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SFRP1 / Secreted frizzled-related protein 1 · IHC design guide
Design Immunohistochemistry for SFRP1
Plan chromogenic SFRP1 IHC on paraffin sections with the catalog antibody at 0.5–1 μg/mL (datasheet A01968-2). Assess extracellular staining and compare high signal in testis pachytene spermatocytes with adipocytes where staining was not detected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SFRP1 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 | Extracellular positivity in several tissues (HPA tissue IHC) | |
| Staining pattern | Extracellular signal; high in testis pachytene spermatocytes (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A01968-2) | |
| Positive control | Testis | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01968-2) | |
| Caveat | Secreted protein: staining may differ from RNA location (HPA tissue IHC) | |
| Regulation | Specific expression regulation unreported (UniProt) | |
| Isoform / epitope | No isoforms listed; map epitopes to mature chain 32–314 (UniProt) |
Section 1
Recommended SFRP1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A01968-2) is accompanied by three published SFRP1 IHC methods (PMC2361362; PMC7565692; PMC4342152).
| Sample | Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet A01968-2) |
| Fixation | Image fixative and duration unreported (datasheet A01968-2); 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 A01968-2) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01968-2) |
| Primary antibody | Rabbit anti-SFRP1, 0.5-1μg/ml (datasheet A01968-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01968-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01968-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SFRP1-positive staining in pachytene spermatocytes of testis (HPA tissue IHC: High). HPA tissue profile: extracellular positivity in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate pH 6 retrieval (datasheet: A01968-2); Tris-EGTA pH 9 is a published alternative (PMC7565692).
Section 2
What Is the Expected SFRP1 Staining Pattern?
SFRP1 is secreted and can remain at the cell surface or in extracellular matrix; it has no transmembrane segment (UniProt Q8N474: location and topology). In tissue IHC, expect extracellular positivity in several tissues and high staining associated with pachytene spermatocytes in testis (HPA: tissue IHC). HPA rates the tissue staining Approved and cautions that RNA and protein locations may differ for a secreted protein (HPA: tissue IHC).
| Extracellular staining, with high staining associated with pachytene spermatocytes in testis (HPA: tissue IHC). | This fits the reported tissue pattern (HPA: tissue IHC). Cell-associated signal is plausible because secreted SFRP1 may remain cell-surface-associated; score the stained compartment and cells separately (UniProt Q8N474: location; standard IHC practice). |
| Strong, isolated nuclear staining with no corresponding extracellular pattern (standard IHC practice). | Treat this as a discordant result requiring investigation: UniProt places SFRP1 outside the cell or at its surface and lists no transmembrane segment (UniProt Q8N474: location and topology). Check antibody controls and detection background before assigning nuclear localisation (standard IHC practice). |
| Prominent signal in adipocytes or respiratory epithelial cells (HPA: tissue IHC comparison). | HPA reports SFRP1 as not detected in adipocytes of adipose tissue and respiratory epithelial cells of bronchus (HPA: tissue IHC). In those sampled contexts, investigate cross-reactivity or endogenous detection activity; an unexpected stain alone does not establish either cause (standard IHC practice). |
| Diffuse chromogen across cells and surrounding tissue, obscuring compartment boundaries (standard IHC practice). | The pattern cannot reliably distinguish the extracellular signal HPA reports from nonspecific background (HPA: tissue IHC; standard IHC practice). Compare the no-primary control and inspect whether signal follows tissue structure before scoring SFRP1 (standard IHC practice). |
| No staining associated with pachytene spermatocytes in testis (HPA: tissue IHC comparison). | This conflicts with HPA's high staining observation for those cells, but a single negative section does not prove target absence (HPA: tissue IHC; standard IHC practice). Confirm that the relevant cells are present and review the staining controls and run conditions (standard IHC practice). |
💡Expected SFRP1 appearanceCall a result positive when staining follows an extracellular pattern and, in testis, shows high signal associated with pachytene spermatocytes; isolated nuclear staining or uniform haze is suspect (HPA: tissue IHC; UniProt Q8N474: location; standard IHC practice).
