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C1QTNF6 / Complement C1q tumor necrosis factor-related protein 6 · IHC design guide
Design Immunohistochemistry for C1QTNF6
Start C1QTNF6 chromogenic IHC-P with the catalog antibody at 10 μg/mL (datasheet: A12414-1). Compare placental or endometrial staining with an HPA tissue showing no detected signal, and interpret the cytoplasmic pattern in light of the protein’s secretion (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
C1QTNF6 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 | Cytoplasmic in trophoblasts and endometrial stroma (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic trophoblast and endometrial stromal staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Placenta+1 more · see all | |
| 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 A12414-1) | |
| Caveat | Secretion may separate tissue RNA and protein patterns (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | 3 isoforms; signal 1–46 is cleaved; epitope map unknown (UniProt) |
Section 1
Recommended C1QTNF6 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by four published C1QTNF6 IHC workflows for bladder, lung and oral cancer specimens (PMC7585664; PMC7805025; PMC8670214; PMC9310408).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A12414-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-C1QTNF6, 10 μg/mL (datasheet A12414-1) |
| 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 | C1QTNF6-positive staining in endothelial cells of placenta (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in placental trophoblasts and endometrial stroma. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval); a published lung protocol uses a 3 min citrate boil (PMC9310408).
Section 2
What Is the Expected C1QTNF6 Staining Pattern?
C1QTNF6 is secreted and has no transmembrane segment (UniProt Q9BXI9 topology). In tissue IHC, expect cytoplasmic staining in placental trophoblasts and endometrial stromal cells (HPA: tissue profile). HPA also records high staining in placental endothelial cells and medium staining in endometrial stromal cells (HPA: tissue IHC). Its tissue IHC reliability is Enhanced, with a caveat that secretion can separate the locations of RNA and protein (HPA: reliability description).
| Cytoplasmic staining appears in placental trophoblasts or endometrial stromal cells. | This matches the reported tissue pattern (HPA: tissue profile). Compare cell identity and signal with a negative control before scoring; a cytoplasmic stain alone does not establish specificity (general IHC practice). |
| Placental endothelial cells stain strongly; endometrial stromal cells stain less strongly. | HPA reports High staining in placental endothelial cells and Medium staining in endometrial stromal cells (HPA: tissue IHC). These are observations in different tissue and cell contexts, not a required intensity ratio for every specimen (HPA: tissue IHC). |
| Prominent nuclear staining dominates a paraffin tissue section. | Treat this as discordant with the reported cytoplasmic tissue pattern and investigate artefact or off-target staining (HPA: tissue profile; general IHC practice). HPA separately reports approved nucleoplasmic ICC-IF localisation; that observation does not redefine the expected tissue IHC pattern (HPA: subcellular ICC-IF). |
| Unexpected cell types stain, including adipocytes in adipose tissue. | HPA reports adipocytes as Not detected in adipose tissue (HPA: tissue IHC). Check for cross-reactivity or endogenous detection activity with an appropriate negative control; neither cause can be assigned from appearance alone (general IHC practice). |
| Colour is diffuse across tissue, or a known-positive compartment has no signal. | Diffuse colour limits cell-specific scoring and calls for a background control (general IHC practice). If placental endothelial cells or endometrial stromal cells are blank, first verify that the staining run worked before interpreting a biological absence (HPA: tissue IHC; general IHC practice). |
💡Expected C1QTNF6 appearanceCall the IHC result positive when interpretable cytoplasmic staining occurs in the reported placental or endometrial cell populations, with High placental endothelial and Medium endometrial stromal staining as HPA reference observations; widespread nuclear or cell-independent colour is suspect (HPA: tissue IHC and tissue profile; general IHC practice).
