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DPF3 / Zinc finger protein DPF3 · IHC design guide
Design Immunohistochemistry for DPF3
Plan chromogenic DPF3 IHC around the nuclear staining reported in neurons, ovary, testis and muscle (HPA tissue IHC). This guide covers tissue controls, consistent fixation and interpretation of the IHC-validated antibody’s staining, with HPA’s medium RNA concordance in mind (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DPF3 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 | Nuclear staining in positive tissues (HPA tissue IHC) | |
| Staining pattern | Nuclei of neurons, follicle cells, spermatocytes and myocytes (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining has medium concordance with RNA data (HPA tissue IHC) | |
| Regulation | Staining varies by tissue and cell type (HPA tissue IHC) | |
| Isoform / epitope | Five isoforms; epitope coverage needs confirmation (UniProt) |
Section 1
Recommended DPF3 IHC & IF Protocols
The catalog antibody protocol is paired with one published DPF3 IHC protocol reporting primary dilution and antigen retrieval without retrieval conditions (PMC8897700 methods).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A09485); 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-DPF3, 5 μg/mL (datasheet A09485) |
| 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 | DPF3-positive staining in gLUC cells - nucleus of cerebellum (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in neurons, ovary, testis and heart and skeletal myocytes. 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 rule: nuclear antigen).
Section 2
What Is the Expected DPF3 Staining Pattern?
DPF3 is a nuclear chromatin regulator with no transmembrane segment (UniProt Q92784: subcellular location, topology). In paraffin-section IHC, expect staining chiefly in neuronal nuclei and in the nuclei of heart and skeletal myocytes; HPA also describes nuclear expression in ovary and testis (HPA: tissue profile). HPA rates the tissue IHC evidence Enhanced, with medium staining–RNA consistency and external verification pending (HPA: tissue reliability).
| High nuclear staining in cerebellar GLUC cells; neuronal staining in cerebral cortex (HPA: tissue IHC). | This fits the strongest supplied neural examples. Assess signal in the annotated cells and compartment: HPA explicitly calls cerebellar GLUC nuclei high, while its cortex entry calls neuronal cells high (HPA: tissue IHC). |
| Nuclear staining in pachytene spermatocytes, cardiomyocytes or ovarian follicle cells (HPA: tissue IHC). | These are supported cell populations, with high staining in pachytene spermatocytes and medium staining in cardiomyocytes and follicle cells (HPA: tissue IHC). Compare like cells; a whole-tissue average can hide their distribution (standard IHC interpretation). |
| Predominantly cytoplasmic, membrane or extracellular signal, without convincing nuclear staining. | Treat this as discordant with DPF3's nuclear annotation and HPA's nuclear tissue profile (UniProt Q92784: subcellular location; HPA: tissue profile). Recheck localisation against a nuclear counterstain before calling it DPF3 (standard IHC practice). |
| Staining in an HPA-undetected cell population, especially if it follows vessels, pigment or section edges. | For example, HPA reports adipocytes in adipose tissue and respiratory epithelial cells in bronchus as not detected (HPA: tissue IHC). Such signal warrants checks for cross-reactivity, endogenous chromogenic activity or deposition artefact (standard IHC practice). |
| Diffuse colour across cells and stroma, or no nuclear signal in cerebellar GLUC cells. | Diffuse colour obscures compartment scoring and suggests background (standard IHC interpretation). Absent signal in a supplied high nuclear reference raises a run or detection concern, though one negative slide cannot establish absent DPF3 expression (HPA: cerebellum tissue IHC; standard IHC practice). |
💡Expected DPF3 appearanceCall a positive result when staining is predominantly nuclear in the supported cells, especially high in cerebellar GLUC cells or pachytene spermatocytes; widespread non-nuclear colour is suspect (HPA: tissue IHC and profile; UniProt Q92784: subcellular location; standard IHC interpretation).