| Secretion and surface association (UniProt Q8N474: location and topology). | SFRP1 can appear outside cells, in extracellular matrix, or at the cell surface despite lacking a transmembrane segment; do not require a crisp membrane rim to recognise a plausible pattern (UniProt Q8N474: location and topology). |
| Processing and epitope position (UniProt Q8N474: processing). | The signal peptide occupies residues 1–31, while the mature chain is 32–314 (UniProt Q8N474: processing). If an antibody's epitope becomes known, compare its position with that boundary; no epitope is supplied here (UniProt Q8N474: processing; supplied antibody record). |
| Tissue context and secreted protein (HPA: tissue IHC; UniProt Q8N474: location). | HPA reports extracellular positivity in several tissues and high staining in testicular pachytene spermatocytes, while its sampled adipocytes and bronchial respiratory epithelium are not detected (HPA: tissue IHC). Compare like cells and compartments, not whole-section darkness (standard IHC practice). |
| Evidence level for the staining pattern (HPA: tissue IHC and antibody record). | The tissue IHC reliability is Approved, and CAB008116 is listed as IHC Approved (HPA: tissue IHC and antibody record). This record does not claim an Enhanced pattern or provide an ICC-IF validation result (HPA: antibody record). |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected testis signal is absent (HPA: tissue IHC comparison). | The relevant pachytene spermatocytes may be missing from the examined area, or the IHC run may have failed; the image alone cannot distinguish these possibilities (standard IHC practice). | Locate the relevant cells on the counterstained section, check the run's positive control, then review antibody dilution, retrieval and detection as general IHC variables (standard IHC practice). |
| Extracellular signal is lost in widespread haze (HPA: tissue IHC comparison). | Background from antibody binding or detection can obscure the reported extracellular pattern; no SFRP1-specific cause is established by this appearance (HPA: tissue IHC; standard IHC practice). | Inspect a no-primary control, then review blocking, washing, antibody concentration and chromogen development as general IHC variables (standard IHC practice). |
| Signal is chiefly nuclear (standard IHC practice). | That compartment disagrees with SFRP1's secreted, cell-surface or extracellular location; artefact or cross-reactivity should be considered, but is not proven (UniProt Q8N474: location; standard IHC practice). | Compare compartment-specific signal with controls and the HPA extracellular pattern before reporting nuclear SFRP1 (HPA: tissue IHC; standard IHC practice). |
| Adipocytes or bronchial respiratory epithelial cells stain strongly (HPA: tissue IHC comparison). | These sampled cell types are reported not detected by HPA; cross-reactivity or endogenous detection activity is possible, while context differences remain possible (HPA: tissue IHC; standard IHC practice). | Check the no-primary and detection controls, identify the stained cell type, and report the discrepancy with the HPA reference rather than assuming specificity (HPA: tissue IHC; standard IHC practice). |
| Staining varies between sections (standard IHC practice). | Cell composition and routine IHC processing can differ between sections; the supplied sources do not establish SFRP1-specific fixation sensitivity (standard IHC practice; HPA: tissue IHC; UniProt Q8N474). | Compare matching tissue regions and controls, then review section handling and run conditions without attributing variation to a documented SFRP1 fixation effect (standard IHC practice). |
| Can this IHC pattern establish an expected ICC-IF localisation? (HPA: subcellular and antibody records). | HPA supplies no main ICC-IF location or cell-line images, and the listed antibody has no ICC validation entry (HPA: subcellular and antibody records). | Use the separate IF/ICC guide for assay design; treat any fluorescent compartment assignment as provisional until supported by IF controls (HPA: subcellular and antibody records; standard IF practice). |
Experiment evidence
IHC & IF images showing SFRP1 staining
A01968-2 · IHCIHC analysis of SFRP1 using anti-SFRP1 antibody (A01968-2). SFRP1 was detected in paraffin-embedded section of human colon cancer tissue. He
A01968-2 · IHCIHC analysis of SFRP1 using anti-SFRP1 antibody (A01968-2). SFRP1 was detected in paraffin-embedded section of human lung cancer tissue. Hea
