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TFCP2L1 / Transcription factor CP2-like protein 1 · IHC design guide
Design Immunohistochemistry for TFCP2L1
Plan chromogenic TFCP2L1 IHC around the reported nucleolar tissue pattern (HPA tissue IHC). Use duodenal glandular cells as a positive reference and adipocytes as a negative reference, while interpreting staining cautiously because it shows low consistency with RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TFCP2L1 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 | Nucleolar staining reported in several tissues, including kidney (HPA tissue IHC) | |
| Staining pattern | Nucleolar signal in kidney tubule cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Duodenum+4 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 A08278) | |
| Caveat | Staining and RNA expression show low consistency (HPA tissue IHC) | |
| Regulation | Kidney and salivary RNA is tissue enhanced (HPA tissue RNA) | |
| Isoform / epitope | No isoforms or processing reported; one 1–479 chain (UniProt) |
Section 1
Recommended TFCP2L1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with the published breast tissue FFPE protocol (PMC13562584).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A08278); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-TFCP2L1, 2.5 μg/mL (datasheet A08278) |
| 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 | TFCP2L1-positive staining in glandular cells of duodenum (HPA tissue IHC: High). HPA tissue profile: Nucleolar expression in several tissues, including kidney. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min (page antigen retrieval; UniProt Q9NZI6: nucleus).
Section 2
What Is the Expected TFCP2L1 Staining Pattern?
TFCP2L1 is a nuclear protein with no transmembrane segment (UniProt Q9NZI6). In paraffin-section IHC, expect staining mainly within nuclei of selected epithelial and glandular cells, including duodenal glandular cells and esophageal squamous epithelial cells (HPA tissue IHC). HPA describes a nucleolar pattern in several tissues, including kidney, but rates tissue IHC reliability uncertain because antibody staining has low consistency with RNA expression (HPA tissue IHC).
| Discrete nuclear or nucleolar staining in duodenal glandular cells or esophageal squamous epithelial cells. | Consistent with HPA's high staining observations in those cells and its nucleolar tissue profile (HPA tissue IHC). Assess the pattern in the named cells; HPA rates overall tissue IHC reliability uncertain (HPA tissue IHC). |
| Strong, widespread membrane staining or predominantly diffuse cytoplasmic staining. | Treat as a suspect compartment pattern: UniProt places TFCP2L1 in the nucleus and reports no transmembrane segment (UniProt Q9NZI6). Check antibody specificity and detection controls before scoring it as positive (standard IHC practice). |
| Staining concentrated in cells outside the expected cell population, such as adipocytes in adipose tissue. | HPA reports adipocytes as not detected (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity; compare a no-primary control and the expected positive cell population (standard IHC practice). |
| Uniform haze or precipitate across tissue and empty areas of the section. | This distribution is difficult to assign to TFCP2L1-positive cells. Review blocking, washes, chromogen development and no-primary controls for nonspecific background (standard IHC practice); HPA's tissue profile is cell-selective (HPA tissue IHC). |
| No staining in duodenal glandular cells despite an intact, readable section. | HPA reports high staining there, so check the staining run, antigen retrieval and antibody conditions (HPA tissue IHC; standard IHC practice). A failed result does not establish target absence, especially given HPA's uncertain IHC reliability (HPA tissue IHC). |
💡Expected TFCP2L1 appearanceCall a result positive when discrete nuclear, often nucleolar, signal occurs in the relevant cells—for example, high staining in duodenal glandular cells; widespread membrane signal or uniform haze is suspect (HPA tissue IHC; UniProt Q9NZI6; standard IHC practice).
