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CTHRC1 / Collagen triple helix repeat-containing protein 1 · IHC design guide
Design Immunohistochemistry for CTHRC1
Plan chromogenic CTHRC1 IHC on paraffin sections with the catalog antibody at 0.5–1 μg/mL (datasheet A05203-1). Compare cytoplasmic staining in intestinal glandular cells with a negative control such as prostate glandular cells (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CTHRC1 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 | Observed cytoplasm (HPA tissue IHC); secreted ECM protein (UniProt) | |
| Staining pattern | Cytoplasmic staining in gut glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05203-1) | |
| 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 sections. (standard IHC practice; not target-specific) | |
| Caveat | Secretion can separate protein and RNA tissue patterns (HPA tissue IHC) | |
| Regulation | Regulation not specified (UniProt) | |
| Isoform / epitope | 3 isoforms; mature chain 31–243; epitopes unreported (UniProt) |
Section 1
Recommended CTHRC1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: A05203-1) with two published CTHRC1 paraffin-section protocols (PMC3466254; PMC13235751).
| Sample | Paraffin-embedded human mammary cancer tissue; fixative not specified (datasheet A05203-1) |
| Fixation | Image fixative and duration unreported (datasheet A05203-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A05203-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05203-1) |
| Primary antibody | Rabbit anti-CTHRC1, 0.5-1μg/ml (datasheet A05203-1) |
| Primary incubation | Overnight at 4 °C (datasheet A05203-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A05203-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CTHRC1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet: A05203-1). Citrate pH 6.0 was reported with a different antibody (PMC3466254).
Section 2
What Is the Expected CTHRC1 Staining Pattern?
CTHRC1 is secreted into the extracellular space and matrix and has no transmembrane segment (UniProt Q96CG8 topology). In paraffin sections, expect cytoplasmic staining in several tissues, including glandular cells of the colon and trophoblastic cells of the placenta (HPA tissue IHC: High). Treat that pattern as a guide: HPA rates its tissue IHC profile Approved, pending external verification, and notes that secreted protein may not coincide with its site of RNA expression (HPA tissue IHC: reliability description).
| Strong cytoplasmic signal in colon glandular cells or placental trophoblastic cells (HPA tissue IHC: High). | This matches the reported tissue IHC pattern (HPA tissue IHC: cytoplasmic expression). Score the named cell population and its compartment; secretion means nearby extracellular signal may also be biologically plausible, but HPA does not establish it as the required positive pattern (UniProt Q96CG8: extracellular space and matrix; HPA tissue IHC: profile). |
| Predominantly nuclear staining in a paraffin tissue section, without the expected cytoplasmic pattern (HPA tissue IHC: profile). | Treat it as an unresolved IHC localisation discrepancy and check controls and detection artefacts (general IHC practice). HPA separately reports approved nucleoplasmic ICC-IF localisation, so a nuclear image alone cannot prove that an antibody is cross-reactive (HPA subcellular ICC-IF: Nucleoplasm approved). |
| Strong staining in adipocytes or prostate glandular cells, reported as Not detected by HPA (HPA tissue IHC). | Investigate cross-reactivity or endogenous detection activity, especially if a no-primary control also stains (general IHC practice). HPA's Not detected designation describes its sampled cells and assay; it does not prove universal absence across specimens or conditions (HPA tissue IHC: Not detected). |
| Widespread, similar colour across cells and empty tissue spaces, with little cell-type contrast (general IHC practice). | Diffuse background makes compartment and cell-type scoring unreliable (general IHC practice). Compare a no-primary control and review blocking, washing and chromogen development before calling the pattern CTHRC1-specific (general IHC practice); HPA reports a selective cytoplasmic tissue profile (HPA tissue IHC). |
| No signal in colon glandular cells or kidney tubule cells (HPA tissue IHC: High). | First suspect an assay or section-level failure and check an on-run positive control, antibody dilution, retrieval and detection steps (general IHC practice). A single negative section cannot overturn the HPA reference pattern, whose Approved reliability remains pending external verification (HPA tissue IHC: reliability description). |
💡Expected CTHRC1 appearanceCall a positive IHC result when the relevant cells show clear cytoplasmic staining, potentially High in colon glandular cells or placental trophoblastic cells (HPA tissue IHC: High); uniform background or staining confined to cells reported Not detected is a result to investigate, not a stand-alone positive call (HPA tissue IHC; general IHC practice).
