This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
MYBPC3 / Myosin-binding protein C, cardiac-type · IHC design guide
Design Immunohistochemistry for MYBPC3
Plan MYBPC3 staining in paraffin sections using heart cardiomyocyte cytoplasmic staining as the expected pattern (HPA tissue IHC). This guide covers tissue controls and interpretation alongside the catalog antibody’s IHC dilution of 0.5–1 μg/ml (datasheet A01078-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MYBPC3 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 | Cardiomyocyte cytoplasm (HPA tissue IHC) | |
| Staining pattern | Heart cardiomyocytes show cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A01078-1) | |
| Positive control | Heart muscle+1 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 | Testis spermatogonia also stain strongly (HPA tissue IHC) | |
| Regulation | Expression regulation not specified (UniProt) | |
| Isoform / epitope | 2 isoforms; check epitope coverage (UniProt) |
Section 1
Recommended MYBPC3 IHC & IF Protocols
The catalog antibody's IHC-P protocol (datasheet: A01078-1) is followed by a published MYBPC3 IHC protocol using paraffin-embedded left ventricle (PMC8924128).
| Sample | Paraffin-embedded rat heart tissue; fixative not specified (datasheet A01078-1) |
| Fixation | Image fixative and duration unreported (datasheet A01078-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A01078-1) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01078-1) |
| Primary antibody | Rabbit anti-MYBPC3, 0.5-1μg/ml (datasheet A01078-1) |
| Primary incubation | Overnight at 4 °C (datasheet A01078-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01078-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MYBPC3-positive staining in cardiomyocytes of heart muscle (HPA tissue IHC: High). HPA tissue profile: Highly selective cytoplasmic expression in heart myocytes. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet: A01078-1); the published protocol does not report retrieval conditions (PMC8924128).
Section 2
What Is the Expected MYBPC3 Staining Pattern?
MYBPC3 is a thick filament associated protein of striated muscle A bands and has no transmembrane segment (UniProt Q14896: function, topology). In paraffin sections, expect strong cytoplasmic staining in heart cardiomyocytes (HPA: High in cardiomyocytes). HPA describes the tissue profile as highly selective for heart myocytes and rates its IHC evidence Enhanced because antibody staining agrees closely with RNA expression (HPA: tissue IHC reliability).
| Strong cytoplasmic staining follows the cardiomyocyte fibers in heart muscle. | This fits the reported high cardiomyocyte signal (HPA: High in cardiomyocytes) and MYBPC3’s association with striated muscle A bands (UniProt Q14896: function). Fine sarcomeric detail may depend on section orientation and image resolution (general IHC practice); its absence alone does not overturn a convincing cytoplasmic pattern. |
| Signal is concentrated in nuclei, along cell surfaces, or in extracellular material. | Those compartments do not fit the expected cardiomyocyte cytoplasmic profile (HPA: tissue IHC profile) or a protein with no transmembrane segment (UniProt Q14896: topology). Treat the pattern as possible artefact and compare it with the negative control before assigning MYBPC3 positivity (general IHC practice). |
| Prominent staining appears in an unexpected cell population or a reported negative tissue. | Consider cross-reactivity or endogenous detection activity (general IHC practice). HPA reports no detection in adipocytes of adipose tissue and glandular cells of adrenal gland, but reports high staining in testis spermatogonia (HPA: tissue IHC). Therefore, testis signal alone is not evidence of a false positive. |
| Brown precipitate covers many structures without a clear cardiomyocyte pattern. | Diffuse background prevents a reliable cell specific call (general IHC practice). Review the negative control and whether background follows tissue edges, damaged areas, or multiple compartments (general IHC practice). A widespread haze should not be scored as the selective heart myocyte staining reported by HPA (HPA: tissue IHC profile). |
| Heart cardiomyocytes show no detectable signal. | A negative result in the expected positive tissue conflicts with HPA’s high cardiomyocyte staining (HPA: High in cardiomyocytes). Check that cardiomyocytes are present and assess the control, antibody dilution, retrieval, and detection steps before interpreting the sample as MYBPC3 negative (general IHC practice). |
💡Expected MYBPC3 appearanceCall a section positive when cardiomyocyte cytoplasm shows strong, fiber associated staining (HPA: High in cardiomyocytes; UniProt Q14896: function); isolated nuclear, surface, or widespread nonspecific staining is suspect (general IHC practice).
