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CSRP3 / Cysteine and glycine-rich protein 3 · IHC design guide
Design Immunohistochemistry for CSRP3
Plan chromogenic IHC for CSRP3 in paraffin sections using heart and skeletal muscle as positive tissues (HPA tissue IHC). The guide covers cytoplasmic staining expectations, fixation consistency, and interpretation of the two isoforms (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CSRP3 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 | Muscle cytoplasm observed; Z line expected (HPA tissue IHC; UniProt) | |
| Staining pattern | High cytoplasmic staining in cardiomyocytes and skeletal myocytes (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| 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 | Isoform coverage is unknown without antibody epitope data (UniProt; datasheet) | |
| Regulation | Isoform 2 is higher in muscle disease (UniProt) | |
| Isoform / epitope | Two isoforms; antibody epitope coverage is unspecified (UniProt; datasheet) |
Section 1
Recommended CSRP3 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet) with the published chromogenic IHC protocol for rat glioma tissue (PMC13223896).
| Sample | Paraffin-embedded mouse heart tissue; fixative not specified (datasheet A06522) |
| Fixation | Image fixative and duration unreported (datasheet A06522); 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 Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear 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-CSRP3, 1:50-1:200 (datasheet A06522) |
| 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 | CSRP3-positive staining in cardiomyocytes of heart muscle (HPA tissue IHC: High). HPA tissue profile: Selective cytoplasmic expression in heart and skeletal muscle. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min (page retrieval rule).
Section 2
What Is the Expected CSRP3 Staining Pattern?
CSRP3 should stain mainly the cytoplasm of cardiomyocytes in heart muscle and myocytes in skeletal muscle; both are reported as high (HPA: tissue IHC, Enhanced reliability). Sarcomeric Z-line association can add an organized striated appearance, while nuclear staining may occur (UniProt P50461: localization). CSRP3 has no transmembrane segment, so a membrane outline is not the expected dominant pattern (UniProt P50461: topology).
| Strong cytoplasmic staining in cardiomyocytes or skeletal myocytes, with an organized pattern where structures are resolved. | This matches the high staining reported in both cell types (HPA: tissue IHC). A Z-line-associated pattern is biologically plausible, but routine chromogenic sections may not resolve individual Z lines (UniProt P50461: localization; general IHC practice). |
| A membrane outline or extracellular-looking signal dominates; alternatively, nuclei stain strongly with little cytoplasmic signal. | Dominant membrane staining conflicts with the absence of a transmembrane segment and mainly cytoplasmic localization (UniProt P50461: topology and localization). Nuclear signal alone is not conclusive evidence of artifact because CSRP3 can shuttle into the nucleus (UniProt P50461: localization); check controls and the full pattern. |
| Strong staining appears in cells reported as negative, such as adipocytes in adipose tissue or glandular cells in adrenal gland. | These cell types were not detected in the HPA tissue survey (HPA: tissue IHC). Unexpected signal can reflect antibody cross-reactivity or endogenous detection activity (general IHC practice); compare the negative tissue, no-primary control and cellular distribution before assigning a cause. |
| Color spreads across tissue or the slide without a clear myocyte-specific cytoplasmic pattern. | Diffuse background does not match the selective cytoplasmic profile (HPA: tissue IHC). Uneven reagent coverage, inadequate blocking or insufficient washing are general IHC possibilities (general IHC practice); background alone cannot establish CSRP3 expression. |
| No convincing staining appears in heart cardiomyocytes or skeletal myocytes. | Those are the supplied high-staining reference cells (HPA: tissue IHC). A negative section therefore needs a technical check before a biological interpretation; inspect tissue integrity, assay controls, retrieval and antibody application (general IHC practice). |
💡Expected CSRP3 appearanceCall a positive result when cytoplasmic signal is strong in cardiomyocytes or skeletal myocytes (HPA: tissue IHC); isolated membrane outlines or widespread staining in HPA-negative cells should trigger a false-positive check (UniProt P50461: topology; HPA: tissue IHC; general IHC practice).
