This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
HCFC1 / Host cell factor 1 · IHC design guide
Design Immunohistochemistry for HCFC1
Plan HCFC1 paraffin IHC with the catalog antibody at 2–5 μg/ml (datasheet A01729-2) and assess nuclear and cytoplasmic staining (HPA tissue IHC). Use highly stained nasopharyngeal ciliated cells as a positive reference (HPA tissue IHC), and consider HCFC1R1-dependent redistribution when interpreting compartment shifts (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HCFC1 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 and cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Variable nuclear and cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01729-2) | |
| Positive control | Nasopharynx+4 more · see all | |
| Negative control | Adrenal gland+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 | HCFC1R1 overexpression can shift HCFC1 to cytoplasm (UniProt) | |
| Regulation | Higher expression in fetal tissues (UniProt) | |
| Isoform / epitope | 4 isoforms; N/C cleavage makes epitope location relevant (UniProt) |
Section 1
Recommended HCFC1 IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 heat retrieval (datasheet A01729-2). One published HCFC1 IHC protocol describes staining FFPE hepatocellular carcinoma specimens (PMC10240668).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A01729-2) |
| Fixation | Image fixative and duration unreported (datasheet A01729-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A01729-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01729-2) |
| Primary antibody | Rabbit anti-HCFC1, 2-5 μg/ml (datasheet A01729-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01729-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01729-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HCFC1-positive staining in ciliated cells (cell body) of nasopharynx (HPA tissue IHC: High). HPA tissue profile: Nuclear and cytoplasmic expression in varying fraction cells in most tissues. No signal in the no-primary control. |
💡Decision noteTry EDTA pH 8.0 heat retrieval first (datasheet A01729-2); the published protocol does not specify retrieval conditions (PMC10240668).
Section 2
What Is the Expected HCFC1 Staining Pattern?
HCFC1 is found in the nucleus and cytoplasm, with no transmembrane segment (UniProt P51610). In paraffin-section IHC, expect staining in a varying fraction of cells across most tissues (HPA tissue IHC: Supported; medium consistency with RNA). Nasopharyngeal ciliated cell bodies provide a high-staining example; bronchial ciliated cells and lung macrophages are medium-staining examples (HPA tissue IHC).
| Nuclear and cytoplasmic staining appears in a varying fraction of cells within a tissue (HPA tissue IHC). | This matches the reported tissue pattern. Assess nuclei and cell bodies separately, and record the fraction stained; uniform staining of every cell is not required by the HPA observation (HPA tissue IHC). |
| Strong signal outlines cell membranes or fills extracellular spaces, with little nuclear or cell-body staining. | Treat this as suspect compartmental staining: HCFC1 is nuclear and cytoplasmic and has no transmembrane segment (UniProt P51610). Review morphology, antibody controls and detection background before interpreting it as HCFC1. |
| Prominent staining occurs in adrenal gland glandular cells or lymph-node germinal-center cells. | These specific cell populations were not detected in HPA tissue IHC. Check whether the signal follows the expected intracellular pattern, and consider cross-reactivity or endogenous detection activity; the HPA finding does not make the entire tissue a negative control. |
| Chromogen appears broadly across tissue, including areas without recognizable cell boundaries. | A diffuse field cannot establish the reported cellular pattern (HPA tissue IHC). In general IHC practice, inspect the no-primary control and detection reagents for background before assigning positive cells or scoring intensity. |
| Nasopharyngeal ciliated cell bodies show no convincing signal. | This conflicts with HPA's high-staining example (HPA tissue IHC). Confirm that the relevant cells are present, then review the catalog antibody's IHC-P conditions and assay controls; one absent signal does not by itself establish lack of HCFC1. |
💡Expected HCFC1 appearanceCall a section positive when identifiable cells show nuclear and/or cell-body cytoplasmic staining, with high signal possible in nasopharyngeal ciliated cell bodies (HPA tissue IHC); diffuse, cell-free chromogen or isolated membrane outlines are suspect against the reported localization (HPA tissue IHC; UniProt P51610).