A01968-2 · IFIF analysis of SFRP1 using anti-SFRP1 antibody (A01968-2). SFRP1 was detected in immunocytochemical section of U20S cells. Enzyme antigen re
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000104332-SFRP1/tissue/cerebral+cortex#imid_2241216
HPA · Salivary glandHuman Protein Atlas IHC, Salivary gland. Source: https://www.proteinatlas.org/ENSG00000104332-SFRP1/tissue/salivary+gland#imid_2241164
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000104332-SFRP1/tissue/colon#imid_2237782
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000104332-SFRP1/tissue/liver#imid_2237783
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SFRP1 IHC & IF
🧪Run testis first and score pachytene spermatocytes, where staining is high (HPA: Testis, pachytene spermatocytes, High); run adrenal gland as the negative tissue and assess its glandular cells (HPA: Adrenal gland, glandular cells, Not detected). On the testis slide, use cells without discernible target staining as an internal background reference; they should show intact morphology and only background-level chromogen, without assuming every other cell type is negative (HPA: High is specified for pachytene spermatocytes).
Positive control tissue: Testis (Pachytene spermatocytes, 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 SFRP1; derive a cell-line control from the positive tissue's cell type (Pachytene spermatocytes) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; a concentration-matched rabbit IgG isotype control for the rabbit primary; and a biological specificity control using knockout tissue or immunizing-peptide competition if available (selected A01968-2 caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase and check endogenous biotin background before using the caption’s biotin-based, DAB detection system on testis (selected A01968-2 caption: biotinylated secondary, SABC and DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A01968-2 paraffin-section caption does not report a fixative (selected A01968-2 caption). That caption uses citrate retrieval at pH 6 for 20 minutes, but does not establish that retrieval is required (selected A01968-2 caption). No matched frozen-section or IF/ICC comparison is supplied, so neither can be judged easier; in testis, resolve individual germ-cell morphology and background before scoring pachytene staining (HPA: Testis, pachytene spermatocytes, High; standard IHC practice).
HPA tissue IHC evidence for SFRP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Testis | Pachytene spermatocytes | 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 SFRP1 IHC Tips
Troubleshooting SFRP1 staining in paraffin section IHC requires attention to retrieval, extracellular localisation, and controls that distinguish specific signal from tissue background.
How should I retrieve SFRP1 when staining is weak in paraffin sections?
Start with heat-mediated antigen retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A01968-2). The selected paraffin-section image used that retrieval before overnight incubation with 1 μg/mL catalog antibody at 4°C (datasheet A01968-2). If staining is weak, first check section adherence, heating consistency, and reagent performance while keeping retrieval conditions fixed (standard IHC practice). Then compare a modestly adjusted heating time on matched sections, recording tissue damage alongside signal (standard IHC practice). An alternative buffer should be a documented fallback experiment, since the supplied product evidence supports citrate at pH 6 (datasheet A01968-2).
Could fixation explain weak or uneven SFRP1 staining?
The selected paraffin-section caption does not state a fixative, so target-specific fixation sensitivity is unknown (datasheet A01968-2). Record the actual fixative and fixation duration for each specimen before attributing weak staining to SFRP1 biology (standard IHC practice). Compare matched sections with a consistent citrate pH 6 retrieval step and the same antibody incubation; this isolates processing differences more clearly (datasheet A01968-2; standard IHC practice). Examine morphology and section integrity alongside staining, because harsh processing can complicate interpretation (standard IHC practice). Do not infer fixation tolerance from the reported extracellular tissue pattern or from protein topology (HPA tissue IHC; UniProt Q8N474 topology).
Should SFRP1 appear inside cells or in extracellular tissue?
