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C1QTNF6 / Complement C1q tumor necrosis factor-related protein 6 · IHC design guide
Design Immunohistochemistry for C1QTNF6
Start paraffin-section chromogenic IHC with the IHC-validated antibody at 10 μg/mL (datasheet: A12414-1). Assess cytoplasmic staining in placental trophoblasts and endometrial stroma (HPA tissue IHC), allowing for differences between protein staining and local expression (HPA tissue IHC).
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 staining in tissue (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in placental trophoblasts and endometrial stroma (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 | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Secretion can uncouple staining from local expression (HPA tissue IHC) | |
| Regulation | Enhanced in endometrium and placenta (HPA tissue RNA) | |
| Isoform / epitope | 3 isoforms; check epitope against mature chain 47–278 (UniProt) |
Section 1
Recommended C1QTNF6 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with published C1QTNF6 tissue IHC methods (PMC7585664; 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 retrieval at 95–98 °C for 20 min (page antigen-retrieval setting); adjust using the published pulmonary protocol if needed (PMC9310408).
Section 2
What Is the Expected C1QTNF6 Staining Pattern?
C1QTNF6 is a secreted protein with no transmembrane segment (UniProt Q9BXI9 topology). In paraffin-section IHC, expect cytoplasmic staining in placental trophoblasts and endometrial stromal cells (HPA tissue IHC profile). HPA reports high staining in placental endothelial cells and medium staining in endometrial stromal cells (HPA tissue IHC). Its tissue IHC reliability is Enhanced, although secretion can separate the location of protein from the cells expressing its RNA (HPA tissue IHC reliability description).
| Cytoplasmic staining in placental trophoblasts or endometrial stromal cells. | This matches the reported tissue profile (HPA tissue IHC profile). Judge each cell population separately: HPA also records high staining in placental endothelial cells and medium staining in endometrial stromal cells (HPA tissue IHC). |
| Predominantly nuclear staining in an IHC section. | This does not match HPA’s reported cytoplasmic tissue pattern (HPA tissue IHC profile). Check morphology and controls before calling it an IHC positive. Nuclear signal needs nuance: HPA separately reports approved nucleoplasmic localization by ICC-IF (HPA subcellular ICC-IF). |
| Strong staining in adipocytes or other HPA-listed negative cell populations. | Suspect nonspecific binding or endogenous detection activity and compare with appropriate controls (general IHC practice). HPA reports adipocytes as not detected in adipose tissue and breast; its negative calls apply to the listed cell populations, not every cell in those tissues (HPA tissue IHC). |
| Diffuse staining across tissue or blank areas, with little cell-specific contrast. | Treat this as background until controls support specific staining (general IHC practice). A broad haze does not reproduce the reported cytoplasmic pattern in named placental and endometrial cells (HPA tissue IHC profile). |
| No signal in a placental section despite an otherwise usable stain. | Review whether the section contains the relevant cells and whether controls show that detection worked (general IHC practice). Placental endothelial cells are recorded as high by HPA, but that observation does not guarantee staining in every section or preparation (HPA tissue IHC). |
💡Expected C1QTNF6 appearanceCall an IHC result consistent when cytoplasmic staining identifies placental trophoblasts or endometrial stromal cells, with high placental endothelial and medium endometrial stromal staining reported by HPA; widespread cell-independent color is suspect background (HPA tissue IHC profile and levels; general IHC practice).
| Tissue and cell selection | Placenta provides an HPA-listed high-staining endothelial population; endometrium provides medium-staining stromal cells (HPA tissue IHC). Confirm those cells are present before using either section to judge a negative result (general IHC practice). |
| Secreted protein | C1QTNF6 has a signal peptide at residues 1–46, a mature chain at 47–278, and no transmembrane segment (UniProt Q9BXI9). Interpret location in tissue cautiously: secreted protein and its RNA can appear in different places (HPA tissue IHC reliability description). |
| Antibody and variant context | HPA002042 is reported as IHC Enhanced and ICC Approved (HPA antibody validation). UniProt lists three isoforms (UniProt Q9BXI9). The supplied records do not identify antibody epitope or isoform coverage, so do not infer variant-specific staining. |
| IF/ICC Q: Should nuclear ICC-IF signal define the IHC pattern? | A: No. HPA reports approved nucleoplasmic localization in ICC-IF images from A-431, U-251MG and U2OS, while its tissue IHC profile is cytoplasmic (HPA subcellular ICC-IF; HPA tissue IHC profile). Assess each application against its own evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| Placental endothelial cells show no IHC signal. | The expected cell population may be absent from the sampled area, or the staining run may have failed (HPA tissue IHC; general IHC practice). | Locate endothelial cells morphologically, then check the run’s positive control and antibody/detection steps (general IHC practice). HPA’s high call supports using this population as a comparator, not assuming every section must be positive (HPA tissue IHC). |
