumed off-target binding was observed (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MYZAP 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 | Predominantly intercalated discs; low Z-disc signal (UniProt) | |
| Staining pattern | Cardiomyocyte intercalated discs; lung macrophage staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| 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 paraffin-section fixation consistent (standard IHC practice; not target-specific); assess effects empirically. | |
| Caveat | Presumed off-target binding may complicate scoring (HPA tissue IHC) | |
| Regulation | Heart muscle-enriched RNA (HPA tissue RNA) | |
| Isoform / epitope | 11 isoforms; mature chain 21–466; check epitope coverage (UniProt) |
Section 1
Recommended MYZAP IHC & IF Protocols
The catalog antibody protocol uses citrate pH 6.0 retrieval (page retrieval rule). One published MYZAP IHC protocol reports low-pH retrieval for FFPE sections (PMC8656111).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A13349); 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-MYZAP, 5 μg/mL (datasheet A13349) |
| 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 | MYZAP-positive staining in cardiomyocytes of heart muscle (HPA tissue IHC: High). HPA tissue profile: Most abundant expression in intercalated discs and lung macrophages. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule); use the study’s low-pH retrieval when reproducing its protocol (PMC8656111).
Section 2
What Is the Expected MYZAP Staining Pattern?
MYZAP is predominantly at cardiomyocyte intercalated discs, with lower levels at sarcomeric Z discs; it is also associated with the cytoskeleton and cell junctions and has no transmembrane segment (UniProt P0CAP1). Expect strong staining in heart cardiomyocytes and lung macrophages (HPA: High in both). Interpret tissue staining cautiously: HPA rates its IHC evidence Approved, with medium RNA–staining consistency and presumed off-target binding (HPA: tissue IHC reliability).
| Strong, sharply defined staining at borders between cardiomyocytes. | This matches the predominant intercalated-disc localisation reported for MYZAP (UniProt P0CAP1: subcellular location) and the high cardiomyocyte staining observed in heart muscle (HPA: High in cardiomyocytes). Assess the pattern within intact heart architecture; intensity alone cannot establish that every stained structure contains MYZAP (HPA: tissue IHC reliability). |
| Prominent nuclear staining dominates cardiomyocytes. | A nuclear-dominant pattern conflicts with the reported junctional and cytoskeletal localisation (UniProt P0CAP1: subcellular location). Treat it as suspect and compare the antibody control and tissue morphology before scoring it as MYZAP; chromogen deposits or nonspecific binding can mislead localisation (general IHC practice). |
| Strong staining appears in cells outside the expected tissue pattern. | Check the cell identity before calling cross-reactivity: lung macrophages also stain strongly, while placental trophoblasts, cerebellar Purkinje cells and parathyroid glandular cells stain at medium levels (HPA: tissue IHC). An unexpected cell population may reflect nonspecific or endogenous detection activity; HPA also cautions that its tissue assay targets protein from more than one gene (HPA: tissue IHC reliability). |
| Broad, even chromogen covers cells and surrounding tissue. | Diffuse staining without a discernible cardiomyocyte junctional pattern is difficult to interpret against the predominant intercalated-disc localisation (UniProt P0CAP1: subcellular location). Review negative detection controls, blocking and washing for general background; do not assign widespread staining to MYZAP from colour alone (general IHC practice). |
| Heart cardiomyocytes show no discernible staining. | This conflicts with high cardiomyocyte staining in the HPA heart section (HPA: High in cardiomyocytes). First check tissue preservation, control performance and the IHC run; absent signal in one section does not establish absent MYZAP expression (general IHC practice). |
💡Expected MYZAP appearanceCall a positive heart result when high cardiomyocyte staining highlights intercalated discs, with at most weaker Z-disc staining (HPA: High in cardiomyocytes; UniProt P0CAP1: subcellular location); a dominant nuclear or indiscriminately diffuse pattern is suspect (UniProt P0CAP1: subcellular location; general IHC practice).