| Tissue and cell selection | Heart cardiomyocytes and skeletal myocytes are high-staining reference cells; the listed adipose adipocytes and adrenal glandular cells were not detected (HPA: tissue IHC). These comparisons help judge selectivity within the stated HPA survey. |
| Subcellular distribution | CSRP3 is mainly cytoplasmic and can associate with sarcomeric Z lines; it also shuttles to the nucleus (UniProt P50461: localization). Score the dominant pattern in its tissue context rather than treating every nuclear signal as impossible. |
| Antibody evidence | HPA042581 has Enhanced IHC validation (HPA: antibody validation). This supports its reported tissue pattern, but does not establish that every unexpected signal in a new section is specific (general IHC practice). |
| Isoforms and epitope | UniProt lists 2 CSRP3 isoforms; isoform 2 is expressed in striated muscle (UniProt P50461: isoforms and tissue specificity). The supplied evidence does not map the antibody epitope, so the IHC pattern cannot identify which isoform contributed signal. |
| IF/ICC Q&A: where should fluorescence appear? | Mainly in cytosol; nucleoplasm is an additional uncertain location in HPA ICC-IF images (HPA: subcellular ICC-IF). That uncertainty should not be converted into a requirement for nuclear staining in paraffin IHC. |
| Situation | Likely cause | Next action |
|---|---|---|
| Heart or skeletal muscle is blank despite an intact-looking section. | A technical failure remains possible because these myocytes are reported high in tissue IHC (HPA: tissue IHC); the image alone does not identify which step failed. | Check positive-control performance, antibody application, retrieval conditions and detection reagents using the established IHC workflow (general IHC practice). Reassess the myocytes before calling the sample CSRP3-negative. |
| Staining is largely nuclear, with weak or absent cytoplasmic signal. | Some nuclear localization is plausible, but CSRP3 is mainly cytoplasmic (UniProt P50461: localization). Counterstain overlap or nonspecific nuclear signal may complicate interpretation (general IHC practice). | Compare nuclear and cytoplasmic signal in the same cells and inspect a no-primary control (general IHC practice). Record limited nuclear signal separately; investigate a dominant nuclear-only pattern before accepting it. |
| Membranes or extracellular spaces look more positive than myocyte cytoplasm. | That distribution conflicts with CSRP3's mainly cytoplasmic localization and lack of a transmembrane segment (UniProt P50461: localization and topology). | Confirm the apparent compartment at suitable magnification and compare positive tissue with a no-primary control (general IHC practice). Treat a persistent membrane-dominant pattern as unexpected rather than scoring it as the expected CSRP3 result. |
| Cells in an HPA-negative tissue show substantial color. | The listed negative cell types were not detected in the HPA survey (HPA: tissue IHC). Cross-reactivity or endogenous detection activity are possible explanations, not diagnoses from appearance alone (general IHC practice). | Compare the staining with a no-primary control and the high-staining reference tissue (HPA: tissue IHC; general IHC practice). If signal persists only with primary antibody, assess whether its cell and compartment pattern is credible. |
| The whole section has diffuse color that obscures cell boundaries. | Diffuse background prevents assessment of the selective cytoplasmic pattern (HPA: tissue IHC). Blocking, washing and reagent handling can affect background in chromogenic IHC (general IHC practice). | Inspect the no-primary control, then review blocking, wash steps and chromogen development under the established assay conditions (general IHC practice). Score CSRP3 only after myocyte cytoplasm is distinguishable from background. |
| A faint positive-looking pattern appears, but the expected striations are unresolved. | Z-line association is reported for CSRP3, while routine section resolution and orientation can limit visible striation (UniProt P50461: localization; general IHC practice). Absence of visible lines alone does not negate cytoplasmic staining. | Judge signal against cardiomyocyte or skeletal-myocyte cytoplasm and matched background (HPA: tissue IHC; general IHC practice). Describe striations only when they are actually visible. |
Experiment evidence
IHC & IF images showing CSRP3 staining
A06522 · IHCImmunohistochemistry of paraffin-embedded mouse heart using CSRP3 Antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000129170-CSRP3/tissue/cerebral+cortex#imid_13161319
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000129170-CSRP3/tissue/colon#imid_13161261
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000129170-CSRP3/tissue/liver#imid_13161262
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000129170-CSRP3/tissue/kidney#imid_13161247
HPA · ICC-IF THP-1Human Protein Atlas immunofluorescence, THP-1, antibody HPA042581; green = CSRP3, blue = nucleus, red = microtubules.
HPA · ICC-IF THP-1Human Protein Atlas immunofluorescence, THP-1, antibody HPA042581; green = CSRP3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CSRP3 IHC & IF
🧪Run heart muscle first: cardiomyocytes should stain (HPA: High in cardiomyocytes; selected A06522 tissue-IHC caption: mouse heart). Use adipose tissue as the negative: adipocytes are not detected there (HPA: Not detected in adipocytes), while non-cardiomyocyte areas on the heart section should be assessed for background rather than assumed to be CSRP3-negative (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 ICC-IF images show CSRP3 in THP-1, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; an isotype control matched to the primary antibody’s host species, immunoglobulin class and antibody format; and CSRP3-knockout tissue as a biological negative (standard IHC practice). Quench endogenous peroxidase in heart sections before chromogenic detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A06522 paraffin-section caption does not state the fixative (selected A06522 tissue-IHC caption). That caption uses microwave retrieval in 10 mM PBS at pH 7.2 and an antibody dilution of 1:100; whether retrieval is required under other conditions is unreported (selected A06522 tissue-IHC caption). Whether frozen sections or IF are easier is unreported; heart muscle autofluorescence can complicate IF assessment (standard IF practice).