| Target location and topology | MYBPC3 is associated with thick filaments in striated muscle A bands and has no transmembrane segment (UniProt Q14896: function, topology). Interpret the expected signal within cardiomyocyte cytoplasm, rather than as a membrane outline (HPA: tissue IHC profile). |
| Tissue context | Heart muscle is RNA tissue enriched, and cardiomyocytes stain at a high level (HPA: tissue IHC). High staining is also listed for testis spermatogonia (HPA: tissue IHC); use the named cell population when interpreting that tissue. |
| Isoforms and modified residues | UniProt lists two isoforms and multiple modified residues, including phosphorylated serines (UniProt Q14896: isoforms, modified residues). The supplied sources do not map the IHC antibody epitopes, so they do not establish whether either feature changes staining. |
| Evidence for the IHC pattern | HPA rates the tissue IHC profile Enhanced and lists two rabbit antibodies with Enhanced IHC status (HPA: tissue IHC reliability; HPA: HPA040147, HPA043898). This supports the reported tissue pattern, without establishing a specific retrieval condition or fixation sensitivity. |
| IF/ICC evidence limit | HPA provides no main subcellular location or cell line ICC/IF images for this target (HPA: subcellular record). The cytoplasmic IHC expectation should not be presented as a validated IF/ICC pattern; consult the separate IF/ICC guide for that application. |
| Situation | Likely cause | Next action |
|---|---|---|
| Heart cardiomyocytes are blank while the negative control is clean. | The expected positive cell population has not been demonstrated (HPA: High in cardiomyocytes); a processing, retrieval, antibody, or detection step may have failed (general IHC practice). | Confirm cardiomyocytes are present, then check reagent activity, antibody dilution, retrieval, and detection against the catalog antibody’s IHC-P instructions (general IHC practice). Do not infer MYBPC3 absence from this slide alone. |
| Signal is mostly nuclear or outlines cardiomyocyte membranes. | This differs from the reported cytoplasmic profile (HPA: tissue IHC profile) and from the absence of a transmembrane segment (UniProt Q14896: topology); background or mislocalised detection is possible (general IHC practice). | Compare with a negative control and a heart positive control, then review blocking, washes, and detection background (general IHC practice). Score only a convincing cardiomyocyte cytoplasmic pattern. |
| A reported negative tissue stains strongly. | Cross-reactivity or endogenous detection activity may contribute (general IHC practice). HPA reports no detection in adipose tissue adipocytes and adrenal gland glandular cells (HPA: tissue IHC). | Check the stained cell identity against HPA’s cell level entries, inspect a negative control, and review detection blocking (general IHC practice). Do not use testis spermatogonia as a presumed negative control: HPA reports high staining there (HPA: tissue IHC). |
| Brown haze obscures cell boundaries across the section. | Diffuse reagent background or endogenous detection activity can reduce contrast (general IHC practice); such a pattern does not match selective cardiomyocyte cytoplasmic staining (HPA: tissue IHC profile). | Inspect negative controls, tissue condition, blocking, wash stringency, and detection development (general IHC practice). Reassess only after the background permits a clear cell level call. |
| Heart signal is weak or uneven despite visible cardiomyocytes. | Section quality, retrieval, antibody concentration, or detection may contribute (general IHC practice). The supplied sources give no MYBPC3 specific fixation effect or optimal retrieval setting. | Compare matched heart sections and controls, then follow the catalog antibody’s IHC-P retrieval and dilution instructions (general IHC practice). Avoid attributing the result to MYBPC3 epitope masking without supporting evidence. |
| An IF/ICC image is being used to validate the paraffin IHC result. | HPA lists no ICC/IF cell line images or main subcellular location for MYBPC3 (HPA: subcellular record); its Enhanced rating here concerns IHC tissue evidence (HPA: tissue IHC reliability). | Judge the paraffin section against the heart cardiomyocyte IHC pattern and its controls (HPA: tissue IHC; general IHC practice). Evaluate IF/ICC separately using application specific evidence. |