| Tissue and cell choice | Heart cardiomyocytes provide a high-staining reference, but lung macrophages are also high; adipocytes in adipose tissue and adrenal glandular cells are reported as not detected (HPA: tissue IHC). Score the specified cell population, since a whole-section positive or negative label can conceal which cells stained (general IHC practice). |
| Subcellular pattern | MYZAP is detected predominantly at cardiomyocyte intercalated discs and at low levels on sarcomeric Z discs (UniProt P0CAP1: subcellular location). Use compartment and tissue architecture alongside intensity when judging a result; membrane-associated staining should not be assumed to indicate a transmembrane protein (UniProt P0CAP1: topology). |
| Evidence and antibody specificity | HPA lists HPA039827 IHC as Approved, reports medium consistency with RNA expression, and warns of presumed off-target binding and detection of protein from more than one gene (HPA: antibody validation; HPA: tissue IHC reliability). A plausible staining pattern therefore supports interpretation but does not by itself prove molecular specificity. |
| Isoforms and antibody epitope | UniProt lists 11 MYZAP isoforms and a processed chain spanning residues 21–466 (UniProt P0CAP1: isoforms; processing). No epitope position is supplied here, so isoform coverage and any effect of processing on this antibody cannot be determined. Avoid attributing a missed stain to either mechanism without antibody-specific evidence. |
| Antigen retrieval and fixation | These are general IHC variables that can affect staining and should be assessed against run controls (general IHC practice). Neither source supplied here establishes a MYZAP-specific retrieval requirement or fixation sensitivity; do not infer one from the UniProt topology or HPA tissue staining levels. |
| IF/ICC expected pattern? | UniProt places MYZAP at cardiomyocyte intercalated discs and, more weakly, Z discs (UniProt P0CAP1: subcellular location). HPA supplies no ICC-IF images or main subcellular location for this record, and lists no ICC status for HPA039827 (HPA: subcellular; antibody validation). This evidence supports a localisation expectation, not an ICC-IF validation or protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Heart positive control is blank. | The run may have failed, or the selected section may not show assessable cardiomyocytes (general IHC practice). High cardiomyocyte staining is expected in the HPA heart reference (HPA: tissue IHC). | Confirm cardiomyocyte-rich morphology and review the run's positive control, primary-antibody step, detection reagents and counterstain before interpreting a sample as negative (general IHC practice). |
| Signal is present but intercalated discs are indistinct. | Background or tissue detail may obscure the junctional pattern (general IHC practice); the expected predominant compartment is the intercalated disc (UniProt P0CAP1: subcellular location). | Compare intact neighbouring cardiomyocytes and the negative detection control; assess washing, blocking and chromogen development as general IHC variables before scoring localisation (general IHC practice). |
| Lung macrophages stain, but heart is negative. | HPA reports high staining in both populations, so the lung result alone does not resolve whether the heart result reflects tissue, run or antibody behaviour (HPA: tissue IHC; tissue IHC reliability). | Repeat assessment with an adequate heart section and run controls. Record each cell type separately, and avoid using macrophage staining alone to claim MYZAP-specific detection (HPA: tissue IHC reliability; general IHC practice). |
| Many unexpected cells stain strongly. | Cell misidentification, nonspecific binding or endogenous detection activity may contribute (general IHC practice); HPA flags presumed off-target binding and detection of protein from more than one gene (HPA: tissue IHC reliability). | Identify the stained cells by morphology, compare HPA's cell-specific tissue pattern and inspect negative detection controls before assigning the signal to MYZAP (HPA: tissue IHC; general IHC practice). |
| A proposed negative-control tissue shows staining. | HPA's ‘not detected’ calls apply to named cells, such as adipocytes in adipose tissue and adrenal glandular cells, rather than every cell in those sections (HPA: tissue IHC). | Score the named cell population and its compartment; then compare a negative detection control and the heart positive reference to judge whether the staining is interpretable (HPA: tissue IHC; general IHC practice). |
| Nuclei or the whole section appear uniformly brown. | That distribution does not match the reported predominant cardiomyocyte junctional pattern (UniProt P0CAP1: subcellular location) and may reflect background, endogenous detection activity or excessive chromogen development (general IHC practice). | Check the negative detection control, counterstain balance and chromogen development; score MYZAP only where cellular identity and a credible compartmental pattern can be resolved (general IHC practice). |
Experiment evidence
IHC & IF images showing MYZAP staining
A13349 · IHCImmunohistochemistry of MYOZAP in human lung tissue with MYOZAP antibody at 5 μg/mL.
A13349 · IFImmunofluorescence of MYOZAP in human lung tissue with MYOZAP antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000263155-MYZAP/tissue/cerebral+cortex#imid_10327977
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000263155-MYZAP/tissue/colon#imid_10327895
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000263155-MYZAP/tissue/liver#imid_10327888
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000263155-MYZAP/tissue/kidney#imid_10327905
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MYZAP IHC & IF
🧪Run heart muscle first: cardiomyocytes should stain (HPA: High in cardiomyocytes). Use adipose tissue as the negative tissue, with adipocytes expected to be unstained (HPA: Not detected in adipocytes); on the heart slide, assess morphologically identified noncardiomyocytes for background staining rather than assuming they are MYZAP-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 carries no ICC-IF cell line for MYZAP; derive a cell-line control from the positive tissue's cell type (Cardiomyocytes) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only), an isotype control matched to the primary antibody’s host species and clonality, and MYZAP-knockout tissue or a validated peptide-block control (standard IHC practice). Check endogenous peroxidase activity from residual blood in heart sections during chromogenic detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence; the selected A13349 lung IHC caption also does not state its fixative (selected-SKU caption). The need for antigen retrieval in paraffin sections is unreported, so assess retrieval conditions empirically (standard IHC practice). The supplied evidence does not establish whether frozen sections or IF are easier for MYZAP; residual blood in heart sections can produce peroxidase-related background (standard IHC practice).
