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HNRNPC / Heterogeneous nuclear ribonucleoproteins C1/C2 · IHC design guide
Design Immunohistochemistry for HNRNPC
Plan HNRNPC staining in paraffin sections using the observed general nuclear pattern as a reference (HPA tissue IHC). Compare cell types within each section, including fibroblasts, which show low staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HNRNPC 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 observed (HPA tissue IHC) | |
| Staining pattern | General nuclear staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02726-1) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Fibroblasts may show low staining (HPA tissue IHC) | |
| Regulation | Regulation not annotated (UniProt) | |
| Isoform / epitope | 4 isoforms; check epitope before isoform claims (UniProt) |
Section 1
Recommended HNRNPC IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with published HNRNPC IHC protocols for lung adenocarcinoma, glioma, and colorectal cancer tissues (PMC7868529; PMC7578363; PMC12802253).
| Sample | Paraffin-embedded human bladder urothelial carcinoma tissue; fixative not specified (datasheet A02726-1) |
| Fixation | Image fixative and duration unreported (datasheet A02726-1); 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 A02726-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02726-1) |
| Primary antibody | Rabbit anti-HNRNPC, 1:50 recommended; image 1:100 (datasheet A02726-1) |
| Primary incubation | Overnight at 4 °C (datasheet A02726-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02726-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HNRNPC-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A02726-1); citrate retrieval with high-pressure heating was used for colorectal cancer sections (PMC12802253).
Section 2
What Is the Expected HNRNPC Staining Pattern?
HNRNPC is a nuclear protein with no transmembrane segment (UniProt P07910: location and topology). In paraffin tissue sections, expect nuclear staining across many cell types, including adipocytes, glandular cells, hematopoietic cells and respiratory epithelial cells (HPA: general nuclear expression; High in the listed cells). HPA rates the tissue IHC pattern Supported, citing consistency between antibody staining and RNA expression data (HPA: reliability).
| Distinct nuclear stain in adipocytes, glandular cells or hematopoietic cells. | This matches the expected compartment and documented High staining in these cell types (UniProt P07910: nucleus; HPA: tissue IHC). Compare nuclei with the counterstain when judging localisation (general IHC practice). |
| Stain chiefly outlines cell membranes or fills cytoplasm while nuclei remain clear. | That conflicts with nuclear localisation and general nuclear tissue staining (UniProt P07910: nucleus, no transmembrane segment; HPA: tissue IHC). Treat it as possible artefact and check detection controls before assigning it to HNRNPC (general IHC practice). |
| Strong staining appears in an unexpected cell population, especially fibroblasts. | HPA reports Low staining in soft tissue fibroblasts, so prominent signal there warrants review (HPA: tissue IHC). Cross-reactivity or endogenous detection activity are possibilities, not diagnoses; compare cell identity and a detection control (general IHC practice). |
| Brown precipitate is diffuse across tissue, stroma and slide background. | That distribution obscures the expected nuclear pattern (HPA: general nuclear expression). Nonspecific antibody binding or detection background may contribute; inspect a control without primary antibody and the counterstained morphology (general IHC practice). |
| A section containing documented High-staining cells has no nuclear signal. | Check the control tissue, antibody incubation, retrieval and detection run before calling the specimen negative (HPA: High in listed cell types; general IHC practice). HPA's tissue profile does not establish HNRNPC-specific retrieval or fixation sensitivity (HPA: tissue IHC scope). |
💡Expected HNRNPC appearanceCall a result positive when recognisable cell nuclei stain clearly above local background in documented High-staining populations; dominant membrane, cytoplasmic or diffuse deposit is suspect (UniProt P07910: nucleus, no transmembrane segment; HPA: tissue IHC; general IHC practice).