| Reference-cell selection | Use the annotated cell population, not an entire organ, to judge concordance. HPA reports high nuclear staining in cerebellar GLUC cells, but only low staining in hippocampal neuronal cells and skeletal myocytes (HPA: tissue IHC). |
| Tissue evidence and antibody validation | HPA's tissue pattern has Enhanced reliability but only medium staining–RNA consistency, with external verification pending (HPA: tissue reliability). HPA066790 has Enhanced IHC validation; HPA070601 has no listed IHC status, so its Enhanced ICC status should not be read as IHC validation (HPA: antibody validation). |
| Cellular location and processing | A predominantly nuclear result agrees with the UniProt nucleus annotation and HPA tissue profile (UniProt Q92784: subcellular location; HPA: tissue profile). UniProt lists no transmembrane segment, signal peptide or propeptide, and a single 1–378 chain (UniProt Q92784: topology and processing). |
| Isoforms and modified residue | UniProt lists 5 isoforms and phosphoserine at residue 323 (UniProt Q92784: isoforms, modified residues). The supplied evidence does not map an antibody epitope to an isoform or this residue; do not infer staining differences from them (supplied UniProt and HPA records). |
| Antigen retrieval and fixation | Retrieval conditions can be optimised as a general paraffin-IHC workflow step (standard IHC practice). No supplied target-specific evidence establishes that fixation or retrieval changes DPF3 staining, so report the tested condition without attributing a staining difference to DPF3-specific sensitivity (supplied UniProt and HPA records). |
| IF/ICC Q: where should fluorescence appear? | A: HPA reports DPF3 in the nucleoplasm, with an Enhanced main-location assessment and ICC-IF images from BJ, PC-3 and U2OS cells (HPA: subcellular). This is an IF/ICC localisation reference, not a paraffin-IHC protocol (HPA: subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| No nuclear staining in a cerebellar reference section. | The reference is discordant with HPA's high nuclear GLUC-cell result; failed staining or detection is possible (HPA: cerebellum tissue IHC; standard IHC practice). | Confirm the annotated cells are present; review the run control, antibody dilution, retrieval and detection steps before interpreting the specimen as negative (standard IHC practice). |
| Only cytoplasmic or membrane colour is visible. | This conflicts with the nuclear location reported by UniProt and HPA; nonspecific binding or misplaced chromogen may explain the colour (UniProt Q92784: subcellular location; HPA: tissue profile; standard IHC interpretation). | Compare with the nuclear counterstain and a reagent control, then score nuclear and non-nuclear signal separately (standard IHC practice). |
| Broad staining appears in cells expected to be undetected. | HPA lists adipocytes in adipose tissue and respiratory epithelial cells in bronchus as not detected; cross-reactivity or endogenous detection activity is possible (HPA: tissue IHC; standard IHC practice). | Check a reagent control, endogenous-enzyme blocking and chromogen deposition; reassess the exact cell type rather than the organ label (standard chromogenic-IHC practice). |
| Diffuse colour makes nuclei hard to distinguish. | Nonspecific background or excessive detection can obscure compartment assignment (standard IHC interpretation). | Review blocking, antibody dilution, washes and detection development; use a nuclear counterstain to judge whether a discrete nuclear component remains (standard IHC practice). |
| Heart or skeletal muscle looks weaker than cerebellum. | HPA reports medium cardiomyocyte staining and low skeletal-myocyte staining, versus high cerebellar GLUC-cell staining (HPA: tissue IHC). | Score the relevant cells against their own HPA level; do not require muscle to match the cerebellar reference intensity (HPA: tissue IHC; standard IHC interpretation). |
| An ICC-validated antibody is being considered for paraffin IHC. | HPA070601 has Enhanced ICC validation but no listed IHC status; the validation entries cover different applications (HPA: antibody validation). | Use an IHC-validated antibody as the paraffin-section reference; HPA066790 has Enhanced IHC validation, while an ICC result alone does not establish IHC performance (HPA: antibody validation). |
Experiment evidence
IHC & IF images showing DPF3 staining
A09485 · IHCImmunohistochemistry of DPF3 in human brain tissue with DPF3 antibody at 5 μg/mL.
A09485 · IFImmunofluorescence of DPF3 in human brain tissue with DPF3 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000205683-DPF3/tissue/cerebral+cortex#imid_18985821
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000205683-DPF3/tissue/colon#imid_18985739
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000205683-DPF3/tissue/liver#imid_18985740
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000205683-DPF3/tissue/kidney#imid_18985749
![HPA ICC-IF BJ [Human fibroblast] — DPF3 (HPA066790)](https://images.proteinatlas.org/66790/1544_B12_2_blue_red_green.jpg)
HPA · ICC-IF BJ [Human fibroblast]Human Protein Atlas immunofluorescence, BJ [Human fibroblast], antibody HPA066790; green = DPF3, blue = nucleus, red = microtubules.
![HPA ICC-IF BJ [Human fibroblast] — DPF3 (HPA066790)](https://images.proteinatlas.org/66790/1544_B12_3_blue_red_green.jpg)
HPA · ICC-IF BJ [Human fibroblast]Human Protein Atlas immunofluorescence, BJ [Human fibroblast], antibody HPA066790; green = DPF3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DPF3 IHC & IF
🧪Run cerebellum first and look for nuclear staining in GLUC cells (HPA: High in cerebellar GLUC cell nuclei). Use adipose tissue as the negative tissue; its adipocytes should lack staining (HPA: Not detected in adipocytes). On the positive slide, cells without nuclear signal provide an internal background reference; do not assign them a specific cell identity from the supplied data (standard IHC practice).