HPA tissue IHC evidence for MYZAP
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene. 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 |
|---|---|---|---|---|
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
| Lung | Macrophages | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MYZAP IHC Tips
Troubleshoot chromogenic MYZAP IHC by checking retrieval, controls, staining location and cell identity before comparing staining scores.
How should I retrieve MYZAP in paraffin sections when staining is weak?
Use citrate buffer at pH 6.0 for heat-induced antigen retrieval at 95–98 °C for 20 min before chromogenic IHC (page retrieval rule). Include heart muscle on each run and check cardiomyocyte intercalated discs for the expected pattern (HPA tissue IHC; UniProt P0CAP1 localisation). If staining remains weak, compare a longer retrieval time on adjacent sections while keeping antibody concentration and detection conditions constant (standard IHC practice). Check tissue integrity and background alongside signal: excessive heating can damage sections or expose nonspecific binding, so choose conditions that preserve both morphology and junctional detail (standard IHC practice).
Could fixation explain absent MYZAP staining in my paraffin sections?
Target-specific fixation sensitivity is unknown from the supplied evidence; the catalog lung image reports 5 µg/mL antibody but does not state its fixative (selected A13349 tissue-IHC caption). Record fixation method and duration for each specimen, then compare similarly processed sections before changing retrieval or antibody concentration (standard IHC practice). Include heart muscle as a positive control and assess intercalated-disc staining alongside section morphology (HPA tissue IHC; UniProt P0CAP1 localisation). If control and test sections both fail, check the staining run with a positive control before attributing the result to fixation; do not infer fixation sensitivity from tissue expression or protein topology (standard IHC practice).
Where should convincing MYZAP chromogenic staining appear?
In heart muscle, look first for staining at cardiomyocyte intercalated discs, with weaker staining possible at sarcomeric Z discs (UniProt P0CAP1 localisation; HPA tissue IHC). MYZAP is also annotated in the cytoplasm, cytoskeleton, cell membrane and cell junction, so evaluate the pattern in its tissue context rather than calling all cytoplasmic colour positive (UniProt P0CAP1 localisation). The catalog image documents staining in human lung at 5 µg/mL, while HPA reports abundant staining in lung macrophages (selected A13349 tissue-IHC caption; HPA tissue IHC). Compare stained cells with the counterstained morphology and a negative reagent control before assigning compartment or cell identity (standard IHC practice).
How do MYZAP isoforms affect the interpretation of a negative IHC result?
MYZAP has 11 listed isoforms, but the supplied evidence does not map the catalog antibody's epitope to any of them (UniProt P0CAP1 isoforms; selected A13349 tissue-IHC caption). The record lists residues 1–20 as a signal sequence and 21–466 as the chain; epitope coverage cannot be inferred from those boundaries alone (UniProt P0CAP1 processing). Check the antibody's stated immunogen or epitope, if available, before treating a negative section as absence of every isoform (standard IHC interpretation). Compare heart-muscle control staining and tissue morphology across matched runs, and report the reagent and scoring method used so another reader can judge what the negative result supports (HPA tissue IHC; standard IHC practice).
How can I check MYZAP localisation by IF alongside chromogenic IHC?
For a separate IF experiment, pair MYZAP with a cardiomyocyte marker such as cardiac troponin T and assess whether junctional signal lies in the expected cells (UniProt P0CAP1 localisation; standard IF practice). Choose fluorophores after inspecting unstained tissue for autofluorescence, and favour channels with clear separation from that background (standard IF practice). MYZAP has no annotated transmembrane segment, but the antibody's epitope side is unspecified; compare mild permeabilisation with an unpermeabilised condition if epitope access is uncertain (UniProt P0CAP1 topology; standard IF practice). Include single-label and no-primary controls when judging colocalisation, because the lung IHC caption alone does not establish IF performance (selected A13349 tissue-IHC caption; standard IF practice).
How do I reduce diffuse brown staining without losing MYZAP signal?