Experiment evidence
IHC & IF images showing MYBPC3 staining
A01078-1 · IHCIHC analysis of MYBPC3 using anti-MYBPC3 antibody (A01078-1). MYBPC3 was detected in paraffin-embedded section of rat heart tissue. Heat med
A01078-1 · IHCIHC analysis of MYBPC3 using anti-MYBPC3 antibody (A01078-1). MYBPC3 was detected in paraffin-embedded section of human lung cancer tissue.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000134571-MYBPC3/tissue/cerebral+cortex#imid_10497171
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000134571-MYBPC3/tissue/colon#imid_10469739
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000134571-MYBPC3/tissue/liver#imid_10469740
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000134571-MYBPC3/tissue/kidney#imid_10469733
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MYBPC3 IHC & IF
🧪Run heart muscle first; cardiomyocytes should stain (HPA: High in cardiomyocytes). Use adipose tissue as the negative, focusing on adipocytes (HPA: Not detected in adipocytes); on the heart slide, neighboring non-cardiomyocytes should lack the cardiomyocyte staining pattern and serve as an internal background reference (HPA: High in cardiomyocytes).
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 MYBPC3; 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 and concentration-matched rabbit IgG isotype controls, matching primary clonality if known (caption: rabbit anti-MYBPC3). Use MYBPC3 knockout tissue as a biological negative, and quench endogenous peroxidase and check for endogenous biotin in heart tissue when using the reported biotin/SABC-DAB detection (caption: biotinylated secondary and SABC/DAB).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A01078-1 tissue-IHC caption does not state a fixative (caption: fixative unreported). The rat heart paraffin-section example used heat retrieval in citrate buffer at pH 6 for 20 minutes; this documents a working retrieval condition but does not establish that retrieval is required (caption: rat heart IHC). IHC on paraffin sections has a supplied example, while the supplied evidence provides no basis to conclude that frozen sections or IF are easier (caption: rat heart IHC; HPA: no ICC-IF images).
HPA tissue IHC evidence for MYBPC3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data.). 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 → |
| Testis | Spermatogonia cells | High | 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 → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MYBPC3 IHC Tips
Troubleshoot MYBPC3 chromogenic IHC in paraffin sections using the catalog antibody’s tissue image, tissue expression data, and the protein’s known structure and function.
How should I adjust retrieval when MYBPC3 staining is weak or uneven?
Start with heat mediated citrate retrieval at pH 6 for 20 minutes (datasheet A01078-1). The catalog antibody produced staining in a paraffin section of rat heart after this treatment, followed by 1 μg/ml primary antibody overnight at 4°C (datasheet A01078-1). If signal is weak, check heating consistency and section adhesion before cautiously varying retrieval duration on matched sections (standard IHC practice). Compare staining within cardiomyocyte cytoplasm and tissue morphology across conditions; excessive retrieval can damage sections and make localisation harder to judge (HPA tissue IHC; standard IHC practice). The caption does not state the fixative, so its retrieval result does not establish performance across fixation methods (datasheet A01078-1).
Could fixation explain a weak MYBPC3 result in paraffin sections?
Target specific fixation sensitivity is unknown because the selected paraffin section caption does not state its fixative (datasheet A01078-1). Record the fixative and processing history for each specimen, then compare matched heart sections processed consistently before attributing weak staining to fixation (standard IHC practice; HPA tissue IHC). Use the documented citrate retrieval at pH 6 for 20 minutes as the starting condition for this antibody (datasheet A01078-1). If fixation histories differ, assess morphology, staining distribution, and a positive control in each batch; differences alone cannot identify which processing step caused signal loss (standard IHC practice). MYBPC3’s phosphorylation sites do not establish fixation sensitivity (UniProt Q14896).
