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HNRNPF / Heterogeneous nuclear ribonucleoprotein F · IHC design guide
Design Immunohistochemistry for HNRNPF
Plan HNRNPF paraffin IHC around nuclear staining across tissues (HPA tissue IHC). Start with the catalog antibody's 0.5–1 μg/mL IHC range (datasheet A05806) and compare staining with the tissue profile (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HNRNPF 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 | Ubiquitous nuclear staining (HPA tissue IHC) | |
| Staining pattern | Nuclei across many tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A05806) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Heart muscle |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep tissue fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining may reflect proteins from more than one gene (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | No annotated isoforms; N-terminal processing at residue 2 (UniProt) |
Section 1
Recommended HNRNPF IHC & IF Protocols
The catalog antibody uses citrate pH 6 retrieval (datasheet A05806). These published chromogenic IHC protocols cover Merkel cell carcinoma (PMC4181674) and gastric cancer (PMC10590441).
| Sample | Paraffin-embedded human mammary cancer tissues; fixative not specified (datasheet A05806) |
| Fixation | Image fixative and duration unreported (datasheet A05806); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A05806) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05806) |
| Primary antibody | Rabbit anti-HNRNPF, 0.5-1μg/ml (datasheet A05806) |
| Primary incubation | Overnight at 4 °C (datasheet A05806) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A05806) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HNRNPF-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate pH 6 retrieval (datasheet A05806); the gastric cancer protocol specifies this pH (PMC10590441), while the Merkel cell carcinoma protocol does not state a pH (PMC4181674).
Section 2
What Is the Expected HNRNPF Staining Pattern?
HNRNPF is a nucleoplasmic protein with no transmembrane segment (UniProt P52597: location and topology). Expect nuclear staining across many cell types, with intensity varying by tissue and cell type (HPA tissue IHC: ubiquitous nuclear expression). HPA rates its tissue IHC evidence Supported, with medium agreement between staining and RNA data, and cautions that the staining may include protein from more than one gene (HPA tissue IHC: reliability).
| Distinct nuclear chromogen in glandular cells, respiratory epithelium, or marrow hematopoietic cells. | This fits the expected compartment (UniProt P52597: nucleoplasm) and cell patterns (HPA tissue IHC: High in adrenal, appendix and breast glandular cells; bronchial respiratory epithelial cells; and bone marrow hematopoietic cells). Score the proportion and intensity of stained nuclei within the relevant cell population. Nuclear staining supports the expected pattern, but the HPA warning about staining protein from more than one gene limits a claim of HNRNPF specificity (HPA tissue IHC: reliability). |
| Predominantly cytoplasmic or membrane staining with little nuclear staining. | This conflicts with the expected nucleoplasmic location and lack of a transmembrane segment (UniProt P52597: location and topology; HPA subcellular: mainly nucleoplasm). Treat it as a possible artefact or nonspecific signal. Review staining controls and whether nuclei can be clearly identified before calling the slide positive. |
| Strong cardiomyocyte staining, especially without a matching nuclear pattern. | Cardiomyocytes were not detected in the supplied tissue IHC record (HPA tissue IHC: heart muscle). Consider cross-reactivity or endogenous detection activity before interpreting this as HNRNPF. Compare the subcellular pattern and a detection-only control; the HPA finding is an observed pattern, not proof that every cardiomyocyte sample must be negative. |
| Diffuse chromogen obscures nuclear boundaries or appears across the section. | A diffuse deposit cannot establish the expected nuclear pattern (UniProt P52597: nucleoplasm; HPA tissue IHC: ubiquitous nuclear expression). Evaluate detection-only background, blocking, washing and chromogen development as general IHC checks. Do not score diffuse color as positive nuclei merely because HPA describes broad tissue expression (HPA tissue IHC: profile). |
| No nuclear signal in a tissue expected to stain strongly. | Check the sampled cell population first: HPA reports High staining in several defined populations, including adrenal glandular cells and bone marrow hematopoietic cells (HPA tissue IHC: positive tissue entries). Then examine section quality, retrieval and detection controls using general IHC practice. A failed control or absent expected cells prevents a confident biological negative call. |
💡Expected HNRNPF appearanceCall the result positive when identifiable nuclei show clear chromogen in an expected cell population, often strong in HPA High populations; predominantly cytoplasmic color or diffuse background is suspect (UniProt P52597: nucleoplasm; HPA tissue IHC: High entries and ubiquitous nuclear profile).