Positive control tissue: Cerebellum (GLUC cells - nucleus, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show DPF3 in BJ [Human fibroblast], PC-3, U2OS, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a matched isotype control chosen for the primary antibody’s host species and clonality, and DPF3 knockout material or a validated peptide-block control (standard IHC practice). Quench endogenous peroxidase for chromogenic detection and distinguish brain pigment from specific nuclear staining; for IF, check tissue autofluorescence with an unstained control (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window and antigen-retrieval dependency are unreported in the supplied evidence; optimize retrieval empirically for paraffin sections (standard IHC practice). The selected A09485 human-brain IHC caption gives 5 µg/mL but does not report a fixative (A09485 caption: 5 µg/mL; fixative unreported). The evidence does not establish that frozen sections or IF are easier for DPF3; brain pigment can complicate interpretation of chromogenic staining (standard IHC practice).
HPA tissue IHC evidence for DPF3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Cerebellum | GLUC cells - nucleus | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Kidney | Proximal tubules (cell body) | High | Protein (IHC) | HPA → |
| Testis | Pachytene spermatocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | 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 → |
| Colon | Endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced DPF3 IHC Tips
Troubleshoot DPF3 chromogenic IHC by checking nuclear staining, tissue context and matched controls before interpreting signal intensity.
How should I retrieve DPF3 in paraffin sections when nuclear staining is weak?
Use Tris-EDTA pH 9.0 heat-induced epitope retrieval at 95–98 °C for 20 min (page retrieval rule: nuclear antigen). Cool sections gradually in retrieval buffer, then rinse consistently before blocking (standard IHC practice). Assess nuclear staining in a positive tissue section alongside a no-primary control; DPF3 is a nuclear protein (UniProt Q92784: nucleus). If staining remains weak, test citrate pH 6.0 as a fallback on adjacent sections, holding detection and exposure conditions constant (standard IHC optimization practice). Record any improvement against background and tissue preservation, since stronger chromogen alone does not establish target-specific staining (standard IHC interpretation practice).
Could fixation explain weak or uneven DPF3 staining?
Target-specific DPF3 sensitivity to fixative type or duration is unknown from the supplied evidence (evidence payload: no target-specific fixation data). The selected antibody image shows human brain staining at 5 µg/mL, but its caption does not state the fixative (A09485 tissue-IHC caption). Compare sections with documented, matched fixation and processing before changing antibody concentration (standard IHC practice). If fixation varies, run a small retrieval comparison on serial sections and judge nuclear signal together with morphology and background (standard IHC optimization practice). Do not attribute a difference to DPF3 biology until the processing difference and assay controls have been assessed (standard IHC interpretation practice).
What staining pattern should count as plausible DPF3 localisation?
Prioritise nuclear staining: DPF3 is annotated in the nucleus, and subcellular imaging places it in the nucleoplasm (UniProt Q92784: nucleus; HPA subcellular: nucleoplasm). Nuclear staining in neurons and heart myocytes is consistent with the reported tissue profile (HPA tissue IHC: neurons and heart myocytes). Score nuclei separately from cytoplasmic or extracellular chromogen, using a counterstain light enough to preserve that distinction (standard IHC scoring practice). DPF3 has no annotated transmembrane segment, so a sharp membrane-only pattern needs independent validation (UniProt Q92784: topology). Compare the same cell compartment across serial sections because section thickness and chromogen development can alter apparent signal intensity (standard IHC practice).
Can a negative IHC result exclude every DPF3 isoform?
No: the record lists 5 DPF3 isoforms, while the supplied antibody evidence does not map its epitope across them (UniProt Q92784: isoforms 1–5; A09485 tissue-IHC caption). Check whether the antibody documentation identifies an immunogen or epitope before treating absence of staining as absence of all isoforms (standard antibody validation practice). DPF3 also has a reported phosphoserine at residue 323, but the payload does not establish whether that modification changes antibody binding (UniProt Q92784: modified residue). If the epitope remains unknown, compare an independently validated antibody on serial sections and report the result as antibody-dependent staining (standard IHC validation practice).
How should I adapt DPF3 localisation checks to multiplex IF?