| Subcellular location | HNRNPC is nuclear and a component of ribonucleosomes (UniProt P07910: location). Score nuclear signal against the counterstain; a membrane-only pattern has no support from the supplied localisation evidence (UniProt P07910: topology; general IHC practice). |
| Cell and tissue context | HPA describes general nuclear expression and Low tissue RNA specificity, with High staining in several listed cell populations (HPA: tissue IHC). Soft tissue fibroblasts are listed as Low, not negative; intensity comparisons need that distinction (HPA: tissue IHC). |
| Isoforms and antigen coverage | UniProt lists four HNRNPC isoforms, including C1 and C2 (UniProt P07910: isoforms). The payload gives no antibody epitope, so it cannot establish which isoforms an IHC signal detects or predict isoform-specific staining (UniProt P07910: isoforms; supplied antibody data). |
| Antigen retrieval | Retrieval is a general paraffin IHC workflow variable (general IHC practice). No supplied source reports an HNRNPC-specific retrieval condition or fixation effect; adjust retrieval only against appropriate controls and avoid interpreting a change as proof of target-specific masking (source scope). |
| IHC antibody evidence | HPA marks HPA051075, CAB005223, CAB075755 and CAB075756 as Supported for IHC (HPA: antibodies). This supports use of their reported staining patterns, but the supplied status alone does not identify an epitope, dilution or cause of unexpected staining (HPA: antibodies). |
| IF/ICC Q&A | Q: Where should HNRNPC appear in IF/ICC? A: Predominantly in the nucleoplasm (HPA: subcellular ICC-IF). That observation helps assess compartment agreement; IF/ICC assay setup belongs to its separate guide (HPA: subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| No nuclear stain in a documented High-staining population. | The run may have failed at an antibody, retrieval or detection step; absence alone does not identify which step (HPA: tissue IHC; general IHC practice). | Confirm the documented cell population is present, then review the run control and each workflow step (HPA: tissue IHC; general IHC practice). |
| Only cytoplasmic or membrane staining is visible. | The compartment conflicts with UniProt nuclear localisation and HPA's tissue pattern (UniProt P07910: location and topology; HPA: tissue IHC). | Recheck nuclear boundaries with the counterstain and compare a control without primary antibody (general IHC practice). |
| Soft tissue fibroblasts stain more strongly than expected. | HPA lists these cells as Low; cross-reactivity or endogenous detection activity may contribute, but neither follows from the image alone (HPA: tissue IHC; general IHC practice). | Verify cell identity and inspect a detection control before attributing the strong signal to HNRNPC (general IHC practice). |
| Diffuse brown background makes nuclei hard to score. | Nonspecific binding or detection background can obscure compartment assessment (general IHC practice). | Inspect a control without primary antibody, then review blocking, antibody concentration and detection development using the assay instructions (general IHC practice). |
| One tissue appears weaker than another. | HPA reports a broad pattern with differing listed staining levels; a weaker field is not automatically a technical failure (HPA: tissue IHC). | Compare the same identified cell types and scoring criteria, and check a documented High-staining control from the run (HPA: tissue IHC; general IHC practice). |
| IF/ICC localisation seems inconsistent with tissue IHC. | HPA reports nucleoplasmic ICC-IF localisation and general nuclear tissue IHC; the payload does not establish an assay-specific biological difference (HPA: subcellular ICC-IF and tissue IHC). | Check whether each image actually resolves nuclei before interpreting the discrepancy; follow the separate IF/ICC guide for that assay (general microscopy practice). |
Experiment evidence
IHC & IF images showing HNRNPC staining
A02726-1 · IHCIHC analysis of HNRNPC using anti-HNRNPC antibody (A02726-1). HNRNPC was detected in a paraffin-embedded section of human bladder urothelial
A02726-1 · IHCIHC analysis of HNRNPC using anti-HNRNPC antibody (A02726-1). HNRNPC was detected in a paraffin-embedded section of human breast cancer tiss
A02726-1 · IFIF analysis of HNRNPC using anti-HNRNPC antibody (A02726-1) and anti-Tubulin Alpha antibody (M03989-3). HNRNPC was detected in immunocytoche
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000092199-HNRNPC/tissue/cerebral+cortex#imid_15367927
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000092199-HNRNPC/tissue/colon#imid_15367853
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000092199-HNRNPC/tissue/liver#imid_1583019
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000092199-HNRNPC/tissue/kidney#imid_1583020
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA051075; green = HNRNPC, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA051075; green = HNRNPC, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HNRNPC IHC & IF
🧪Run adipose tissue first and look for nuclear staining in adipocytes (HPA: High in adipocytes; UniProt P07910: nucleus). HPA detects HNRNPC in all 45 scored tissues, so use no-primary and isotype controls for the negative comparison; no internal target-negative cell type is established, and any unstained neighboring cells should have intact counterstained nuclei before being interpreted as negative (HPA: no negative tissue rows; standard IHC practice).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: HNRNPC is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show HNRNPC in HEK293, U-251MG, U2OS, A-431, HAP1, K-562, U2OS, siRNA 1 (10x), U2OS, siRNA 1 (40x), U2OS, siRNA 2 (10x), U2OS, siRNA 2 (40x), U2OS, scrambled (10x), U2OS, scrambled (40x), with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section, a concentration-matched rabbit isotype control, and HNRNPC-knockout material as a biological negative (selected-SKU caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase in the adipose section before HRP/DAB detection (selected-SKU caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption does not state its fixative (selected-SKU caption: fixative unreported). Heat retrieval with EDTA at pH 8.0 and a 1:100 primary dilution were reported for a bladder carcinoma section; their performance in adipose tissue is unreported (selected-SKU caption). Frozen sections have no supplied comparison, while ICC-IF images show nucleoplasmic localization; in adipose sections, the thin rim of cytoplasm and displaced adipocyte nucleus can make nuclear scoring difficult (HPA: nucleoplasm enhanced; standard adipose histology).