Start with a no-primary control, a peroxidase-block check and inspection for pigment or precipitated chromogen before changing the MYZAP antibody (standard chromogenic IHC practice). Optimise blocking, washes and antibody concentration on matched sections, using the catalog image's 5 µg/mL only as a documented lung-IHC reference point (selected A13349 tissue-IHC caption; standard IHC practice). Keep DAB development time consistent across comparisons, since stronger development can increase apparent background as well as signal (standard chromogenic IHC practice). Review heart-muscle intercalated discs and lung macrophages separately: both are reported staining sites, so cell-specific signal should not automatically be called diffuse background (HPA tissue IHC; UniProt P0CAP1 localisation).
What should I score when comparing MYZAP IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Predefine whether the outcome is cardiomyocyte junctional staining, positive-cell fraction or stained-cell density, and use the same definition for every section (UniProt P0CAP1 localisation; standard IHC quantification practice). An H-score can combine intensity with the percentage of eligible cells, while macrophage-rich lung samples may be clearer as positive macrophages per mm² of viable tissue (HPA tissue IHC; standard IHC quantification practice). Normalise junctional measurements to assessed cardiomyocytes or evaluable junction length, and cell counts to viable tissue area or the relevant cell population (standard IHC quantification practice). Exclude folds, edges and necrotic regions by a rule set before reviewing group labels, and record the antibody concentration, retrieval and DAB timing (standard IHC quantification practice).
When is a brown MYZAP signal more likely artefact than true staining?
A convincing heart-muscle result follows cardiomyocyte intercalated discs, whereas isolated colour in an unexpected compartment needs corroboration before being assigned to MYZAP (UniProt P0CAP1 localisation; HPA tissue IHC). Lung macrophage staining can be relevant, but identify the cells morphologically and compare no-primary controls before interpreting brown material as antigen signal (HPA tissue IHC; standard IHC practice). Discount staining concentrated at section edges, folds or necrotic regions, and check endogenous peroxidase and DAB controls when colour appears outside intact cells (standard chromogenic IHC practice). HPA rates its tissue staining Approved while noting medium RNA agreement and presumed off-target binding, so confirm consequential findings with an independent reagent or method (HPA tissue IHC; standard IHC validation practice).
Boster reagents
Best MYZAP / Myocardial zonula adherens protein IHC Antibodies
A13349 is listed for IHC-P and IF, with images of human lung tissue for both applications; catalog reactivity includes human, mouse, and rat (catalog: applications and reactivity; IHC and IF image captions).
Real IHC data
Anti-MYOZAP MYZAP Antibody
Cat # A13349
A13349 has an IHC image of human lung tissue at 5 μg/mL (IHC image caption). A13349 also has an IF image of human lung tissue at 20 μg/mL; its listed reactivity is human, mouse, and rat (IF image caption; catalog: reactivity).
Which to pick: For tissue IHC, choose A13349: IHC-P is listed, and its own IHC image shows human lung tissue at 5 μg/mL; the fixative is unreported (catalog: applications; IHC image caption). For IF, choose A13349 at a starting concentration of 20 μg/mL; separate ICC validation and clone type are unreported (datasheet: IF starting concentration; catalog: applications and clone). For mouse or rat samples, A13349 lists reactivity with both species, while the supplied IHC and IF images show human tissue (catalog: reactivity; IHC and IF image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P0CAP1 (MYZAP_HUMAN, Myocardial zonula adherens protein).
- Human Protein Atlas. MYZAP tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MYZAP subcellular location (ICC-IF): Highest expression in HEL: 83.8 nTPM.
- Human Protein Atlas. MYZAP antibody validation summary (1 antibodies).
- A biallelic loss-of-function variant in MYZAP is associated with a recessive form of severe dilated cardiomyopathy. Cold Spring Harbor molecular case studies 2022 — PMC9528970.
- GRINL1A Complex Transcription Unit Containing GCOM1, MYZAP, and POLR2M Genes Associates with Fully Penetrant Recessive Dilated Cardiomyopathy. Frontiers in genetics 2021 — PMC8656111.
- Subclinical oral involvement in patients with endemic pemphigus foliaceus. Dermatology practical & conceptual 2018 — PMC6246068.
- PubMed PMID:15233991 — UniProt-cited evidence.
- PubMed PMID:20093627 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MYZAP (Myocardial zonula adherens protein). Expected localisation Predominantly intercalated discs; low Z-disc signal (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A13349 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Heart muscle (Cardiomyocytes); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A13349); 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-MYZAP, 5 μg/mL (datasheet A13349); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); assess effects empirically.; Caveat: Presumed off-target binding may complicate scoring (HPA tissue IHC); Isoform / epitope: 11 isoforms; mature chain 21–466; check epitope coverage (UniProt); Regulation: Heart muscle-enriched RNA (HPA tissue RNA). Evidence: UniProt P0CAP1, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in HEL: 83.8 nTPM, Datasheet A13349, PMC8656111. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MYZAP/immunohistochemistry-ihc-guide.