| Endometrial stroma looks weaker than placental endothelial cells. | That difference agrees with HPA’s medium stromal and high endothelial ratings (HPA tissue IHC). | Score intensity within the identified cell type and compare sections stained in the same run (general IHC practice). Do not require the endometrial stroma to match the placental endothelial intensity (HPA tissue IHC levels). |
| Adipocytes stain strongly. | The signal conflicts with HPA’s not-detected adipocyte calls and may reflect nonspecific or endogenous detection (HPA tissue IHC; general IHC practice). | Inspect a no-primary control and the distribution of color; review blocking and detection controls if background persists (general IHC practice). Compare adipocytes specifically, rather than assigning a negative call to every cell in adipose tissue or breast (HPA tissue IHC). |
| Predominantly nuclear IHC staining appears. | Nuclear localization is reported for ICC-IF, whereas the tissue IHC profile describes cytoplasmic expression (HPA subcellular ICC-IF; HPA tissue IHC profile). | Record the compartment and cell type, check controls, and avoid scoring nuclear color alone as the expected tissue IHC result (general IHC practice; HPA tissue IHC profile). |
| Diffuse chromogen obscures cell boundaries. | Broad color without cellular contrast can arise from assay background (general IHC practice); it cannot establish the reported cell-specific pattern (HPA tissue IHC profile). | Compare no-primary and positive controls, review blocking and washing, and reassess only where cells remain identifiable (general IHC practice). |
| IF/ICC shows nucleoplasmic signal while IHC looks cytoplasmic. | HPA reports different observations for ICC-IF and tissue IHC (HPA subcellular ICC-IF; HPA tissue IHC profile). | Document application and specimen type with each observation; interpret IHC against tissue IHC evidence and ICC-IF against its separate nucleoplasmic record (HPA tissue IHC; HPA subcellular ICC-IF). |
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: placenta, endothelial cells). Run adipose tissue as the negative comparator, where adipocytes are Not detected; on the placenta slide, use adjacent non-endothelial cells to assess background without assuming they are target-negative (HPA: adipose tissue, adipocytes; HPA: placenta, endothelial cells).
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 a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species, immunoglobulin class and monoclonal or polyclonal format; use knockout tissue or peptide competition, if available, as a biological specificity control (standard IHC practice). For chromogenic placenta IHC, quench endogenous peroxidase and check blood-rich areas for background; block endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected A12414-1 brain IHC caption does not state a fixative (selected-SKU caption). Retrieval dependence is also unreported, so optimize antigen retrieval for the catalog antibody on matched sections (standard IHC practice). The supplied evidence does not establish whether frozen sections or IF are easier than paraffin IHC; placental endogenous peroxidase may complicate chromogenic interpretation (HPA: placenta, endothelial cells; HPA: ICC-IF images in A-431, U-251MG and U2OS; 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 C1QTNF6 staining in paraffin sections by checking retrieval, compartment, cell identity, and controls before comparing signal across samples.
How should I retrieve C1QTNF6 in paraffin sections when staining is weak?
Start with citrate pH 6.0 heat retrieval at 95–98 °C for 20 min (page retrieval setting). Keep section thickness, heating, and cooling consistent across slides so differences in staining remain interpretable (standard IHC practice). If signal is weak, check that sections stayed submerged and compare a modestly longer heating time on adjacent sections while watching for tissue damage (standard IHC practice). The catalog image shows human brain staining at 10 µg/mL, but its caption does not report retrieval conditions (A12414-1 caption). Judge each change against a positive control and a primary antibody omission control (standard IHC practice).
Could fixation explain variable C1QTNF6 staining between paraffin blocks?
C1QTNF6 specific sensitivity to fixation is unknown from the supplied evidence, so record the fixative and processing history for every block (A12414-1 caption; standard IHC practice). The catalog image reports human brain staining at 10 µg/mL but gives no fixative or fixation duration (A12414-1 caption). Compare adjacent sections from blocks with known processing histories using the same citrate pH 6.0 retrieval and detection run (page retrieval setting; standard IHC practice). Include a consistently processed positive control to detect run variation before attributing a difference to fixation (standard IHC practice). Do not infer fixation sensitivity from C1QTNF6 secretion or glycosylation (UniProt Q9BXI9 topology and processing).
Which staining compartment should count as plausible C1QTNF6 signal?
C1QTNF6 has a signal peptide, lacks a transmembrane segment, and is annotated as secreted, so assess intracellular and extracellular staining separately (UniProt Q9BXI9). HPA describes cytoplasmic staining in placental trophoblasts and endometrial stroma, and records high placental endothelial staining (HPA tissue IHC). HPA also reports nucleoplasmic localisation from cell imaging, creating a compartment discrepancy that tissue staining alone cannot resolve (HPA subcellular; HPA tissue IHC). Record the stained cell type and compartment before scoring intensity, and compare them with an adjacent primary antibody omission control (standard IHC practice). Isolated nuclear staining needs independent validation before being assigned to C1QTNF6 (HPA subcellular; UniProt Q9BXI9 secretion).
Could isoforms or processing explain a change in C1QTNF6 staining?