| Tissue and cell selection | HPA reports high staining in nasopharyngeal ciliated cell bodies; bronchial ciliated cells, lung macrophages and placental trophoblastic cells are medium (HPA tissue IHC). Compare the named cell population, since a tissue-wide average can hide a focal result. |
| Strength of the tissue evidence | The tissue IHC pattern is Supported, with medium consistency between antibody staining and RNA expression (HPA tissue IHC). Treat a surprising result as a reason to review controls and morphology, rather than as a definitive biological exception. |
| Subcellular context | Tissue IHC reports nuclear and cytoplasmic expression (HPA tissue IHC); ICC-IF supports nucleoplasm as the main location (HPA subcellular). UniProt notes that HCFC1R1 overexpression can accumulate HCFC1 in cytoplasm (UniProt P51610), but this does not establish a shift in a given tissue. |
| Processing and epitope coverage | HCFC1 produces associated N- and C-terminal cleavage products (UniProt P51610). The supplied antibody record gives no epitope, so staining cannot identify which chain or cleavage state the catalog antibody recognizes. |
| Isoforms and antibody validation | Four isoforms are listed (UniProt P51610), while HPA078588 is Supported for both IHC and ICC (HPA antibodies). Neither entry establishes equal recognition of all isoforms or transfers an ICC-IF image result to paraffin sections. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal appears in the expected nasopharyngeal ciliated cells (HPA tissue IHC). | The scored section may lack intact target cells, or the IHC run may have failed; the image alone cannot distinguish these possibilities. | Confirm cell identity and preservation, then review the catalog antibody's IHC-P instructions, reagent performance and a concurrent positive control. General IHC practice: change one workflow variable at a time. |
| Only membrane outlines or extracellular material stain. | That distribution is difficult to reconcile with nuclear and cytoplasmic HCFC1 and its lack of a transmembrane segment (UniProt P51610). | Compare with a no-primary control and examine whether chromogen tracks tissue edges or damaged areas. General IHC practice: resolve detection background before interpreting compartment-specific signal. |
| Unexpected cells stain strongly while expected cells remain faint. | The named HPA-negative populations are cell-specific observations, and unexpected signal may reflect cross-reactivity or endogenous detection activity (HPA tissue IHC; general IHC practice). | Identify the stained cells on the counterstain; compare their compartment and morphology with a known positive example, then inspect no-primary and detection controls. |
| A broad, weak haze obscures nuclei and cell bodies. | Diffuse background prevents a reliable call on the nuclear and cytoplasmic pattern reported by HPA (HPA tissue IHC). | General IHC practice: inspect the no-primary control, blocking and washes, and score only discrete cellular staining after the background is controlled. |
| Cytoplasmic staining exceeds nuclear staining in an otherwise interpretable section. | Cytoplasmic HCFC1 is reported in tissue IHC, and HCFC1R1 overexpression can alter localization (HPA tissue IHC; UniProt P51610). Neither source explains a particular specimen's balance. | Record nuclear and cytoplasmic intensity separately, check a comparable section and controls, and avoid inferring HCFC1R1 status from staining alone. |
| What should an IF/ICC image show? | HPA identifies nucleoplasm as the supported main ICC-IF location and lists A-549, K-562 and U2OS images (HPA subcellular). | Look for nucleoplasmic signal when interpreting ICC-IF (HPA subcellular). Use the separate IF/ICC guide for that application; the HPA image result alone does not specify an IHC-P protocol. |
Experiment evidence
IHC & IF images showing HCFC1 staining
A01729-2 · IHCIHC analysis of HCFC1 using anti-HCFC1 antibody (A01729-2). HCFC1 was detected in a paraffin-embedded section of human breast cancer tissue.
A01729-2 · IHCIHC analysis of HCFC1 using anti-HCFC1 antibody (A01729-2). HCFC1 was detected in a paraffin-embedded section of human liver cancer tissue.
A01729-2 · IFIF analysis of HCFC1 using anti-HCFC1 antibody (A01729-2) and anti-Beta Tubulin antibody (M01857-3). HCFC1 was detected in an immunocytochem
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000172534-HCFC1/tissue/cerebral+cortex#imid_20823412
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000172534-HCFC1/tissue/colon#imid_20822754
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000172534-HCFC1/tissue/liver#imid_20822763
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000172534-HCFC1/tissue/kidney#imid_20822764
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA078588; green = HCFC1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA078588; green = HCFC1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HCFC1 IHC & IF
🧪Run nasopharynx first and look for staining in ciliated cell bodies (HPA: nasopharynx ciliated cells, High). Use adrenal gland glandular cells as a low-signal comparison (HPA: adrenal gland glandular cells, Not detected); on the nasopharynx slide, compare the ciliated cell bodies with neighboring cells that lack visible staining, without treating those neighbors as a validated biological negative.
Positive control tissue: Nasopharynx (Ciliated cells (cell body), HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show HCFC1 in A-549, K-562, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (caption: rabbit primary and anti-rabbit secondary); a validated HCFC1 knockout specimen can test target specificity (standard IHC practice). Block endogenous peroxidase and check for background on the nasopharynx section before interpreting DAB staining (caption: peroxidase detection with DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A01729-2 paraffin-section caption does not state a fixative (caption: fixative not stated). Heat-mediated retrieval in EDTA, pH 8.0 was used, but retrieval dependency was not established (caption: EDTA retrieval). The supplied evidence does not establish whether frozen sections or IF would be easier; for nasopharyngeal mucosa, check background from endogenous peroxidase when scoring the ciliated cell bodies (HPA: nasopharynx ciliated cells, High; caption: peroxidase detection with DAB; standard IHC practice).