Where should credible MYBPC3 chromogenic staining appear within heart tissue?
Assess staining primarily in cardiomyocyte cytoplasm, where tissue IHC reports highly selective expression in heart myocytes (HPA tissue IHC). MYBPC3 is a thick filament associated protein in the crossbridge region of striated muscle A bands, so an organised myofibrillar pattern is biologically plausible when section quality permits its resolution (UniProt Q14896). Strong isolated nuclear or membrane outlined staining should prompt review of morphology and controls rather than immediate assignment to MYBPC3 (UniProt Q14896; standard IHC practice). Its record lists no transmembrane segment and no annotated subcellular location, while the available subcellular dataset provides no ICC/IF image confirmation (UniProt Q14896; HPA subcellular). Judge localisation within intact cells, away from folds and damaged edges (standard IHC practice).
Can this antibody distinguish MYBPC3 isoforms or phosphorylation states in tissue?
Do not interpret this chromogenic signal as isoform specific without an epitope map and isoform validation for the catalog antibody (standard IHC practice). The record lists 2 isoforms and several phosphorylation sites, including residues 275, 284, and 304, but the supplied tissue caption gives no epitope or phospho specificity (UniProt Q14896; datasheet A01078-1). MYBPC3 also contains repeated Ig like and fibronectin type III domains, making epitope location relevant when comparing reagents (UniProt Q14896). If a biological question requires one splice form or phosphorylation state, obtain direct antibody validation and appropriate controls before scoring sections; routine chromogenic intensity cannot resolve that distinction (standard IHC practice).
How would I check MYBPC3 localisation by multiplex immunofluorescence?
For a separate IF experiment, pair MYBPC3 with an independently validated cardiomyocyte marker and inspect signal within the same cells (HPA tissue IHC; standard IF practice). Select spectrally separated fluorophores after checking tissue autofluorescence and include single stain controls to assess bleed through (standard IF practice). Because MYBPC3 has no transmembrane segment and is associated with intracellular thick filaments, use permeabilisation appropriate for an intracellular epitope, then verify that myofibrillar morphology remains interpretable (UniProt Q14896; standard IF practice). The supplied subcellular dataset has no ICC/IF images, and the selected antibody caption documents chromogenic paraffin tissue IHC only; neither establishes IF performance or an IF fixation condition (HPA subcellular; datasheet A01078-1).
What should I troubleshoot when brown staining extends beyond cardiomyocytes?
First compare the suspect area with intact cardiomyocytes and a negative control, because the tissue profile describes highly selective cytoplasmic expression in heart myocytes (HPA tissue IHC; standard IHC practice). The documented workflow used 10% goat serum blocking, biotinylated goat anti rabbit secondary antibody, an avidin biotin detection complex, and DAB (datasheet A01078-1). Check secondary only controls, endogenous peroxidase blocking, washing, and DAB development when widespread brown signal appears; these are general chromogenic IHC troubleshooting steps, not MYBPC3 specific findings (standard IHC practice). Compare section edges, folds, and damaged tissue separately, since deposits there can obscure a cellular pattern (standard IHC practice).
How should I score MYBPC3 staining across heart sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define cardiomyocytes as the scoring population and keep tissue region, section quality, staining batch, and image settings consistent across samples (HPA tissue IHC; standard IHC practice). An H score combines the percentages of cells at ordinal intensity grades, whereas percentage positive cells measures prevalence and positive area per tissue area measures extent (standard IHC practice). Normalise the selected readout to the number of intact cardiomyocytes or evaluable myocardial area, and report excluded folds, edges, and damaged regions (standard IHC practice). Set thresholds using appropriate controls before comparing groups; a change in brown area can reflect altered cell content or section quality as well as altered MYBPC3 staining (standard IHC practice).
How can I distinguish true MYBPC3 signal from a staining artefact?