HPA tissue IHC evidence for CSRP3
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 → |
| Skeletal muscle | Myocytes | 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 CSRP3 IHC Tips
Troubleshoot CSRP3 staining in paraffin sections by checking retrieval, compartment, muscle cell identity, and controls before comparing signal intensity.
What retrieval conditions should I start with for CSRP3 IHC?
Use Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval setting). Let sections cool in buffer, then compare staining with an otherwise identical no-primary control to identify detection background (standard IHC practice). If staining is weak, test microwave retrieval in 10 mM PBS at pH 7.2, the condition reported for paraffin-embedded mouse heart with catalog antibody A06522 (A06522 tissue-IHC caption). Keep section thickness, detection chemistry, and development time constant while comparing conditions, and assess signal in cardiomyocytes rather than judging retrieval by overall tissue darkness (HPA tissue IHC; standard IHC practice).
How should I troubleshoot fixation-related loss of CSRP3 staining?
CSRP3-specific sensitivity to fixation is unknown from the supplied evidence; the paraffin-embedded mouse-heart caption does not state its fixative (A06522 tissue-IHC caption). Record the actual fixative and fixation duration for each specimen, and compare matched sections processed with the same retrieval and detection steps (standard IHC practice). If a batch stains weakly, check whether section age, retrieval, or detection also changed before assigning the loss to fixation (standard IHC practice). Include a previously staining heart section in the same run, and interpret a selective loss in test sections against that control without inferring a CSRP3-specific fixation mechanism (HPA tissue IHC; standard IHC practice).
Should CSRP3 staining appear in the cytoplasm or nucleus?
Expect predominantly cytoplasmic staining in cardiomyocytes and skeletal myocytes; HPA reports selective cytoplasmic expression in those tissues (HPA tissue IHC). CSRP3 also associates with the sarcomeric Z line and can shuttle between cytoplasm and nucleus, so a structured cytoplasmic pattern or limited nuclear signal is biologically plausible (UniProt P50461 subcellular). Assess nuclear staining separately from cytoplasmic staining, using a counterstain to define each compartment and a no-primary section to assess background (standard IHC practice). Widespread nuclear staining across unrelated cell types deserves scrutiny because HPA reports mainly cytosolic localisation and regards its additional nucleoplasmic localisation as uncertain (HPA subcellular).
Can this antibody distinguish CSRP3 isoforms or phosphorylation states in tissue?
Do not score an IHC pattern as isoform-specific without an antibody epitope mapped to sequence unique to one isoform; the supplied antibody caption gives no epitope information (A06522 tissue-IHC caption; standard IHC practice). CSRP3 has 2 isoforms, and isoform 2 is reported in striated muscle and at higher levels in specified neuromuscular diseases (UniProt P50461 isoforms; UniProt P50461 tissue specificity). Its LIM domains span residues 10–61 and 120–171, while reported phosphoserines occur at 95 and 153 (UniProt P50461 domains; UniProt P50461 modified residues). Neither total staining nor a change after retrieval identifies an isoform or phosphorylation state without an independently validated selective reagent (standard IHC practice).
How can I follow up a chromogenic CSRP3 pattern by immunofluorescence?
Treat IF as a separate assay: the supplied product image establishes paraffin-section chromogenic IHC conditions, not IF performance (A06522 tissue-IHC caption). Multiplex CSRP3 with a validated marker identifying cardiomyocytes or skeletal myocytes, and include single-stain controls to check channel bleed-through (HPA tissue IHC; standard IF practice). Choose a fluorophore and emission channel after checking tissue autofluorescence, and compare an unstained section under the same exposure settings (standard IF practice). Because CSRP3 lacks a transmembrane segment and is mainly cytoplasmic, use permeabilisation appropriate for an intracellular epitope, then optimise it against preservation of myofibrillar structure (UniProt P50461 topology; UniProt P50461 subcellular; standard IF practice).
What should I check when CSRP3 IHC stains many unrelated cells?