| Cell population and expected intensity | Choose the population before scoring: HPA reports High staining in the listed glandular, respiratory epithelial, hematopoietic and selected brain cell populations, Low in hepatocytes, and Not detected in cardiomyocytes (HPA tissue IHC: positive, low and negative entries). A weak liver signal therefore carries a different expectation from a weak adrenal glandular signal. |
| IHC evidence and antibody specificity | The tissue profile is Supported, with medium agreement between staining and RNA data, and HPA cautions that the signal may include protein from more than one gene (HPA tissue IHC: reliability). HPA016884 has Supported IHC validation; HPA069667 has no IHC status in the supplied list (HPA antibodies: validation). Interpret staining as pattern evidence, with this specificity limit. |
| Retrieval and epitope information | The supplied records give no antibody epitope location or HNRNPF-specific retrieval response (UniProt P52597: supplied record; HPA tissue IHC: supplied record). If optimizing an IHC run, compare retrieval conditions against a suitable positive tissue and detection control as general IHC practice. Do not infer fixation sensitivity from topology, modifications or tissue staining levels. |
| IF/ICC Q&A: where should signal appear? | Predominantly in the nucleoplasm (HPA subcellular: mainly nucleoplasm; UniProt P52597: location). HPA lists ICC/IF images in A-431, U-251MG, U2OS, A-549 and HaCaT; HPA016884 is ICC Enhanced and HPA069667 is ICC Supported (HPA subcellular: image cell lines; HPA antibodies: validation). Those observations guide localisation, while IF/ICC methods belong in its separate guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in adrenal glandular cells or marrow hematopoietic cells. | Both are reported High, so the result may reflect a run or sampling problem (HPA tissue IHC: adrenal gland and bone marrow). | Confirm that the named cells are present, then check the positive control, retrieval record, antibody application and detection reagents. These are general IHC checks; the supplied sources do not identify an HNRNPF-specific retrieval condition. |
| Only cytoplasmic or membrane color is visible. | The compartment conflicts with nucleoplasmic HNRNPF and its lack of a transmembrane segment (UniProt P52597: location and topology). | Use the counterstain to locate nuclei; compare detection-only and positive-control sections. Reassess the staining run before scoring cells positive. |
| Cardiomyocytes show conspicuous staining. | This departs from the supplied Not detected observation; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC: cardiomyocytes). | Check whether color is truly nuclear. Inspect a detection-only control and the detection blocking steps as general chromogenic IHC practice; report the observed discrepancy if it persists. |
| Background obscures otherwise plausible nuclear staining. | Diffuse color prevents a reliable compartment call against the expected nuclear pattern (HPA tissue IHC: ubiquitous nuclear expression). | Inspect detection-only background, washing and chromogen development; adjust the run using general IHC controls, then score only nuclei whose boundaries remain distinguishable. |
| Hepatocytes stain weakly beside stronger cells. | Low hepatocyte staining is reported, so lower intensity there may fit the tissue profile (HPA tissue IHC: Low in hepatocytes). | Score hepatocytes separately from adjacent cell types and compare with a high-staining control population. Avoid treating low intensity alone as an assay failure. |
| A clean nuclear pattern is being treated as definitive HNRNPF identity. | HPA warns that its tissue staining may include protein from more than one gene (HPA tissue IHC: reliability). | Report the nuclear pattern and antibody used, including the specificity caveat. If gene-level attribution is essential, seek independent validation before making that claim. |
Experiment evidence
IHC & IF images showing HNRNPF staining
A05806 · IHCIHC analysis of HnRNPF using anti-HnRNPF antibody (A05806). HnRNPF was detected in paraffin-embedded section of human mammary cancer tissues
A05806 · IHCIHC analysis of HnRNPF using anti-HnRNPF antibody (A05806). HnRNPF was detected in paraffin-embedded section of human intestinal cancer tiss
A05806 · IFIF analysis of HnRNPF using anti-HnRNPF antibody (A05806). HnRNPF was detected in immunocytochemical section of SKOV-3 cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000169813-HNRNPF/tissue/cerebral+cortex#imid_4236764
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000169813-HNRNPF/tissue/colon#imid_4236698
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000169813-HNRNPF/tissue/liver#imid_4236683
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000169813-HNRNPF/tissue/kidney#imid_4236692
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA016884; green = HNRNPF, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA016884; green = HNRNPF, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HNRNPF IHC & IF
🧪Run bone marrow first: hematopoietic cell nuclei should stain (HPA: High in hematopoietic cells; UniProt P52597: nucleoplasm). Use heart muscle as the negative tissue (HPA: Not detected in cardiomyocytes); within marrow, mature erythrocytes, if present, should lack nuclear DAB staining because they are anucleate (standard histology).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Heart muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show HNRNPF in A-431, U-251MG, U2OS, A-549, HaCaT, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a rabbit IgG isotype control matched to the primary antibody’s clonality where applicable, and HNRNPF knockout material as a biological negative (A05806 tissue-IHC caption: rabbit primary; standard IHC practice). Block endogenous peroxidase in bone marrow and assess endogenous biotin background if using the caption’s biotin-based detection (standard IHC practice; A05806 tissue-IHC caption: biotinylated secondary and SABC).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative is unreported in the A05806 paraffin-section caption (A05806 tissue-IHC caption). That caption uses heat retrieval in citrate buffer at pH 6 for 20 minutes in mammary cancer tissue; whether bone marrow requires that condition is unreported (A05806 tissue-IHC caption). Frozen-section performance and whether IF is easier are unreported; marrow’s endogenous peroxidase can complicate chromogenic scoring (standard IHC practice), while cultured-cell ICC-IF images show a nucleoplasmic location (HPA subcellular: nucleoplasm).