| Compartment and image resolution | UniProt assigns TFCP2L1 to the nucleus; HPA describes nucleolar tissue staining (UniProt Q9NZI6; HPA tissue IHC). HPA ICC-IF refines its approved main location to the nucleoli fibrillar center, a detail that routine chromogenic IHC may not resolve (HPA subcellular; standard IHC practice). |
| Cell and tissue selection | HPA records high staining in duodenal glandular and esophageal or vaginal squamous epithelial cells, and medium staining in kidney tubule cells and salivary glandular cells (HPA tissue IHC). Compare named cell populations, not whole-section color (standard IHC practice). |
| Strength of tissue evidence | HPA labels tissue IHC uncertain because staining and RNA expression have low consistency; its RNA specificity is tissue enhanced in kidney and salivary gland (HPA tissue IHC). Treat staining levels as observed patterns rather than independently confirmed expression in every specimen. |
| Antibody validation by application | HPA029708 is rated uncertain for IHC and approved for ICC (HPA antibodies). HPA ICC-IF reports mitochondrial localization as uncertain (HPA subcellular). Neither ICC rating nor that additional location validates mitochondrial staining in paraffin-section IHC. |
| Protein topology and processing | UniProt lists one chain spanning residues 1–479, no signal peptide or propeptide, no transmembrane segment, and no annotated glycosylation sites (UniProt Q9NZI6). Those entries provide no basis to expect a surface or secreted IHC pattern. |
| Retrieval and detection controls | Antigen retrieval and detection conditions can affect paraffin-section staining in general (standard IHC practice). The supplied UniProt and HPA records give no TFCP2L1-specific fixation sensitivity or retrieval setting; assess each run against tissue and no-primary controls. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells show little or no nuclear signal. | The run may have inadequate retrieval or detection, or the tissue result may differ from HPA's uncertain IHC profile (standard IHC practice; HPA tissue IHC). | Check section integrity and run controls, then review retrieval and antibody conditions using duodenal glandular cells as an observed high reference (standard IHC practice; HPA tissue IHC). |
| Membrane staining dominates the section. | This conflicts with nuclear localization and the absence of a transmembrane segment (UniProt Q9NZI6); nonspecific antibody or detection staining is possible (standard IHC practice). | Inspect a no-primary control and the nuclear pattern in the expected cells before assigning a positive score (standard IHC practice; HPA tissue IHC). |
| Adipocytes stain as strongly as the proposed positive cells. | HPA reports adipocytes in adipose tissue as not detected; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; standard IHC practice). | Compare no-primary staining, review blocking and chromogen conditions, and score the named cell populations separately (standard IHC practice). |
| The entire slide has diffuse brown background. | Nonspecific binding, endogenous detection activity or excessive chromogen development can obscure cellular localization (standard IHC practice). | Check the no-primary control; review blocking, wash steps and development time, then reassess nuclear signal above background (standard IHC practice; UniProt Q9NZI6). |
| Kidney tubule cells stain, but the pattern is hard to classify. | HPA reports medium kidney tubule staining and a nucleolar tissue profile, while overall tissue IHC reliability remains uncertain (HPA tissue IHC). | Score tubule cells and subcellular distribution separately; compare run controls and avoid treating an unresolved nucleolar detail as a failed chromogenic stain (standard IHC practice; HPA tissue IHC). |
| An ICC-IF image suggests mitochondrial signal that the IHC slide lacks. | HPA calls mitochondria an uncertain additional ICC-IF location; its approved main ICC-IF location is the nucleoli fibrillar center (HPA subcellular). | Interpret paraffin-section IHC using its nuclear or nucleolar tissue pattern and IHC controls; do not require mitochondrial signal for a positive call (HPA tissue IHC; HPA subcellular; standard IHC practice). |
Experiment evidence
IHC & IF images showing TFCP2L1 staining
A08278 · IHCImmunohistochemistry of TFCP2L1 in rat colon tissue with TFCP2L1 antibody at 2.5 μg/mL.
A08278 · IFImmunofluorescence of TFCP2L1 in rat colon tissue with TFCP2L1 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000115112-TFCP2L1/tissue/cerebral+cortex#imid_7637016
HPA · Salivary glandHuman Protein Atlas IHC, Salivary gland. Source: https://www.proteinatlas.org/ENSG00000115112-TFCP2L1/tissue/salivary+gland#imid_7636980
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000115112-TFCP2L1/tissue/colon#imid_7636950
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000115112-TFCP2L1/tissue/liver#imid_7636943
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA029708; green = TFCP2L1, blue = nucleus, red = microtubules.
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA029708; green = TFCP2L1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TFCP2L1 IHC & IF
🧪Run duodenum first; its glandular cells should stain (HPA: High in duodenal glandular cells). Use adipose tissue as a negative tissue control (HPA: Not detected in adipocytes); on the duodenum slide, treat non-glandular cells as internal negatives only if they show background-level staining, since their TFCP2L1 status is not specified by the supplied HPA row (HPA: duodenal glandular-cell entry).
Positive control tissue: Duodenum (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TFCP2L1 in RT-4, U-251MG, U2OS, with annotated localisation: Nucleoli fibrillar center (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species, clonality and concentration; confirm target specificity with TFCP2L1-knockout material or an immunizing-peptide block if available (standard IHC practice). For duodenal chromogenic IHC, quench endogenous peroxidase and check for endogenous biotin background 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 A08278 rat-colon IHC caption does not state a fixative (caption: fixative not stated). Antigen-retrieval dependence is unreported; optimize retrieval on paraffin sections using the positive and negative tissue controls (standard IHC practice; HPA: High in duodenal glandular cells; HPA: Not detected in adipocytes). The supplied evidence does not establish whether frozen sections or IF are easier; duodenal endogenous enzyme or biotin activity may complicate chromogenic scoring (standard IHC practice).