C1QTNF6 has 3 annotated isoforms, a signal peptide spanning residues 1–46, and a mature chain spanning 47–278 (UniProt Q9BXI9). Its collagen like region spans 97–138, its C1q domain spans 139–259, and glycosylation is annotated at residue 91 (UniProt Q9BXI9). The supplied catalog caption does not identify the antibody epitope, so it cannot establish which isoforms or processed forms are detected (A12414-1 caption). Check the catalog antibody’s immunogen or epitope documentation before interpreting a negative section as loss of every isoform (standard IHC practice; UniProt Q9BXI9 isoforms). Compare staining patterns with an independent antibody whose epitope is documented, if available (standard IHC practice).
How can I check C1QTNF6 localisation by multiplex immunofluorescence?
For a placental comparison, pair C1QTNF6 with an endothelial marker such as CD31 because HPA records high staining in placental endothelial cells (HPA tissue IHC; standard IF practice). Use spectrally separated fluorophores and consider a far red channel for C1QTNF6 when tissue autofluorescence is prominent in shorter wavelength channels (standard IF practice). Because C1QTNF6 lacks a transmembrane segment, apply a mild detergent permeabilisation step when testing intracellular signal, and omit detergent on a parallel section when examining accessible extracellular deposits (UniProt Q9BXI9 topology; standard IF practice). HPA cell imaging reports nucleoplasmic localisation, so capture nuclear counterstain and compartment resolved images (HPA subcellular; standard IF practice). Include single stain and primary antibody omission controls for channel bleed through and background (standard IF practice).
What should I check when C1QTNF6 staining appears diffuse or widespread?
First compare the stained slide with a primary antibody omission control and inspect unstained tissue regions for pigment or precipitate (standard IHC practice). Block endogenous peroxidase before chromogenic detection, and control development time so DAB does not obscure cellular boundaries (standard IHC practice). The catalog caption reports 10 µg/mL antibody in human brain, but provides no evidence that this concentration is optimal for other tissues (A12414-1 caption). Titrate the catalog antibody around a documented working concentration using identical retrieval and detection conditions across adjacent sections (standard IHC practice). Because C1QTNF6 is secreted, evaluate diffuse extracellular signal separately from cell associated signal rather than scoring both as positive cells (UniProt Q9BXI9; standard IHC practice).
How should I quantify C1QTNF6 across tissues with different cell mixtures? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring because C1QTNF6 is secreted and HPA reports staining in distinct placental and endometrial populations (UniProt Q9BXI9; HPA tissue IHC). For cell associated chromogenic signal, record the percentage of positive cells and an H score using intensity categories 0–3 (standard IHC scoring practice). If extracellular deposits are the endpoint, measure positive area or signal density per mm² of viable tissue instead of assigning deposits to nearby cells (standard image analysis practice). Normalise cell counts to the number of eligible cells and area measurements to annotated viable tissue area (standard image analysis practice). Keep retrieval, imaging, thresholds, and DAB development consistent across the comparison (standard IHC practice).
How can I distinguish true C1QTNF6 staining from artefact?
Treat a positive result as more convincing when the expected cell population stains reproducibly and primary antibody omission controls remain clear (HPA tissue IHC; standard IHC practice). HPA reports high placental endothelial staining and medium staining in endometrial stromal cells, offering defined populations for comparison (HPA tissue IHC). Scrutinise isolated nuclear signal because HPA cell imaging reports nucleoplasmic localisation while tissue IHC describes cytoplasmic expression (HPA subcellular; HPA tissue IHC). Exclude section edges, necrotic regions, pigment, and residual endogenous peroxidase signal before assigning positive cells (standard IHC practice). Interpret extracellular staining cautiously because secreted C1QTNF6 may be observed away from its producing cells (UniProt Q9BXI9; HPA reliability description).
Boster reagents
Best C1QTNF6 / Complement C1q tumor necrosis factor-related protein 6 IHC Antibodies
The catalog includes an IHC-P antibody with a human brain tissue image and an IF/ICC antibody with HeLa cell images; listed reactivity spans human, mouse and rat (catalog applications, reactivity and image captions).
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, with an IHC image of human brain tissue at 10 μg/mL (catalog applications; A12414-1 image caption). A12414 lists IF and ICC, with a HeLa immunocytochemistry image at 10 μg/mL (catalog applications; A12414 image caption).
Which to pick: Choose A12414-1 for paraffin-section IHC: it lists IHC-P and human, mouse and rat reactivity, and its IHC image shows human brain tissue; the fixative is unreported (catalog applications and reactivity; A12414-1 image caption). Choose A12414 for IF/ICC when human or mouse reactivity fits, using its HeLa cell images as the illustrated evidence (catalog applications and reactivity; A12414 image captions). For rat reactivity, choose A12414-1; clonality is unreported for both SKUs (catalog reactivity 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 staining in tissue (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: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Secretion can uncouple staining from local expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; check epitope against mature chain 47–278 (UniProt); Regulation: Enhanced in endometrium and placenta (HPA tissue RNA). Evidence: UniProt Q9BXI9, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm., Datasheet A12414-1, PMC7585664, PMC8670214. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/C1QTNF6/immunohistochemistry-ihc-guide.