| Secretion and topology (UniProt Q96CG8: extracellular space, matrix; no transmembrane segment). | Protein can be present beyond its producing cell, so interpret adjacent extracellular colour in tissue context (UniProt Q96CG8: localisation; general IHC practice). HPA's observed tissue IHC pattern is cytoplasmic and does not establish a required matrix-staining threshold (HPA tissue IHC: profile). |
| Maturation and glycosylation (UniProt Q96CG8: signal peptide 1–30; chain 31–243; site 186). | The annotated mature chain excludes the signal peptide (UniProt Q96CG8: processing). Epitope placement could matter for antibody recognition, but the supplied record gives no antibody epitope; do not predict staining loss or a retrieval requirement from these annotations alone (UniProt Q96CG8; general IHC practice). |
| Three annotated isoforms (UniProt Q96CG8: isoforms 1, 2 and 3). | Antibody coverage across isoforms cannot be inferred without an epitope or isoform-validation record (UniProt Q96CG8: isoforms; general IHC practice). The reported expression of isoform 1 in calcified atherosclerotic plaque and chondrocyte-like cells is not an HPA tissue IHC intensity claim (UniProt Q96CG8: tissue specificity). |
| Choice of tissue controls (HPA tissue IHC: High and Not detected categories). | Colon glandular cells and kidney tubule cells offer reported High reference patterns; adipocytes and prostate glandular cells offer reported Not detected comparisons (HPA tissue IHC). Interpret each against its named cell type, since a whole-section positive or negative label can hide mixed tissue populations (general IHC practice). |
| Antibody validation (HPA antibodies: HPA059806 IHC Approved; HPA061896 ICC Approved). | The listed validations apply to different assays and antibodies (HPA antibodies). HPA's tissue IHC reliability is Approved and pending external verification; the supplied record does not label the IHC pattern Enhanced (HPA tissue IHC: reliability description; HPA antibodies). |
| IF/ICC Q&A: Should nucleoplasmic ICC-IF staining define the paraffin IHC result? (HPA subcellular ICC-IF). | No. HPA reports approved nucleoplasmic localisation in ICC-IF, while its tissue IHC profile is cytoplasmic (HPA subcellular ICC-IF; HPA tissue IHC). Assess each assay against its own reported pattern and controls (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected glandular or trophoblastic cells are blank (HPA tissue IHC: High). | A missed assay step, ineffective detection or an unsuitable working dilution can suppress signal (general IHC practice). | Check the on-run positive section, antibody preparation and detection reagents; optimise retrieval and dilution using standard IHC controls before interpreting a biological negative (general IHC practice). No CTHRC1-specific retrieval condition or dilution is supplied. |
| A no-primary section stains alongside the test section (general IHC practice). | Endogenous enzyme activity or nonspecific detection chemistry may contribute colour in chromogenic IHC (general IHC practice). | Review the detection system and its appropriate endogenous-activity block; compare the control and test sections at the same cell types (general IHC practice). Do not assign that shared colour to CTHRC1. |
| All tissue regions show a diffuse haze that obscures the cytoplasmic pattern (HPA tissue IHC: profile). | Excess reagent, insufficient washing or overdevelopment can raise background (general IHC practice). | Review antibody dilution, blocking, washes and chromogen development with matched controls; rescore only where named cells remain distinguishable (general IHC practice). |
| Signal appears mainly nuclear in tissue IHC (HPA tissue IHC: cytoplasmic profile). | The paraffin IHC result conflicts with HPA's tissue pattern; HPA's separate ICC-IF nucleoplasmic finding does not identify the cause (HPA tissue IHC; HPA subcellular ICC-IF). | Check a no-primary control and a reported High tissue, then review antibody identity and detection settings before assigning localisation (general IHC practice; HPA tissue IHC: High). |
| Cells reported Not detected stain as strongly as the positive control (HPA tissue IHC). | Cross-reactivity or endogenous detection activity is possible, although specimens may differ from HPA's sampled material (general IHC practice; HPA tissue IHC: Not detected). | Compare the named cell populations on matched sections, inspect the no-primary control and confirm that the positive-control pattern is cytoplasmic (general IHC practice; HPA tissue IHC: profile). |
| Extracellular deposits stain while the expected cells stain weakly (UniProt Q96CG8: extracellular localisation; HPA tissue IHC: profile). | Secretion could contribute extracellular signal, but HPA's tissue profile does not validate a matrix-only positive call (UniProt Q96CG8; HPA tissue IHC). | Record cellular and extracellular staining separately, inspect background controls and compare a reported High tissue before judging specificity (general IHC practice; HPA tissue IHC: High). |
Experiment evidence
IHC & IF images showing CTHRC1 staining
A05203-1 · IHCIHC analysis of CTHRC1 using anti-CTHRC1 antibody (A05203-1). CTHRC1 was detected in paraffin-embedded section of human mammary cancer tissu
A05203-1 · IFIF analysis of CTHRC1 using anti-CTHRC1 antibody (A05203-1). CTHRC1 was detected in immunocytochemical section of HEPA1-6 cells. Enzyme anti
A05203 · IHCImmunohistochemistry of CTHRC1 in human small intestine tissue with CTHRC1 antibody at 5 μg/mL.