HPA tissue IHC evidence for TFCP2L1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Vagina | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | Medium | 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 → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TFCP2L1 IHC Tips
Use nuclear staining, tissue context, and matched controls to troubleshoot TFCP2L1 chromogenic IHC (UniProt Q9NZI6: nucleus; HPA tissue IHC: uncertain reliability).
How should I retrieve TFCP2L1 in paraffin sections when nuclear staining is weak?
Start with Tris-EDTA pH 9.0 heat-induced epitope retrieval at 95–98 °C for 20 min (page retrieval setting). Let sections cool in buffer for about 20 min, then compare stained and primary-omission slides under the same detection conditions (standard IHC practice). If nuclear staining remains weak, test a shorter or longer heating interval on adjacent sections while holding antibody concentration and detection time constant; a different buffer should be a later comparison, not the starting condition (standard IHC practice). Assess nuclear signal in the relevant cell population because TFCP2L1 is annotated as nuclear (UniProt Q9NZI6: nucleus).
Could fixation explain weak or uneven TFCP2L1 staining?
Target-specific fixation sensitivity is unknown because the supplied TFCP2L1 tissue image does not state a fixative (A08278 tissue-IHC caption: fixative not stated). Record fixative, time before fixation, and fixation duration for each specimen, then compare adjacent sections processed together (standard IHC practice). If staining differs across blocks, first check section adhesion, retrieval consistency, and detection timing before attributing the difference to TFCP2L1 biology (standard IHC practice). Keep the rat-colon image’s 2.5 µg/mL antibody concentration as an image-specific reference rather than evidence for any fixation condition (A08278 tissue-IHC caption: rat colon, 2.5 µg/mL).
Where should convincing TFCP2L1 staining appear in tissue sections?
Prioritize a nuclear pattern when evaluating chromogenic TFCP2L1 staining (UniProt Q9NZI6: nucleus). Tissue IHC also reports nucleolar staining in several tissues, including kidney, but assigns the overall staining profile uncertain reliability because it correlates poorly with RNA expression (HPA tissue IHC: nucleolar profile; uncertain reliability). Score nuclear and nucleolar signal separately from diffuse cytoplasmic or luminal color, and inspect cell borders at high magnification before calling a cell positive (standard IHC practice). Use matched morphology and a primary-omission control to investigate staining outside nuclei; the protein has no annotated transmembrane segment (UniProt Q9NZI6: topology).
How can epitope uncertainty affect TFCP2L1 IHC interpretation?
The supplied record lists a 479-aa chain, zero isoforms, and a Grh/CP2 DNA-binding domain at residues 43–280 (UniProt Q9NZI6: processing, isoforms, domain). Those annotations do not locate the catalog antibody’s epitope, so request its immunogen or epitope information before making domain-specific claims (supplied antibody evidence: epitope not stated). If two validated antibodies recognize disclosed, distinct epitopes, concordant staining in adjacent sections can strengthen interpretation (standard IHC practice). Compare their nuclear patterns under matched retrieval and detection conditions, while treating disagreement as a prompt to investigate accessibility or specificity rather than evidence of an unlisted isoform (standard IHC practice).
How should I plan a TFCP2L1 multiplex IF check?
Use IF as an independent localisation check, pairing TFCP2L1 with a marker for the cell population being evaluated and a nuclear counterstain (standard IF practice). Choose separated fluorophores and place the weaker signal in a channel with low tissue autofluorescence; collect single-stain and unstained controls to assess bleed-through and background (standard IF practice). Because TFCP2L1 is nuclear and has no transmembrane segment, use controlled permeabilisation to allow access to the intracellular epitope, then compare it with a milder condition if morphology suffers (UniProt Q9NZI6: nucleus, topology; standard IF practice). HPA reports a nucleolar fibrillar-center location in ICC/IF, while its additional mitochondrial location is uncertain (HPA subcellular: approved main location; uncertain additional location).
What should I check when TFCP2L1 chromogenic staining is diffuse?
Run a primary-omission control and inspect whether color persists in the same structures; persistence points to detection-system or tissue background (standard IHC practice). For peroxidase detection, verify the peroxide block and compare a shorter DAB development time, using identical timing across test sections (standard IHC practice). Recheck blocking, wash stringency, and antibody concentration; 2.5 µg/mL is reported for the selected rat-colon image, not established as a universal working concentration (A08278 tissue-IHC caption). Resolve diffuse color against the expected nuclear compartment and the reported, albeit uncertain, nucleolar tissue pattern before scoring it as TFCP2L1 (UniProt Q9NZI6: nucleus; HPA tissue IHC: nucleolar profile, uncertain reliability).