HPA tissue IHC evidence for HNRNPF
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HNRNPF IHC Tips
Troubleshoot HNRNPF staining by checking nuclear localisation, retrieval conditions, antibody specificity and cell level scoring (UniProt P52597; HPA tissue IHC).
Where should I start when HNRNPF staining is weak in paraffin sections?
Start with heat mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A05806). The selected paraffin tissue example used 1 μg/mL catalog antibody overnight at 4 °C, so use those conditions as a starting point when assessing weak staining (datasheet A05806). Compare retrieved and unretrieved sections in the same run, keeping detection and development times constant (standard IHC practice). If staining remains weak, test retrieval time on adjacent sections while watching for tissue damage and nonspecific DAB deposition (standard IHC practice). The example does not report its fixative, so it cannot establish fixation dependent retrieval requirements (datasheet A05806).
Can I choose a fixative based on the published HNRNPF staining pattern?
Target specific fixation sensitivity is unknown: the selected paraffin section caption does not state a fixative (datasheet A05806). Its citrate pH 6 retrieval for 20 minutes establishes a reported processing step, but does not identify which fixation conditions preserve the epitope (datasheet A05806). Record fixative, fixation duration and processing history for each specimen, then compare adjacent sections using the same retrieval, antibody concentration and DAB development (standard IHC practice). Include a nuclear positive control and a no primary control to distinguish lost signal from detection background (UniProt P52597 localisation; standard IHC practice). Do not infer fixation tolerance from HPA tissue staining (HPA tissue IHC).
What HNRNPF staining pattern should I expect in a positive section?
Score nuclear staining, with emphasis on the nucleoplasm: UniProt assigns HNRNPF to the nucleoplasm and HPA reports predominantly nucleoplasmic staining (UniProt P52597 localisation; HPA subcellular). HNRNPF has no transmembrane segment, so a crisp membrane rim is unexpected for this target (UniProt P52597 topology). Use a nuclear counterstain to confirm that DAB falls within intact nuclei rather than over cytoplasm or extracellular material (standard IHC practice). HPA describes ubiquitous nuclear expression, with high staining in breast glandular cells and undetected staining in heart cardiomyocytes among its examples (HPA tissue IHC). Treat those examples as context because HPA cautions that its staining can reflect proteins from more than one gene (HPA tissue IHC).
Could processing or an unmapped epitope explain discordant HNRNPF staining?
The supplied UniProt record lists 0 isoforms but describes full length 1–415 and N terminally processed 2–415 chains (UniProt P52597 processing). It also places RNA recognition motifs at 13–85, 111–188 and 289–366, with reported phosphorylation at residues including 104, 107 and 161 (UniProt P52597 domains and modified residues). The antibody epitope is not specified in the supplied caption, so these annotations cannot identify which modification or processing event affects staining (datasheet A05806; UniProt P52597). If sections disagree, first compare processing and retrieval records, then verify antibody specificity with an appropriate independent control (standard IHC practice). Interpret discordant nuclear signals cautiously given HPA's cross gene staining warning (HPA tissue IHC).
How can I check nuclear HNRNPF by multiplex IF alongside this IHC assay?
Use the separate IF/ICC workflow to examine nucleoplasmic signal, the expected location for HNRNPF (UniProt P52597 localisation; HPA subcellular). Pair HNRNPF with a marker of the cell type being assessed and a nuclear counterstain, then inspect whether signals occupy the intended cells and nuclei (standard IF practice). Choose a fluorophore whose emission is distinguishable from the specimen's autofluorescence, and include single stain controls when setting channels (standard IF practice). Because HNRNPF is nucleoplasmic and has no transmembrane segment, permeabilisation must allow antibody access inside the nucleus; optimise it against cell preservation (UniProt P52597 topology and localisation; standard IF practice). Do not transfer the paraffin section's fixation assumptions to IF (datasheet A05806).
What should I change if DAB obscures HNRNPF positive nuclei?
Run a no primary control and examine whether brown signal remains in tissue compartments or at section edges (standard IHC practice). For chromogenic detection, block endogenous peroxidase before DAB development and titrate development time against a matched positive section (standard IHC practice). The reported example used 10% goat serum blocking, 1 μg/mL catalog antibody overnight at 4 °C, and a biotin based DAB detection sequence (datasheet A05806). If background persists, assess whether the biotin based detection system contributes signal by comparing an alternative detection system (datasheet A05806; standard IHC practice). Accept signal only when it resolves within nuclei, consistent with HNRNPF localisation (UniProt P52597 localisation).