A05203 · IFImmunofluorescence of CTHRC1 in human small intestine tissue with CTHRC1 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000164932-CTHRC1/tissue/cerebral+cortex#imid_17400479
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000164932-CTHRC1/tissue/colon#imid_17398837
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000164932-CTHRC1/tissue/liver#imid_17398838
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000164932-CTHRC1/tissue/kidney#imid_17398839
HPA · ICC-IF PC-3Human Protein Atlas immunofluorescence, PC-3, antibody HPA061896; green = CTHRC1, blue = nucleus, red = microtubules.
HPA · ICC-IF PC-3Human Protein Atlas immunofluorescence, PC-3, antibody HPA061896; green = CTHRC1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CTHRC1 IHC & IF
🧪Run kidney first: tubule cells should stain (HPA: High in kidney cells in tubules). Use adipose tissue as a negative, with adipocytes at background (HPA: Not detected in adipocytes); on the kidney slide, assess cells outside the tubules as potential internal negatives only after confirming their background staining empirically.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CTHRC1 in PC-3, Rh30, U2OS, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit isotype controls alongside a CTHRC1 knockout specimen or validated peptide-block control (standard IHC practice; selected-SKU IHC caption: rabbit primary). For kidney, check endogenous biotin before using the caption’s streptavidin–biotin–DAB detection, and block endogenous peroxidase (standard IHC practice; selected-SKU IHC caption: biotin-based detection).
⚠️Feasibility: A CTHRC1-specific fixation window or fixation effect is unreported, and the selected A05203-1 paraffin-section caption does not state its fixative (selected-SKU IHC caption: fixative not stated). That caption used heat-mediated EDTA retrieval at pH 8.0, demonstrating a working condition without establishing that retrieval is required; kidney tubules also warrant an endogenous-biotin check with its biotin-based detection (selected-SKU IHC caption; standard IHC practice). The supplied evidence does not establish that frozen sections or IF are easier; IF localisation needs separate validation because HPA reports nucleoplasmic staining while UniProt annotates CTHRC1 as secreted and extracellular (HPA: Nucleoplasm; UniProt Q96CG8: secreted, extracellular space and matrix).
HPA tissue IHC evidence for CTHRC1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Pending external verification. At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex.). 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 | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CTHRC1 IHC Tips
Troubleshoot CTHRC1 staining in paraffin sections by checking retrieval, cellular and extracellular localisation, and controls before comparing staining scores.
What retrieval should I start with if CTHRC1 staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A05203-1). The selected paraffin-section image used that retrieval before 1 µg/mL primary antibody overnight at 4°C (datasheet A05203-1). If signal is weak, first confirm the retrieval run reached its intended temperature and that sections remained covered by buffer throughout heating (standard IHC practice). Then compare a modestly longer retrieval with the original condition on adjacent sections, keeping detection and development constant (standard IHC practice). Score both cellular and extracellular staining because CTHRC1 is secreted and associated with extracellular space and matrix (UniProt Q96CG8 localisation).