A credible result aligns with cytoplasmic staining in intact cardiomyocytes and, where resolved, the expected thick filament organisation (HPA tissue IHC; UniProt Q14896). Treat dominant nuclear or membrane outlined signal, staining restricted to section edges, and signal in necrotic regions as reasons to review controls and morphology (UniProt Q14896; standard IHC practice). A no primary control helps expose detection system staining, while adequate endogenous peroxidase blocking helps assess enzyme related brown deposits in a DAB assay (standard IHC practice). High staining is also reported in testis spermatogonia, so cell identity and tissue context matter when interpreting an unexpected positive outside heart (HPA tissue IHC).
Boster reagents
Best MYBPC3 / Myosin-binding protein C, cardiac-type IHC Antibodies
A01078-1 has real IHC images of paraffin sections from rat heart and human lung, mammary, and rectal cancer tissues (A01078-1 IHC image captions). No IF image is provided (catalog IF image alts).
Real IHC data
Anti-MYBPC3 Antibody ®
Cat # A01078-1
A01078-1 is listed for IHC and has images from rat heart and human lung, mammary, and rectal cancer paraffin sections (catalog applications; A01078-1 IHC image captions). Its listed reactivity is human, mouse, and rat, while the supplied IHC images show human and rat tissue (catalog reactivity; A01078-1 IHC image captions).
Which to pick: For tissue IHC, choose A01078-1 for paraffin sections matching its image evidence; the captions do not report the fixative (A01078-1 IHC image captions). For IF/ICC, no listed SKU has an IF/ICC application or IF image (catalog applications; catalog IF image alts). For work across species, A01078-1 is a rabbit polyclonal with listed human, mouse, and rat reactivity; its supplied IHC images show human and rat tissue (catalog host and dilution_raw; catalog reactivity; A01078-1 IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q14896 (MYPC3_HUMAN, Myosin-binding protein C, cardiac-type).
- Human Protein Atlas. MYBPC3 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. MYBPC3 subcellular location (ICC-IF): Highest expression in THP-1: 0.4 nTPM.
- Human Protein Atlas. MYBPC3 antibody validation summary (2 antibodies).
- Case Report: Identification of the First Synonymous Variant of Myosin Binding Protein C3 (c.24A>C, p.P8P) Altering RNA Splicing in a Cardiomyopathy and Sudden Cardiac Death Case. Frontiers in cardiovascular medicine 2022 — PMC8924128.
- MDM2 Regulation of HIF Signaling Causes Microvascular Dysfunction in Hypertrophic Cardiomyopathy. Circulation 2023 — PMC10691664.
- Targeting Cardiomyocyte PCNA and POLD1 Prevents Pathologic Myocardial Hypertrophy. Circulation research 2025 — PMC12509454.
- Expression patterns of cardiac myofilament proteins: genomic and protein analysis of surgical myectomy tissue from patients with obstructive hypertrophic cardiomyopathy. Circulation. Heart failure 2009 — PMC2765062.
- PubMed PMID:7744002 — UniProt-cited evidence.
- PubMed PMID:9048664 — UniProt-cited evidence.
- PubMed PMID:9562578 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MYBPC3 (Myosin-binding protein C, cardiac-type). Expected localisation Cardiomyocyte cytoplasm (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A01078-1). Validated IHC antibody: A01078-1 at 0.5-1μg/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 rat heart tissue; fixative not specified (datasheet A01078-1); 2. Fixation: Image fixative and duration unreported (datasheet A01078-1); 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: Citrate pH 6 , 20 min (datasheet A01078-1); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01078-1); 8. Primary antibody: Rabbit anti-MYBPC3, 0.5-1μg/ml (datasheet A01078-1); 9. Primary incubation: Overnight at 4 °C (datasheet A01078-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Testis spermatogonia also stain strongly (HPA tissue IHC); Isoform / epitope: 2 isoforms; check epitope coverage (UniProt); Regulation: Expression regulation not specified (UniProt). Evidence: UniProt Q14896, HPA tissue IHC · Enhanced, HPA cell-line RNA · Highest expression in THP-1: 0.4 nTPM, Datasheet A01078-1, PMC8924128. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MYBPC3/immunohistochemistry-ihc-guide.