How should I score HNRNPF when most cells show some nuclear signal? ⚠ ANSWER MARKED FOR VERIFICATION
Define the analysed cell population and score nuclear DAB signal only, using the counterstain to identify intact nuclei (UniProt P52597 localisation; standard IHC practice). Report the percentage of positive nuclei and, when intensity matters, an H-score calculated from percentages at each prespecified intensity level (standard IHC practice). For spatial comparisons, positive nuclei per mm² can supplement that score if the analysed area is recorded (standard IHC practice). Normalise positive counts to all eligible nuclei in the same cell population, and keep retrieval, exposure to detection reagents and DAB development consistent across sections (standard IHC practice). HPA's ubiquitous nuclear profile makes a declared positivity threshold especially useful (HPA tissue IHC).
How do I distinguish genuine HNRNPF positivity from staining artefacts?
A convincing result places DAB within intact nuclei, particularly the nucleoplasm, in the cells being evaluated (UniProt P52597 localisation; HPA subcellular). Treat isolated membrane or extracellular staining as discordant with the recorded location, and check whether signal concentrates at section edges or necrotic areas (UniProt P52597 topology and localisation; standard IHC practice). Compare those areas with a no primary control and with endogenous peroxidase blocked sections before attributing brown deposits to HNRNPF (standard IHC practice). HPA reports high staining in several cell populations but also warns that its antibody staining can reflect proteins from more than one gene (HPA tissue IHC). Confirm consequential findings with an independent specificity control (standard IHC practice).
Boster reagents
Best HNRNPF / Heterogeneous nuclear ribonucleoprotein F IHC Antibodies
A05806 has IHC images from paraffin sections of human mammary and intestinal cancers and mouse and rat intestine (catalog IHC captions), plus IF data in SKOV-3 cells (catalog IF caption).
Real IHC data
Anti-HnRNP F/HNRNPF Antibody ®
Cat # A05806
A05806 is listed for IHC and has images from human mammary cancer, human intestinal cancer, mouse intestine, and rat intestine paraffin sections (catalog applications; catalog IHC captions). A05806 is also listed for IF and ICC, with an IF image from SKOV-3 cells (catalog applications; catalog IF caption).
Which to pick: Choose A05806 for paraffin-section IHC because its own captions document that preparation across the listed human, mouse, and rat samples (catalog IHC captions). Choose A05806 for IF/ICC because those applications are listed and its IF caption documents staining in SKOV-3 cells (catalog applications; catalog IF caption). For cross-species IHC, A05806 lists human, mouse, and rat reactivity and shows IHC images from all three; it is rabbit-hosted, its clonality is unreported, and the IHC captions do not report the fixative (catalog reactivity; catalog IHC captions; catalog host and clone fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P52597 (HNRPF_HUMAN, Heterogeneous nuclear ribonucleoprotein F).
- Human Protein Atlas. HNRNPF tissue IHC expression (reliability: Supported).
- Human Protein Atlas. HNRNPF subcellular location (ICC-IF): Mainly localized to the nucleoplasm..
- Human Protein Atlas. HNRNPF antibody validation summary (2 antibodies).
- A Proteomic Study of Human Merkel Cell Carcinoma. Journal of proteomics & bioinformatics 2013 — PMC4181674.
- The VAX2-LINC01189-hnRNPF signaling axis regulates cell invasion and migration in gastric cancer. Cell death discovery 2023 — PMC10590441.
- PubMed PMID:7512260 — UniProt-cited evidence.
- PubMed PMID:7499401 — UniProt-cited evidence.
- PubMed PMID:1351868 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HNRNPF (Heterogeneous nuclear ribonucleoprotein F). Expected localisation Ubiquitous nuclear staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A05806). Validated IHC antibody: A05806 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Heart muscle. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human mammary cancer tissues; fixative not specified (datasheet A05806); 2. Fixation: Image fixative and duration unreported (datasheet A05806); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: Citrate pH 6 , 20 min (datasheet A05806); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05806); 8. Primary antibody: Rabbit anti-HNRNPF, 0.5-1μg/ml (datasheet A05806); 9. Primary incubation: Overnight at 4 °C (datasheet A05806). Watch for: Fixation: Keep tissue fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining may reflect proteins from more than one gene (HPA tissue IHC); Isoform / epitope: No annotated isoforms; N-terminal processing at residue 2 (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). Evidence: UniProt P52597, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm., Datasheet A05806, PMC4181674, PMC10590441. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HNRNPF/immunohistochemistry-ihc-guide.