HPA tissue IHC evidence for HCFC1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium 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 |
|---|---|---|---|---|
| Nasopharynx | Ciliated cells (cell body) | High | Protein (IHC) | HPA → |
| Bronchus | Ciliated cells (cell body) | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (cell body) | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Colon | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Peripheral nerve | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HCFC1 IHC Tips
These questions address HCFC1 staining in paraffin sections with chromogenic IHC detection, with one section on IF/ICC.
How should I optimize retrieval when HCFC1 staining is weak?
Start with heat-mediated EDTA retrieval at pH 8.0 for paraffin-section HCFC1 IHC (datasheet A01729-2). The selected tissue image used this condition before 2 μg/ml primary antibody incubation overnight at 4°C (datasheet A01729-2). If staining is weak, verify deparaffinisation and consistent heating and cooling, then vary retrieval duration on adjacent sections while holding detection constant (standard IHC practice). If that fails, compare another validated buffer and pH as a fallback, recording each condition rather than assuming the epitope is accessible (standard IHC practice). Assess nuclear and cytoplasmic staining separately because both occur in tissue IHC (HPA tissue IHC).
Could fixation explain inconsistent HCFC1 staining between sections?
The selected paraffin-section caption does not report a fixative, so HCFC1-specific fixation sensitivity is unknown (datasheet A01729-2). Neither the tissue staining pattern nor HCFC1 topology establishes how a particular fixative affects its epitope (HPA tissue IHC; UniProt P51610 topology). For prospectively prepared specimens, record fixative identity, concentration, tissue thickness, and fixation time, and keep them consistent across comparisons (standard IHC practice). If batches differ, stain adjacent control sections with matched retrieval and detection, changing one preparation variable at a time (standard IHC practice). Evaluate signal alongside morphology and background, with particular attention to the expected nuclear compartment (HPA subcellular; standard IHC practice).
Should HCFC1 staining be nuclear or cytoplasmic in tissue sections?
Expect prominent nuclear staining, especially nucleoplasmic signal, while allowing cytoplasmic staining in some tissue cells (HPA subcellular; HPA tissue IHC). HCFC1 is annotated in both nucleus and cytoplasm, and excess HCFC1R1 can shift it toward cytoplasmic accumulation (UniProt P51610 subcellular, PubMed:12235138). Score these compartments separately instead of treating diffuse whole-cell color as a single result (standard IHC practice). Compare staining with recognizable cell boundaries and a nuclear counterstain, because dense chromogen can obscure the distinction between nuclear and adjacent cytoplasmic signal (standard IHC practice). A predominantly cytoplasmic result warrants checking controls and processing before assigning a biological redistribution (HPA subcellular; standard IHC practice).
Can this IHC antibody distinguish HCFC1 isoforms or processed chains?
HCFC1 has 4 annotated isoforms and is processed into multiple N-terminal and C-terminal chains (UniProt P51610 isoforms and processing). Its cleavage products generally remain tightly associated, so their presence cannot be distinguished merely by observing a shared cellular staining pattern (UniProt P51610 subunit). The selected IHC caption does not identify the antibody epitope or demonstrate isoform selectivity (datasheet A01729-2). Request an epitope map and check which isoforms and processed chains retain that sequence before making a chain-specific claim (UniProt P51610 processing; standard IHC practice). Until specificity is established, report HCFC1 staining by compartment and cell type rather than attributing it to an individual isoform (standard IHC practice).
How can I assess HCFC1 localisation by IF/ICC alongside cell identity?
For IF/ICC, pair HCFC1 with a validated marker of the expected cell type; ciliated cells are one documented positive population (HPA tissue IHC: nasopharynx ciliated cells). Include a nuclear counterstain so nucleoplasmic signal can be distinguished from cytoplasmic signal in each cell (HPA subcellular; standard IF practice). Choose a spectrally separated fluorophore, favoring a longer-wavelength channel when the specimen has strong short-wavelength autofluorescence (standard IF practice). HCFC1 has no transmembrane segment and is found intracellularly, so use permeabilisation to reach the epitope, then optimize its strength because the antibody's epitope is unspecified (UniProt P51610 topology and subcellular; datasheet A01729-2; standard IF practice). Do not carry the paraffin IHC antibody concentration into IF without testing (datasheet A01729-2; standard IF practice).
What should I check when HCFC1 DAB staining appears diffuse?
The selected paraffin-section method used a peroxidase-conjugated secondary and DAB detection after 10% goat-serum blocking (datasheet A01729-2). For widespread color, inspect a no-primary control and block endogenous peroxidase before DAB development to identify detection-related signal (standard IHC practice). Check whether secondary binding, inadequate washing, or excessive development produces similar staining outside recognizable cells (standard IHC practice). Compare intact tissue with edges and damaged areas, where uneven processing can complicate interpretation (standard IHC practice). Retain genuine nuclear and cytoplasmic signal as possibilities, because both compartments occur in HCFC1 tissue IHC, but require cell-resolved staining above control background (HPA tissue IHC; standard IHC practice).