How should I troubleshoot fixation-related loss of CTHRC1 staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (datasheet A05203-1). Record each specimen’s actual fixation method and duration, then compare sections processed with the same retrieval and detection conditions (standard IHC practice). Include a section with previously reproducible staining in each run so a weak specimen can be distinguished from a failed staining run (standard IHC practice). If fixation histories differ, avoid assigning a change in signal to fixation alone without matched material (standard IHC practice). Preserve the documented EDTA pH 8.0 retrieval as the initial comparison condition (datasheet A05203-1).
Should CTHRC1 appear in cells, extracellular matrix, or nuclei?
Assess cellular and extracellular compartments separately: UniProt places CTHRC1 in extracellular space and matrix, with a signal peptide at residues 1–30 and no transmembrane segment (UniProt Q96CG8 localisation and topology). HPA reports cytoplasmic tissue staining, including high staining in placental trophoblastic cells and kidney tubule cells (HPA tissue IHC). HPA also reports an approved nucleoplasmic location in cell imaging, creating a localisation discrepancy that requires independent validation in tissue sections (HPA subcellular; HPA tissue IHC). Compare staining with tissue structure and omit-primary controls before assigning a compartment (standard IHC practice). Do not treat nuclear-only chromogenic staining as established tissue localisation solely because of the cell-imaging result (HPA subcellular).
Can the antibody distinguish CTHRC1 isoforms or a processed form?
Do not assign an isoform from staining alone: the record lists 3 isoforms but provides no antibody epitope or isoform-reactivity map (UniProt Q96CG8 isoforms; datasheet A05203-1). CTHRC1 has a signal peptide at residues 1–30 and a reported chain at 31–243, so epitope position matters when interpreting processed protein (UniProt Q96CG8 processing). Its collagen-like region spans residues 57–90, and a glycosylation site is annotated at residue 186 (UniProt Q96CG8 domains and glycosylation). Obtain the immunogen or mapped epitope before claiming recognition of every isoform or processing state (standard antibody-validation practice). Compare staining with an independent antibody or orthogonal assay when isoform specificity is central to the conclusion (standard antibody-validation practice).
How can I investigate CTHRC1 localisation by multiplex IF?
For IF, pair CTHRC1 with a marker chosen for the cell population being examined, such as a validated trophoblast marker in placenta, and inspect the channels separately before interpreting overlap (HPA tissue IHC: high in placental trophoblastic cells; standard IF practice). Select spectrally separated fluorophores and place the weaker signal in a channel with low measured tissue autofluorescence (standard IF practice). CTHRC1 has no transmembrane segment and is annotated as secreted, but this antibody’s epitope accessibility is unreported (UniProt Q96CG8 topology and localisation; datasheet A05203-1). Compare nonpermeabilised staining for accessible extracellular antigen with gently permeabilised staining for intracellular antigen, using matched acquisition settings (standard IF practice). Treat nucleoplasmic signal as a finding to validate independently because tissue IHC and cell-imaging localisation differ (HPA tissue IHC; HPA subcellular).
What should I check when CTHRC1 DAB staining is diffuse?
Begin with an omit-primary control and inspect whether colour follows section edges, damaged areas, or broad tissue surfaces (standard IHC practice). The selected image used 10% goat serum block, a biotinylated goat anti-rabbit secondary, streptavidin-biotin detection, and DAB (datasheet A05203-1). Include a peroxidase block and assess endogenous biotin when using that detection scheme; neither step establishes CTHRC1 specificity (standard IHC practice). If controls are clean but the sample remains diffuse, titrate primary antibody and shorten DAB development while keeping the EDTA pH 8.0 retrieval constant (datasheet A05203-1; standard IHC practice). Examine extracellular staining carefully because an extracellular pattern is biologically plausible for CTHRC1 (UniProt Q96CG8 localisation).
How should I score CTHRC1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the region and compartment before scoring because CTHRC1 is annotated in extracellular space and matrix, while HPA reports cytoplasmic tissue staining (UniProt Q96CG8 localisation; HPA tissue IHC). For cellular staining, record the percentage of positive cells and intensity, or calculate an H-score from percentages at intensities 0–3 (standard IHC scoring practice). For extracellular staining, report positive area or signal density per mm² of evaluable tissue, rather than forcing it into a cellular H-score (standard image-analysis practice). Normalise cellular counts to eligible cells and extracellular measurements to the measured tissue area, excluding folds and necrosis by a prespecified rule (standard image-analysis practice). Hold exposure, threshold, and DAB development criteria constant across the comparison (standard IHC quantification practice).