Start with a no-primary section and a peroxidase block when using horseradish-peroxidase detection; these distinguish detection background from primary-antibody staining (standard IHC practice). Compare the same run with heart or skeletal muscle, where myocytes show high staining, and with a listed negative tissue such as adipose tissue, where adipocytes were not detected (HPA tissue IHC). If both controls stain diffusely, review blocking, antibody concentration, washes, and chromogen development time before interpreting the test section (standard IHC practice). Use tissue morphology to flag deposits at folds, damaged edges, and necrotic areas, and avoid counting those deposits as cellular CSRP3 (standard IHC practice).
How should I quantify CSRP3 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define cardiomyocytes or skeletal myocytes as the scoring population before analysis, because HPA reports high staining in those cells (HPA tissue IHC). For chromogenic IHC, record the percentage of positive target cells and intensity categories 0–3, then calculate an H-score from 0–300 using identical thresholds across slides (standard IHC practice). If measuring stained area or optical density, normalise to viable target-cell area rather than whole-section area, which may vary in muscle content (standard IHC practice). Keep retrieval, counterstain, illumination, and chromogen development consistent, and report cytoplasmic and nuclear scores separately when both compartments are evaluated (UniProt P50461 subcellular; standard IHC practice).
How can I tell genuine CSRP3 staining from an artefact?
A convincing positive lies in muscle cells with predominantly cytoplasmic staining, consistent with HPA heart and skeletal-muscle IHC and UniProt localisation (HPA tissue IHC; UniProt P50461 subcellular). Limited nuclear signal may merit separate review because CSRP3 shuttles between compartments, while HPA labels additional nucleoplasmic localisation uncertain (UniProt P50461 subcellular; HPA subcellular). Be cautious with signal in unrelated cells, at section edges or folds, or in necrotic regions; compare morphology and no-primary controls before assigning specificity (HPA tissue IHC; standard IHC practice). If a peroxidase-based stain persists without primary antibody, investigate endogenous enzyme activity and blocking before interpreting the DAB deposit as CSRP3 (standard IHC practice).
Boster reagents
Best CSRP3 / Cysteine and glycine-rich protein 3 IHC Antibodies
A06522 has real IHC data from paraffin-embedded mouse heart (catalog IHC image caption). Its listed reactivity is Human, Mouse and Rat (catalog reactivity); no IF image is supplied (catalog image fields).
Real IHC data
Anti-CSRP3 Antibody
Cat # A06522
A06522 is listed for IHC and WB, with Human, Mouse and Rat reactivity (catalog applications and reactivity). Its IHC image shows paraffin-embedded mouse heart stained at 1:100 after microwave retrieval in 10 mM PBS, pH 7.2 (catalog IHC image caption).
Which to pick: Choose A06522 for paraffin-section IHC because its own image documents that preparation in mouse heart; the fixative is unreported (catalog IHC image caption). No SKU in the payload is listed for IF/ICC or has an IF image, so there is no documented IF/ICC choice here (catalog applications and image fields). A06522 is the cross-species candidate based on listed Human, Mouse and Rat reactivity, although its image documents mouse tissue only; it is rabbit-hosted and its clonality is unreported (catalog reactivity, IHC image caption, host and clone fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P50461 (CSRP3_HUMAN, Cysteine and glycine-rich protein 3).
- Human Protein Atlas. CSRP3 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CSRP3 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the nucleoplasm..
- Human Protein Atlas. CSRP3 antibody validation summary (1 antibodies).
- Integrated proteomic and transcriptomic profiling identifies aberrant gene and protein expression in the sarcomere, mitochondrial complex I, and the extracellular matrix in Warmblood horses with myofibrillar myopathy. BMC genomics 2021 — PMC8194174.
- A novel strategy for the photothermal treatment and electroencephalogram monitoring of in situ glioma. RSC advances 2026 — PMC13223896.
- PubMed PMID:7490106 — UniProt-cited evidence.
- PubMed PMID:24860983 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CSRP3 (Cysteine and glycine-rich protein 3). Expected localisation Muscle cytoplasm observed; Z line expected (HPA tissue IHC; UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A06522 at 1:50-1:200 (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: Paraffin-embedded mouse heart tissue; fixative not specified (datasheet A06522); 2. Fixation: Image fixative and duration unreported (datasheet A06522); 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 Tris-EDTA buffer, pH 9.0 , 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-CSRP3, 1:50-1:200 (datasheet A06522); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Isoform coverage is unknown without antibody epitope data (UniProt; datasheet); Isoform / epitope: Two isoforms; antibody epitope coverage is unspecified (UniProt; datasheet); Regulation: Isoform 2 is higher in muscle disease (UniProt). Evidence: UniProt P50461, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A06522, PMC13223896. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CSRP3/immunohistochemistry-ihc-guide.