| Secreted protein and tissue context | C1QTNF6 has a signal peptide at residues 1–46 and a chain at 47–278, with no transmembrane segment (UniProt Q9BXI9 topology and processing). Secreted protein can make protein staining diverge from local RNA expression; HPA explicitly flags this limitation (HPA: reliability description). |
| Structural features and epitope uncertainty | UniProt lists a glycosylation site at residue 91, a collagen-like region at 97–138, and a C1q domain at 139–259 (UniProt Q9BXI9). The supplied record gives no antibody epitope, so these features cannot determine an antigen retrieval condition or predict loss of staining after fixation. |
| Isoforms and antibody validation | UniProt lists three isoforms (UniProt Q9BXI9). HPA lists HPA002042 as a rabbit polyclonal antibody with Enhanced IHC validation (HPA: antibodies). Without an epitope map or isoform-specific staining data, do not infer which isoform accounts for a positive cell. |
| IF/ICC Q&A: Should the IF result be judged by the tissue IHC pattern? | No. HPA reports approved nucleoplasmic localisation in ICC-IF and lists images from A-431, U-251MG, and U2OS (HPA: subcellular ICC-IF). Use that as an assay-specific observation; the reported tissue IHC pattern is cytoplasmic (HPA: tissue profile). |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in a placental or endometrial section. | A run failure or weak detection is possible; HPA reports positive placental endothelial and endometrial stromal staining (HPA: tissue IHC). | Check a known-positive section in the same run, then review antibody dilution, retrieval, detection reagents, and counterstain as general IHC controls (general IHC practice). No target-specific retrieval condition is supplied. |
| The whole section has diffuse chromogenic colour. | Nonspecific binding or background from detection reagents can obscure cell-specific staining (general IHC practice). | Compare a no-primary control, review blocking and washes, and adjust detection or antibody concentration if the controls identify background (general IHC practice). Score only interpretable cellular signal. |
| Strong nuclear colour appears in tissue IHC. | This conflicts with HPA's cytoplasmic tissue profile; nucleoplasmic ICC-IF approval belongs to a separate assay (HPA: tissue profile; HPA: subcellular ICC-IF). | Check the negative control and staining distribution, then confirm cell identity before treating nuclear tissue colour as C1QTNF6 (general IHC practice). |
| Adipocytes or other unexpected cells stain. | Adipocytes are Not detected in HPA adipose tissue IHC; cross-reactivity or endogenous detection activity is possible (HPA: tissue IHC; general IHC practice). | Review a no-primary control and compare the suspect cells with a reported positive population; investigate the detection system if the control also stains (general IHC practice). |
| Placental and endometrial cell intensities differ. | HPA records High placental endothelial staining and Medium endometrial stromal staining, while secretion complicates correspondence between RNA and protein location (HPA: tissue IHC and reliability description). | Score each identified cell population against its own tissue context and the run controls; do not require equal intensity across the two tissues (HPA: tissue IHC; general IHC practice). |
| The investigator wants to change retrieval because the target is glycosylated. | The listed glycosylation site does not establish fixation sensitivity or a preferred retrieval method (UniProt Q9BXI9; supplied evidence scope). | Use a validated IHC-P condition if available and evaluate any retrieval change against positive and negative controls (general IHC practice). Target-specific fixation sensitivity is unreported in the supplied sources. |
Experiment evidence
IHC & IF images showing C1QTNF6 staining
A12414-1 · IHCImmunohistochemistry of CTRP6 in human brain tissue with CTRP6 antibody at 10 μg/mL.
A12414-1 · IFImmunofluorescence of CTRP6 in Human Brain cells with CTRP6 antibody at 20 μg/mL.
A12414 · IFImmunocytochemistry of CTRP6 in HeLa cells with CTRP6 antibody at 10 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000133466-C1QTNF6/tissue/cerebral+cortex#imid_1064648
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000133466-C1QTNF6/tissue/colon#imid_1064566
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000133466-C1QTNF6/tissue/liver#imid_1064575
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000133466-C1QTNF6/tissue/kidney#imid_1064576
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA002042; green = C1QTNF6, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA002042; green = C1QTNF6, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for C1QTNF6 IHC & IF
🧪Run placenta first and score endothelial cells, where C1QTNF6 staining is High (HPA: High in placental endothelial cells). Use adipose tissue as a negative comparison, scoring adipocytes reported as Not detected (HPA: Not detected in adipocytes); neighboring cells without specific staining on the placental slide should show only background signal as an internal reference (standard IHC practice).
Positive control tissue: Placenta (Endothelial cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show C1QTNF6 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and clonality-matched isotype controls, plus a knockout sample or peptide-block control where available (standard IHC practice). Block endogenous peroxidase in placental sections and, if using avidin–biotin detection, assess endogenous biotin staining (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; the selected A12414-1 tissue-IHC caption also does not report a fixative (selected SKU caption: fixative not stated). The evidence does not establish whether frozen sections or IF are easier for this target (supplied application evidence). Because placental endogenous peroxidase or biotin can contribute background, check the corresponding detection controls before scoring endothelial staining (standard IHC practice).