How should I quantify HCFC1 across heterogeneous tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue region and cell population before scoring, because HCFC1 appears in varying fractions of cells across tissues (HPA tissue IHC; standard IHC practice). Record the percentage of positive cells and nuclear and cytoplasmic intensity separately; an H-score can combine intensity categories 0–3 with their cell percentages (standard IHC practice). For spatial comparisons, report positive-cell density per mm² of evaluable tissue and exclude folds, necrosis, and poorly preserved regions by a preset rule (standard IHC practice). Normalize counts to the relevant cell population or evaluable area, and apply one threshold and staining batch controls across specimens (standard IHC practice). Report compartment-specific results because nucleoplasmic localisation is supported while tissue IHC can also show cytoplasmic expression (HPA subcellular; HPA tissue IHC).
How can I distinguish credible HCFC1 staining from artefact?
A credible result resolves individual cells and includes nuclear signal, consistent with supported nucleoplasmic localisation; cytoplasmic staining can also occur (HPA subcellular; HPA tissue IHC). Compare cell identity with the tissue context: nasopharyngeal ciliated cells show high staining, whereas adrenal glandular cells are reported as not detected (HPA tissue IHC). Treat signal confined to cut edges, folds, or necrotic regions cautiously and inspect intact areas under the same exposure and scoring rules (standard IHC practice). Check a no-primary control and endogenous peroxidase block when DAB appears in unexpected compartments or cell populations (standard IHC practice). HPA rates tissue staining as supported with medium RNA concordance, so confirm a surprising pattern with independent evidence before assigning biological meaning (HPA tissue IHC).
Boster reagents
Best HCFC1 / Host cell factor 1 IHC Antibodies
A01729-2 has IHC images from human breast and liver cancer sections and mouse and rat liver sections (catalog IHC captions), plus IF data from U2OS cells (catalog IF caption).
Real IHC data
Anti-HCFC1 Antibody ®
Cat # A01729-2
A01729-2 is shown in paraffin sections of human breast cancer, human liver cancer, mouse liver and rat liver (catalog IHC captions). The same SKU is shown in U2OS cells by IF (catalog IF caption).
Which to pick: For tissue IHC, choose A01729-2: its images document paraffin sections with EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml (catalog IHC captions); the fixative is unreported (catalog IHC captions). For IF/ICC, A01729-2 has a U2OS cell image using 5 μg/ml primary antibody (catalog IF caption). For cross-species IHC, A01729-2 has images from human, mouse and rat tissue (catalog IHC captions); its IF reactivity is listed for human samples (catalog application data).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P51610 (HCFC1_HUMAN, Host cell factor 1).
- Human Protein Atlas. HCFC1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. HCFC1 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. HCFC1 antibody validation summary (1 antibodies).
- Hcfc1a regulates neural precursor proliferation and asxl1 expression in the developing brain. BMC neuroscience 2020 — PMC7288482.
- Upregulation of HCFC1 expression promoted hepatocellular carcinoma progression through inhibiting cell cycle arrest and correlated with immune infiltration. Journal of Cancer 2023 — PMC10240668.
- Hcfc1b, a zebrafish ortholog of HCFC1, regulates craniofacial development by modulating mmachc expression. Developmental biology 2014 — PMC4391465.
- Delivery of Multifunctional Microspheres via Intravesical Instillation for Bladder Carcinoma: Therapeutic Potential and Mechanistic Insights. Advanced healthcare materials 2026 — PMC13005678.
- PubMed PMID:8392914 — UniProt-cited evidence.
- PubMed PMID:7876203 — UniProt-cited evidence.
- PubMed PMID:15772651 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HCFC1 (Host cell factor 1). Expected localisation Nuclear and cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01729-2). Validated IHC antibody: A01729-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Nasopharynx (Ciliated cells (cell body)); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A01729-2); 2. Fixation: Image fixative and duration unreported (datasheet A01729-2); 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: EDTA pH 8.0 (datasheet A01729-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01729-2); 8. Primary antibody: Rabbit anti-HCFC1, 2-5 μg/ml (datasheet A01729-2); 9. Primary incubation: Overnight at 4 °C (datasheet A01729-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: HCFC1R1 overexpression can shift HCFC1 to cytoplasm (UniProt); Isoform / epitope: 4 isoforms; N/C cleavage makes epitope location relevant (UniProt); Regulation: Higher expression in fetal tissues (UniProt). Evidence: UniProt P51610, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm., Datasheet A01729-2, PMC10240668. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HCFC1/immunohistochemistry-ihc-guide.