How do I distinguish credible CTHRC1 staining from artefact?
A credible pattern should recur in intact tissue, survive comparison with an omit-primary control, and fit a stated cellular or extracellular scoring compartment (standard IHC practice). CTHRC1 is annotated as secreted and extracellular, while HPA tissue IHC reports cytoplasmic staining; either pattern merits compartment-specific assessment (UniProt Q96CG8 localisation; HPA tissue IHC). Compare with HPA’s high placental trophoblast or kidney tubule staining and its undetected adipocyte staining as contextual references, not absolute specimen controls (HPA tissue IHC). Question signal confined to section edges, necrosis, or endogenous enzyme-rich regions when matched controls show similar colour (standard IHC practice). A nuclear-only result needs independent support because HPA’s nucleoplasmic observation comes from cell imaging rather than the tissue-IHC profile (HPA subcellular; HPA tissue IHC).
Boster reagents
Best CTHRC1 / Collagen triple helix repeat-containing protein 1 IHC Antibodies
Two anti-CTHRC1 antibodies have IHC images from human tissue and IF images from human tissue or mouse cells (catalog image captions); both list Human, Mouse and Rat reactivity (catalog applications/reactivity).
Real IHC data
Anti-CTHRC1 Antibody ®
Cat # A05203-1
Real IHC data
Anti-CTHRC1 Antibody
Cat # A05203
A05203-1 shows paraffin-section IHC in human mammary cancer tissue and IF/ICC in mouse HEPA1-6 cells (A05203-1 image captions). A05203 shows IHC and IF in human small intestine tissue (A05203 image captions).
Which to pick: Choose A05203-1 for paraffin-section IHC because its caption documents that preparation and 1 μg/mL primary antibody (A05203-1 IHC caption); the fixative is unreported (A05203-1 IHC caption). For IF/ICC in mouse cells, choose A05203-1 (A05203-1 IF caption); for IF in human small intestine tissue, choose A05203 (A05203 IF caption). Both list Human, Mouse and Rat reactivity, but their illustrated IHC results are human samples (catalog reactivity; IHC captions); clonality is unreported for both (catalog clone fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96CG8 (CTHR1_HUMAN, Collagen triple helix repeat-containing protein 1).
- Human Protein Atlas. CTHRC1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. CTHRC1 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. CTHRC1 antibody validation summary (2 antibodies).
- Adipose-derived stem cells promote the proliferation, migration, and invasion of oral squamous cell carcinoma cells by activating the Wnt/planar cell polarity signaling pathway. Translational cancer research 2022 — PMC8904952.
- Cthrc1, a novel circulating hormone regulating metabolism. PloS one 2012 — PMC3466254.
- Collagen triple helix repeat containing 1 is overexpressed in hepatocellular carcinoma and promotes cell proliferation and motility. International journal of oncology 2014 — PMC4091966.
- Microenvironmental CTHRC1 has a pro-tumorigenic role in colorectal cancer. Oncotarget 2026 — PMC13235751.
- PubMed PMID:15618538 — UniProt-cited evidence.
- PubMed PMID:12975309 — UniProt-cited evidence.
- PubMed PMID:16421571 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CTHRC1 (Collagen triple helix repeat-containing protein 1). Expected localisation Observed cytoplasm (HPA tissue IHC); secreted ECM protein (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05203-1). Validated IHC antibody: A05203-1 at 0.5-1μg/ml (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 mammary cancer tissue; fixative not specified (datasheet A05203-1); 2. Fixation: Image fixative and duration unreported (datasheet A05203-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A05203-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05203-1); 8. Primary antibody: Rabbit anti-CTHRC1, 0.5-1μg/ml (datasheet A05203-1); 9. Primary incubation: Overnight at 4 °C (datasheet A05203-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Secretion can separate protein and RNA tissue patterns (HPA tissue IHC); Isoform / epitope: 3 isoforms; mature chain 31–243; epitopes unreported (UniProt); Regulation: Regulation not specified (UniProt). Evidence: UniProt Q96CG8, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm., Datasheet A05203-1, PMC3466254, PMC13235751. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CTHRC1/immunohistochemistry-ihc-guide.