SFRP1 is secreted and can associate with the cell membrane or extracellular matrix; it has no transmembrane segment (UniProt Q8N474 subcellular location and topology). Accordingly, assess extracellular and pericellular chromogenic signal alongside any cell-associated staining (UniProt Q8N474 subcellular location; standard IHC practice). The tissue atlas reports extracellular positivity in several tissues, while its protein and RNA locations may differ for a secreted variant (HPA tissue IHC). Use matched negative controls and inspect the same anatomical regions before assigning a cellular source to deposited protein (standard IHC practice). A sharply intracellular pattern alone deserves further validation, especially if extracellular structures remain unstained (UniProt Q8N474 subcellular location; standard IHC practice).
How can epitope location affect SFRP1 staining in sections?
The supplied record lists 0 isoforms and a signal peptide at residues 1–31, followed by the secreted chain at 32–314 (UniProt Q8N474 processing and isoforms). Its FZ domain spans 53–169, its NTR domain 186–306, and it has a glycosylation site at 173 (UniProt Q8N474 domains and glycosylation). The catalog antibody epitope is not specified in the supplied evidence (datasheet A01968-2). Therefore, do not infer whether retrieval exposes its epitope or whether staining detects precursor versus secreted protein (UniProt Q8N474 processing; standard IHC practice). If staining differs across compartments, compare controls and tissue morphology before assigning that difference to an epitope or processing state (standard IHC practice).
How should I investigate SFRP1 by multiplex immunofluorescence?
Treat IF as a separate application: the supplied product image documents chromogenic staining in a paraffin section, not IF performance (datasheet A01968-2). In testis, pair SFRP1 with a validated marker for pachytene spermatocytes, the reported high-staining cell population, while also inspecting nearby extracellular signal (HPA tissue IHC; UniProt Q8N474 subcellular location). Choose spectrally separated fluorophores and consider a far-red channel if tissue autofluorescence obscures the target channel (standard IF practice). Because SFRP1 is secreted without a transmembrane segment, mild or no permeabilisation may suit extracellular epitopes; an intracellular precursor epitope would require a separately validated permeabilisation condition (UniProt Q8N474 topology and processing; standard IF practice). Verify each channel with single-stain and omission controls (standard IF practice).
What should I check when SFRP1 DAB staining is diffuse?
Separate plausible extracellular SFRP1 signal from general brown haze by comparing anatomical distribution, morphology, and matched negative controls (UniProt Q8N474 subcellular location; standard IHC practice). The selected image used a 10% goat serum block, biotinylated secondary antibody, avidin-biotin detection, and DAB (datasheet A01968-2). Include secondary-only controls and check endogenous biotin where avidin-biotin detection is used (standard IHC practice). Apply an appropriate endogenous peroxidase block and monitor DAB development consistently across sections (standard IHC practice). If staining concentrates at folds, torn edges, or necrotic areas, assess intact regions before calling the diffuse signal positive (standard IHC practice).
How should I score extracellular SFRP1 staining in paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define tissue regions and exclusion criteria before scoring, and keep retrieval, DAB development, and image acquisition consistent across specimens (standard IHC practice). For cell-associated staining, record the percentage of positive cells and intensity, then calculate an H-score only when cellular boundaries can be scored reliably (standard IHC practice). For diffuse extracellular signal, measure positive area or optical density per mm² of viable tissue instead of forcing a cell-based score (UniProt Q8N474 subcellular location; standard IHC practice). Normalise to viable tissue area or the relevant anatomical compartment, and exclude folds and necrosis using the same rules throughout (standard IHC practice). Report extracellular and cell-associated measurements separately because deposited secreted protein need not identify its producing cell (UniProt Q8N474 subcellular location).
When is an apparent SFRP1-positive region likely to be artefactual?