HPA tissue IHC evidence for C1QTNF6
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. 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 |
|---|---|---|---|---|
| Placenta | Endothelial cells | High | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Endocrine 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 C1QTNF6 IHC Tips
Troubleshoot CTRP6 staining in paraffin section IHC, with a separate note on IF multiplexing and interpretation.
What retrieval conditions should I start with for CTRP6 in paraffin sections?
Start with citrate buffer at pH 6.0 for heat induced retrieval at 95–98 °C for 20 min (page retrieval rule). Let sections cool in the retrieval buffer, then compare staining and tissue preservation across identically processed sections (standard IHC practice). If staining remains weak, test a different buffer or retrieval duration on adjacent sections while keeping antibody concentration and detection conditions fixed (standard IHC practice). Score the expected cell populations separately from diffuse extracellular signal, since CTRP6 is secreted and staining need not remain in its cell of origin (UniProt Q9BXI9; HPA tissue IHC).
How should I troubleshoot weak CTRP6 staining when fixation history is uncertain?
Target specific sensitivity to fixation is unknown from the supplied evidence, so record the fixative and fixation interval for each specimen (supplied evidence; standard IHC practice). The catalog antibody’s human brain tissue IHC caption reports 10 µg/mL antibody but does not state the fixative (catalog antibody A12414-1 caption). Compare sections with documented processing alongside a positive tissue control, applying the same pH 6.0 retrieval and detection workflow to both (page retrieval rule; standard IHC practice). If signal differs, assess tissue preservation and control staining before attributing the difference to CTRP6 abundance or a fixation effect (standard IHC practice).
Where should CTRP6 staining appear, and how should conflicting compartments be assessed?
CTRP6 is a secreted protein with no transmembrane segment, so extracellular signal may accompany staining in producing cells (UniProt Q9BXI9). HPA tissue IHC describes cytoplasmic expression in placental trophoblasts and endometrial stroma, while its subcellular IF record assigns an approved nucleoplasmic location (HPA tissue IHC; HPA subcellular). Score cytoplasmic, nuclear and extracellular staining separately rather than combining them into one positive fraction (standard IHC practice). If a compartment dominates unexpectedly, inspect matched negative controls and the relevant cell population, then seek independent antibody or orthogonal evidence before assigning that pattern to CTRP6 (standard IHC practice).
Could isoforms or epitope processing explain different CTRP6 staining patterns?
The record lists 3 isoforms and a signal peptide at residues 1–46; the annotated chain spans residues 47–278 (UniProt Q9BXI9). It also places one glycosylation site at residue 91, a collagen like region at 97–138 and a C1q domain at 139–259 (UniProt Q9BXI9). Check the antibody’s stated immunogen or epitope against those features before proposing an isoform or processing explanation for discrepant staining (standard IHC practice). Without an epitope map and isoform specific validation, differences between specimens cannot establish which CTRP6 isoform produced the chromogenic signal (standard IHC practice; UniProt Q9BXI9).
How can IF help verify the cells and compartment labelled by CTRP6 IHC?
Pair CTRP6 with a marker identifying the expected cell population, and evaluate overlap at the single cell level rather than comparing whole fields (HPA tissue IHC; standard IF practice). For the reported placental trophoblast or endometrial stromal patterns, choose a cell marker validated in the specimen and include single stain controls for each channel (HPA tissue IHC; standard IF practice). Select fluorophores after checking tissue autofluorescence in an unstained section, placing weaker specific signal in a cleaner spectral channel (standard IF practice). CTRP6 has no transmembrane segment, so choose permeabilisation according to the antibody’s mapped epitope and the intracellular compartment being tested (UniProt Q9BXI9; standard IF practice).
How do I distinguish diffuse CTRP6 signal from chromogenic IHC background?
Run a no primary control and inspect whether diffuse color persists in the same regions, particularly around damaged tissue or section edges (standard IHC practice). For peroxidase based detection, verify the endogenous peroxidase block and assess whether residual enzyme activity contributes to DAB deposition (standard IHC practice). Titrate the antibody and detection exposure on adjacent sections while retaining a positive control, since stronger development can obscure cell boundaries (standard IHC practice). Because CTRP6 is secreted, extracellular staining is plausible, but accept it as specific only when its distribution is reproducible and controls exclude staining caused by the workflow (UniProt Q9BXI9; standard IHC practice).