HPA tissue IHC evidence for HNRNPC
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — High 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: HNRNPC is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced HNRNPC IHC Tips
Troubleshoot HNRNPC staining in paraffin sections by checking nuclear localisation, retrieval, detection background and the cell population being scored (UniProt P07910; HPA tissue IHC).
What should I change when nuclear HNRNPC staining is weak after retrieval?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A02726-1). The documented example used a 1:100 primary dilution overnight at 4°C, so match those conditions before changing retrieval (datasheet A02726-1). If nuclear staining remains weak, adjust heating time in small increments on matched sections and monitor tissue integrity (standard IHC practice). Compare each condition with a no-primary control and a section containing readily identifiable nuclei, because HNRNPC is expected in nuclei (standard IHC practice; UniProt P07910 localisation). Record the retrieval time and cooling conditions for every slide to make the comparison interpretable (standard IHC practice).
Could fixation explain variable HNRNPC staining across paraffin sections?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (datasheet A02726-1). Record each specimen’s fixative, fixation duration and processing history before attributing staining differences to HNRNPC abundance (standard IHC practice). Compare sections with similar processing, using the documented EDTA pH 8.0 retrieval and 1:100 overnight primary incubation as a common starting point (datasheet A02726-1). If staining varies, inspect nuclear morphology and repeat the comparison with matched control tissue and a no-primary slide (standard IHC practice). Neither the nuclear localisation nor the listed modifications establish a fixation effect (UniProt P07910 localisation and modified residues).
Where should I expect HNRNPC staining, and how should cytoplasmic signal be assessed?
Expect predominantly nuclear staining: UniProt places HNRNPC in the nucleus, and HPA reports enhanced nucleoplasmic localisation and general nuclear tissue expression (UniProt P07910 localisation; HPA subcellular; HPA tissue IHC). Score tumour and adjacent cell populations separately when their nuclear staining differs (standard IHC practice). If cytoplasmic colour dominates, check the no-primary control, chromogen development and section morphology before treating it as HNRNPC localisation (standard IHC practice; UniProt P07910 localisation). HNRNPC has no transmembrane segment, so a crisp membrane pattern also warrants scrutiny (UniProt P07910 topology). Use the counterstain to confirm that the measured signal overlaps intact nuclei (standard IHC practice).
Can this IHC stain distinguish HNRNPC C1 from C2 or modified protein?
Do not assign separate C1 and C2 scores from this stain without an epitope map and isoform-specific validation (standard IHC interpretation). UniProt lists 4 isoforms, including C1 and C2, and describes a tetramer containing 3 C1 copies and 1 C2 copy (UniProt P07910 isoforms and subunit). It also lists an RRM at residues 16–87 and several phosphorylation sites, but the supplied antibody caption does not identify its epitope (UniProt P07910 domains and modified residues; datasheet A02726-1). Compare a second validated epitope or an orthogonal assay if isoform or modification status is central to the experiment (standard IHC practice).
How should I design an IF multiplex check for the nuclear IHC pattern?
Use IF as a separate localisation check and pair HNRNPC with a validated marker for the expected cell population, such as adipocytes in adipose tissue (HPA: High in adipocytes; standard IF practice). Select a spectrally separated, preferably far-red HNRNPC fluorophore after checking unstained tissue for autofluorescence (standard IF practice). Because the expected epitope is within the nucleus and HNRNPC has no transmembrane segment, optimise permeabilisation for nuclear antibody access rather than interpreting surface fluorescence as target signal (UniProt P07910 localisation and topology; standard IF practice). Include single-stain controls, a no-primary control and a nuclear counterstain, then compare nuclear overlap with the chromogenic pattern (standard IF practice; HPA subcellular).
How can I reduce diffuse brown staining without losing nuclear signal?
First inspect a no-primary section to separate secondary-reagent or chromogen background from primary-antibody staining (standard IHC practice). The documented assay used 10% goat serum blocking, a 1:100 primary dilution and an HRP secondary with DAB detection (datasheet A02726-1). Add a peroxidase-blocking step where appropriate, and optimise washes and DAB development using matched sections (standard chromogenic IHC practice). If background persists, titrate the primary around the documented dilution while keeping retrieval at EDTA pH 8.0 initially (datasheet A02726-1; standard IHC practice). Preserve nuclei as the reference compartment when judging improvement, since HNRNPC is nuclear (UniProt P07910 localisation).