How should I quantify TFCP2L1 across sections with different staining intensities? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and nuclear compartment before analysis, then report the percentage of positive nuclei and an intensity-based H-score on a consistent 0–300 scale (standard IHC practice). If spatial abundance matters, add positive nuclei per mm² of viable tissue, normalized to the sampled tissue area or total nuclei in the same compartment (standard IHC practice). Apply one positivity threshold, exposure-independent image processing, and matched DAB development across the comparison set (standard IHC practice). Report nucleolar signal separately when present, and avoid treating the HPA tissue profile as a calibration standard because its reliability is uncertain (HPA tissue IHC: nucleolar profile, uncertain reliability).
How do I distinguish true TFCP2L1 signal from tissue artefact?
Favor reproducible nuclear staining in morphologically intact cells, with nucleolar staining recorded separately (UniProt Q9NZI6: nucleus; HPA tissue IHC: nucleolar profile). HPA reports medium staining in colon glandular cells and kidney tubular cells, but rates its overall tissue-IHC reliability uncertain, so these patterns are context rather than proof for an individual section (HPA tissue IHC: colon and kidney levels; uncertain reliability). Investigate signal confined to section edges, necrotic areas, lumina, or primary-omission controls before interpreting it as expression (standard IHC practice). Check the peroxidase block when color follows endogenous enzyme activity, and require agreement between cellular location, morphology, and controls before scoring (standard IHC practice).
Boster reagents
Best TFCP2L1 / Transcription factor CP2-like protein 1 IHC Antibodies
A08278 has rat-colon IHC and IF images (A08278 image captions) and lists human, mouse, and rat reactivity (catalog: A08278 reactivity).
Real IHC data
Anti-TFCP2L1 Antibody
Cat # A08278
A08278 lists IHC-P and shows rat-colon IHC at 2.5 μg/mL (catalog: A08278 applications; A08278 IHC caption). It also lists IF and shows rat-colon IF at 20 μg/mL (catalog: A08278 applications; A08278 IF caption).
Which to pick: For paraffin-section tissue IHC, choose A08278: it lists IHC-P and has its own rat-colon IHC image at 2.5 μg/mL; the fixative is unreported (catalog: A08278 applications; A08278 IHC caption). For IF, A08278 has a rat-colon image at 20 μg/mL; ICC-specific validation is unreported (A08278 IF caption; catalog: A08278 applications). For cross-species work, A08278 lists human, mouse, and rat reactivity, though its shown IHC and IF images are from rat colon; clonality is unreported (catalog: A08278 reactivity and clone; A08278 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NZI6 (TF2L1_HUMAN, Transcription factor CP2-like protein 1).
- Human Protein Atlas. TFCP2L1 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. TFCP2L1 subcellular location (ICC-IF): Mainly localized to the nucleoli fibrillar center. In addition localized to the mitochondria..
- Human Protein Atlas. TFCP2L1 antibody validation summary (1 antibodies).
- TFCP2L1 drives stemness and enhances their resistance to Sorafenib treatment by modulating the NANOG/STAT3 pathway in hepatocellular carcinoma. Oncogenesis 2024 — PMC11392926.
- Phosphorylation of TFCP2L1 by CDK1 is required for stem cell pluripotency and bladder carcinogenesis. EMBO molecular medicine 2020 — PMC6949511.
- TFCP2L1 suppresses breast cancer progression by promoting ferroptosis through direct and PI3K/AKT-mediated inhibition of GPX4. Cell death discovery 2026 — PMC13562584.
- Transcription factor TFCP2L1 patterns cells in the mouse kidney collecting ducts. eLife 2017 — PMC5484618.
- PubMed PMID:10644752 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TFCP2L1 (Transcription factor CP2-like protein 1). Expected localisation Nucleolar staining reported in several tissues, including kidney (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A08278 at 2.5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Duodenum (Glandular 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 A08278); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-TFCP2L1, 2.5 μg/mL (datasheet A08278); 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 A08278); Caveat: Staining and RNA expression show low consistency (HPA tissue IHC); Isoform / epitope: No isoforms or processing reported; one 1–479 chain (UniProt); Regulation: Kidney and salivary RNA is tissue enhanced (HPA tissue RNA). Evidence: UniProt Q9NZI6, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the nucleoli fibrillar center. In addition…, Datasheet A08278, PMC13562584. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TFCP2L1/immunohistochemistry-ihc-guide.