Use a marker for the expected cell type, such as a validated neuronal marker in brain, and inspect whether DPF3 signal occupies its nuclei (HPA tissue IHC: neuronal nuclear expression; standard IF multiplexing practice). Choose spectrally separated fluorophores and place the weaker DPF3 signal in a red or far-red channel after checking tissue autofluorescence and single-stain controls (standard IF practice). Permeabilise sufficiently for antibody access to the nuclear epitope; DPF3 has no annotated transmembrane segment (UniProt Q92784: nucleus and topology; standard IF practice). Optimise IF fixation, permeabilisation and antibody concentration independently: the selected brain caption reports IHC staining at 5 µg/mL, without establishing IF conditions (A09485 tissue-IHC caption).
How can I reduce diffuse or misleading DPF3 chromogen?
Compare a no-primary section with the test section to identify detection-system or tissue-derived background (standard IHC control practice). Apply a peroxidase block before HRP/DAB detection, then check whether residual pigment persists in the no-primary control (standard chromogenic IHC practice). Titrate the primary antibody and detection development on matched sections while preserving interpretable nuclear staining (standard IHC optimization practice). The selected tissue image used 5 µg/mL in human brain, but its caption does not establish an optimal concentration for other sections (A09485 tissue-IHC caption). Diffuse cytoplasmic colour should be assessed separately from expected nucleoplasmic signal, especially at tissue edges (HPA subcellular: nucleoplasm; standard IHC interpretation practice).
What is a defensible way to quantify DPF3 IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and nuclear compartment before scoring, since DPF3 is nuclear and reported expression differs across cell types (UniProt Q92784: nucleus; HPA tissue IHC: cell-specific profile). Report the percentage of positive nuclei and, when intensity is reproducible, an H-score from 0–300 using fixed intensity thresholds (standard IHC scoring practice). For spatial comparisons, positive nuclei per mm² can supplement the percentage, provided the sampled area is recorded (standard digital pathology practice). Normalise positive counts to all evaluable nuclei of the same cell type and exclude folds, necrosis and cut edges using prespecified rules (standard IHC quantification practice).
How do I distinguish convincing DPF3 staining from artefact?
A convincing result shows reproducible nuclear signal in an appropriate cell population, such as neuronal cells, with acceptable no-primary background (UniProt Q92784: nucleus; HPA tissue IHC: neuronal expression; standard IHC control practice). A membrane-only or extracellular pattern conflicts with the annotated nuclear localisation and lack of a transmembrane segment (UniProt Q92784: localisation and topology). Treat staining confined to cut edges, damaged areas or necrosis as suspect and compare it with intact tissue nearby (standard IHC interpretation practice). Check a no-primary section for endogenous enzyme activity before attributing DAB colour to DPF3 (standard chromogenic IHC practice). HPA reports enhanced reliability but pending external verification, so corroborate unexpected patterns independently (HPA tissue IHC: reliability).
Boster reagents
Best DPF3 / Zinc finger protein DPF3 IHC Antibodies
A09485 has human brain IHC and IF images (catalog image captions) and lists human and mouse reactivity (catalog reactivity).
Real IHC data
Anti-Zinc finger protein DPF3 DPF3 Antibody
Cat # A09485
A09485 is listed for IHC-P and has a human brain IHC image at 5 μg/mL (catalog applications; IHC image caption). A09485 is also listed for IF and has a human brain IF image at 20 μg/mL (catalog applications; IF image caption).
Which to pick: For paraffin-section tissue IHC, choose A09485: it lists IHC-P and shows human brain staining; the fixative is unreported (catalog applications; IHC image caption). For IF, choose A09485 based on its human brain IF image; ICC validation is unreported (IF image caption; catalog applications). For mouse samples, A09485 lists mouse reactivity, but both images show human brain; its clonality is unreported (catalog reactivity; image captions; catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q92784 (DPF3_HUMAN, Zinc finger protein DPF3).
- Human Protein Atlas. DPF3 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. DPF3 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. DPF3 antibody validation summary (2 antibodies).
- The renal cancer risk allele at 14q24.2 activates a novel hypoxia-inducible transcription factor-binding enhancer of DPF3 expression. The Journal of biological chemistry 2022 — PMC8897700.
- PubMed PMID:18765789 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:12508121 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DPF3 (Zinc finger protein DPF3). Expected localisation Nuclear staining in positive tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A09485 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (GLUC cells - nucleus); 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 A09485); 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-DPF3, 5 μg/mL (datasheet A09485); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining has medium concordance with RNA data (HPA tissue IHC); Isoform / epitope: Five isoforms; epitope coverage needs confirmation (UniProt); Regulation: Staining varies by tissue and cell type (HPA tissue IHC). Evidence: UniProt Q92784, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm., Datasheet A09485, PMC8897700. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DPF3/immunohistochemistry-ihc-guide.