What is a defensible way to quantify HNRNPC across IHC samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, then quantify nuclear staining rather than whole-section brown area (standard IHC practice; HPA tissue IHC). Report either the percentage of positive nuclei with an intensity-based H-score, or positive nuclei per mm² when cell density is the main outcome (standard IHC practice). Normalise positive nuclei to all assessable nuclei of the same cell population, or density to the measured viable tissue area (standard IHC practice). Keep retrieval, exposure to DAB and scoring thresholds consistent across compared sections, using the documented EDTA pH 8.0 condition as the starting protocol (standard IHC practice; datasheet A02726-1).
How do I distinguish genuine HNRNPC positivity from staining artefacts?
Treat staining in intact nuclei as the expected pattern, consistent with UniProt nuclear localisation and HPA’s general nuclear tissue profile (UniProt P07910 localisation; HPA tissue IHC). Confirm that positive nuclei belong to the cell population under study; HPA reports high staining in several cell types and low staining in soft-tissue fibroblasts (HPA tissue IHC). Discount isolated section-edge deposits, necrotic areas and diffuse cytoplasmic colour when they lack matching intact nuclear staining (standard IHC practice; UniProt P07910 localisation). Check a no-primary section and peroxidase blocking for endogenous enzyme or detection artefacts before calling unexpected brown signal positive (standard chromogenic IHC practice).
Boster reagents
Best HNRNPC / Heterogeneous nuclear ribonucleoproteins C1/C2 IHC Antibodies
IHC images cover human paraffin-embedded tissues (catalog IHC captions), and an IF image covers U2OS cells (A02726-1 IF caption). Both antibodies list Human, Mouse, and Rat reactivity (catalog reactivity).
Real IHC data
Anti-HNRNPC Antibody
Cat # A02726-1
A02726-1 has IHC images from human bladder urothelial carcinoma, breast cancer, testicular cancer, and esophageal squamous carcinoma, plus IF in U2OS cells (A02726-1 image captions). M02726 has a paraffin-embedded human colon IHC image and lists IF/ICC as applications (M02726 IHC caption; catalog applications).
Which to pick: For tissue IHC, choose A02726-1 for its illustrated paraffin-section conditions, including EDTA retrieval at pH 8.0; M02726 has an illustrated human colon paraffin section (each SKU’s IHC caption). For IF/ICC, choose A02726-1 if an IF image matters: it has one in U2OS cells, while M02726 lists IF/ICC without a supplied IF image (catalog IF captions; catalog applications). For cross-species work, both list Human, Mouse, and Rat reactivity; A02726-1 is polyclonal and M02726 is monoclonal, but their supplied IHC images show human tissue only and neither IHC caption reports the fixative (catalog reactivity; catalog clonality; catalog IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P07910 (HNRPC_HUMAN, Heterogeneous nuclear ribonucleoproteins C1/C2).
- Human Protein Atlas. HNRNPC tissue IHC expression (reliability: Supported).
- Human Protein Atlas. HNRNPC subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. HNRNPC antibody validation summary (5 antibodies).
- Elevated Heterogeneous Nuclear Ribonucleoprotein C Expression Correlates With Poor Prognosis in Patients With Surgically Resected Lung Adenocarcinoma. Frontiers in oncology 2020 — PMC7868529.
- M6A RNA Methylation Regulator HNRNPC Contributes to Tumorigenesis and Predicts Prognosis in Glioblastoma Multiforme. Frontiers in oncology 2020 — PMC7578363.
- HNRNPC reprograms cancer metabolism to drive acquired chemoresistance in colorectal cancer by repressing of Nrf2/SLC7A11-dependent ferroptosis. Cell communication and signaling : CCS 2025 — PMC12802253.
- A systematic pan-cancer study demonstrates the oncogenic function of heterogeneous nuclear ribonucleoprotein C. Aging 2022 — PMC9004556.
- PubMed PMID:2557628 — UniProt-cited evidence.
- PubMed PMID:3110598 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HNRNPC (Heterogeneous nuclear ribonucleoproteins C1/C2). Expected localisation Nuclear staining observed (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02726-1). Validated IHC antibody: A02726-1 at 1:50 recommended; image 1:100 (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human bladder urothelial carcinoma tissue; fixative not specified (datasheet A02726-1); 2. Fixation: Image fixative and duration unreported (datasheet A02726-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A02726-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02726-1); 8. Primary antibody: Rabbit anti-HNRNPC, 1:50 recommended; image 1:100 (datasheet A02726-1); 9. Primary incubation: Overnight at 4 °C (datasheet A02726-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Fibroblasts may show low staining (HPA tissue IHC); Isoform / epitope: 4 isoforms; check epitope before isoform claims (UniProt); Regulation: Regulation not annotated (UniProt). Evidence: UniProt P07910, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm., Datasheet A02726-1, PMC7868529, PMC7578363. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HNRNPC/immunohistochemistry-ihc-guide.