Interpret convincing signal in light of SFRP1 secretion and its reported extracellular or matrix-associated localisation (UniProt Q8N474 subcellular location). The tissue atlas reports high staining in testis pachytene spermatocytes and no detected staining in the listed adipocytes and several other specified cell populations; these are comparison points, not universal controls (HPA tissue IHC). Question isolated nuclear staining or a pattern confined to section edges, folds, or necrotic tissue, and inspect adjacent intact regions (UniProt Q8N474 subcellular location; standard IHC practice). Test suspected endogenous enzyme or detection-system signal with appropriate reagent-omission controls (standard IHC practice). Because secreted protein may accumulate away from its producing cells, avoid equating a DAB-positive deposit with local SFRP1 expression (UniProt Q8N474 subcellular location; HPA tissue IHC).
Boster reagents
Best SFRP1 / Secreted frizzled-related protein 1 IHC Antibodies
Anti-SFRP1 image captions show paraffin-section IHC in human colon and lung cancers and mouse intestine and kidney, plus IF/ICC in U20S cells (catalog image captions).
Real IHC data
Anti-SFRP1 Antibody ®
Cat # A01968-2
A01968-2 will render with its own human colon cancer paraffin-section IHC figure; other captions show human lung cancer and mouse intestine and kidney IHC, plus U20S cell IF/ICC (catalog image captions). M01968 is listed for human IHC and ICC/IF but has no IHC or IF image caption, so it has no figure card here (catalog applications, reactivity and image alts).
Which to pick: Choose A01968-2 for tissue IHC: its pictured human colon cancer paraffin section used citrate retrieval at pH 6 for 20 minutes and 1 μg/ml primary antibody; the fixative is unreported (A01968-2 IHC image caption). For IF/ICC, A01968-2 has a pictured U20S cell example at 2 μg/ml, while M01968 lists ICC/IF for human samples without a supplied image (A01968-2 IF image caption; M01968 catalog applications, reactivity and image alts). Choose A01968-2 for cross-species work because it lists human, mouse and rat reactivity and has pictured human and mouse paraffin-section IHC; M01968 is a rabbit monoclonal listed for human reactivity only (catalog reactivity and clone; A01968-2 IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8N474 (SFRP1_HUMAN, Secreted frizzled-related protein 1).
- Human Protein Atlas. SFRP1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. SFRP1 subcellular location (ICC-IF): Variation in transcript expression is not correlated to the cell cycle..
- Human Protein Atlas. SFRP1 antibody validation summary (1 antibodies).
- The Wnt antagonist sFRP1 is downregulated in premalignant large bowel adenomas. British journal of cancer 2006 — PMC2361362.
- Secreted Frizzled-Related Protein 1 as a Biomarker against Incomplete Age-Related Lobular Involution and Microcalcifications' Development. Cancers 2020 — PMC7565692.
- Diagnostic value of SFRP1 as a favorable predictive and prognostic biomarker in patients with prostate cancer. PloS one 2015 — PMC4342152.
- Deregulation of secreted frizzled-related proteins is associated with aberrant β-catenin activation in the carcinogenesis of oral submucous fibrosis. OncoTargets and therapy 2015 — PMC4610781.
- PubMed PMID:9192640 — UniProt-cited evidence.
- PubMed PMID:9391078 — UniProt-cited evidence.
- PubMed PMID:9724099 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SFRP1 (Secreted frizzled-related protein 1). Expected localisation Extracellular positivity in several tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A01968-2). Validated IHC antibody: A01968-2 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Pachytene spermatocytes); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet A01968-2); 2. Fixation: Image fixative and duration unreported (datasheet A01968-2); 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 A01968-2); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01968-2); 8. Primary antibody: Rabbit anti-SFRP1, 0.5-1μg/ml (datasheet A01968-2); 9. Primary incubation: Overnight at 4 °C (datasheet A01968-2). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01968-2); Caveat: Secreted protein: staining may differ from RNA location (HPA tissue IHC); Isoform / epitope: No isoforms listed; map epitopes to mature chain 32–314 (UniProt); Regulation: Specific expression regulation unreported (UniProt). Evidence: UniProt Q8N474, HPA tissue IHC · Approved, HPA cell-line RNA · Variation in transcript expression is not correlated to the cell…, Datasheet A01968-2, PMC2361362, PMC7565692. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SFRP1/immunohistochemistry-ihc-guide.