What should I measure when CTRP6 staining includes cells and extracellular space? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then report the percentage of positive cells and an H-score for cell associated chromogenic staining (standard IHC practice). If the scientific question concerns extracellular deposits, measure stained area or signal density per mm² of viable tissue as a separate endpoint (standard IHC practice). Normalize cell counts to the number of eligible cells and area measurements to the analyzable tissue area, excluding folds, necrosis and blank space (standard IHC practice). Keep retrieval, antibody concentration, DAB development and image thresholds consistent across specimens, and interpret scores within the specified tissue compartment (standard IHC practice; page retrieval rule).
When is a CTRP6 positive IHC result convincing rather than artefactual?
A convincing result follows a reproducible cell and compartment pattern with low signal in matched negative controls (standard IHC practice). HPA reports cytoplasmic staining in placental trophoblasts and endometrial stroma, but also an approved nucleoplasmic IF location, so compartment disagreement needs independent validation (HPA tissue IHC; HPA subcellular). Inspect staining at cut edges, necrotic regions and sites of residual endogenous enzyme activity before scoring an apparent positive field (standard IHC practice). The catalog antibody’s human brain caption demonstrates tissue IHC staining at 10 µg/mL without reporting fixation; it does not establish that every brain cell type should stain (catalog antibody A12414-1 caption).
Boster reagents
Best C1QTNF6 / Complement C1q tumor necrosis factor-related protein 6 IHC Antibodies
The catalog includes a paraffin-section IHC antibody with a human brain tissue image (A12414-1 image caption) and IF/ICC antibodies with human cell images (catalog image captions). Listed reactivity spans human, mouse and rat (catalog reactivity).
Real IHC data
Anti-CTRP6 C1QTNF6 Antibody
Cat # A12414-1
Real IF data
Anti-CTRP6 C1QTNF6 Antibody
Cat # A12414
A12414-1 lists IHC-P and IF and shows IHC in human brain tissue and IF in human brain cells (catalog applications; A12414-1 image captions). A12414 lists IF and ICC and shows both in HeLa cells (catalog applications; A12414 image captions).
Which to pick: Choose A12414-1 for paraffin-section IHC: it lists IHC-P and shows staining in human brain tissue at 10 μg/mL; the fixative is unreported (catalog applications; A12414-1 IHC image caption). Choose A12414 for cell IF/ICC, supported by its HeLa cell images; for the broadest listed species reactivity, choose A12414-1, which lists human, mouse and rat, while A12414 lists human and mouse (A12414 image captions; catalog reactivity). Both antibodies list rabbit as host, and clonality is unreported (catalog host and clone fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9BXI9 (C1QT6_HUMAN, Complement C1q tumor necrosis factor-related protein 6).
- Human Protein Atlas. C1QTNF6 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. C1QTNF6 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. C1QTNF6 antibody validation summary (1 antibodies).
- C1QTNF6 Overexpression Acts as a Predictor of Poor Prognosis in Bladder Cancer Patients. BioMed research international 2020 — PMC7585664.
- C1QTNF6 regulates cell proliferation and apoptosis of NSCLC in vitro and in vivo. Bioscience reports 2021 — PMC7805025.
- C1QTNF6 promotes oral squamous cell carcinoma by enhancing proliferation and inhibiting apoptosis. Cancer cell international 2021 — PMC8670214.
- C1QTNF6 regulated by miR-29a-3p promotes proliferation and migration in stage I lung adenocarcinoma. BMC pulmonary medicine 2022 — PMC9310408.
- PubMed PMID:12975309 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:10591208 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for C1QTNF6 (Complement C1q tumor necrosis factor-related protein 6). Expected localisation Cytoplasmic in trophoblasts and endometrial stroma (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A12414-1 at 10 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Placenta (Endothelial cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A12414-1); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-C1QTNF6, 10 μg/mL (datasheet A12414-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A12414-1); Caveat: Secretion may separate tissue RNA and protein patterns (HPA tissue IHC); Isoform / epitope: 3 isoforms; signal 1–46 is cleaved; epitope map unknown (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt Q9BXI9, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm., Datasheet A12414-1, PMC7585664, PMC7805025. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/C1QTNF6/immunohistochemistry-ihc-guide.