HDGF / Hepatoma-derived growth factor · IHC design guide

Design Immunohistochemistry for HDGF

HDGF paraffin IHC shows distinct nuclear staining across tissues (HPA tissue IHC). Start with the IHC-validated antibody at 0.5–1 μg/mL (datasheet A01057), and score nuclear staining consistently across sections (standard IHC practice).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for HDGF (IHC for HDGF): expected localisation Nuclear staining across tissues (HPA tissue IHC), antibody A01057, validated IHC image, and IHC protocol steps — Printable **HDGF** IHC protocol sheet — expected localisation Nuclear staining across tissues (HPA tissue IHC), antibody A01057, controls and protocol steps. Open the full HDGF IHC guide →

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HDGF 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 across tissues (HPA tissue IHC)
	Staining pattern	Distinct nuclear staining in tissue cells (HPA tissue IHC)
	Antigen retrieval	Citrate pH 6 HIER, heat-mediated (datasheet A01057)
	Positive control ⓘ	Adrenal gland+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	Exosomal secretion may complicate tissue interpretation (UniProt)
	Regulation	Ubiquitous expression (UniProt)
	Isoform / epitope	3 isoforms; check epitope coverage (UniProt)

Section 1

Recommended HDGF IHC & IF Protocols

Compare the catalog antibody’s IHC-P protocol (datasheet A01057) with published HDGF staining protocols for synovial tissue (PMC9169164) and endometrial carcinoma (PMC3947826).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded mouse liver tissues; fixative not specified (datasheet A01057)
Fixation	Image fixative and duration unreported (datasheet A01057); 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 A01057)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A01057)
Primary antibody	Rabbit anti-HDGF, 0.5-1μg/ml (datasheet A01057)
Primary incubation	Overnight at 4 °C (datasheet A01057)
Detection	Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01057)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	HDGF-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Distinct nuclear expression in all tissues. No signal in the no-primary control.

💡Decision noteStart with heat-mediated citrate pH 6 retrieval for the catalog antibody (datasheet A01057); the endometrial study used 10 mM citrate at 100°C for 2 min (PMC3947826).

Section 2

What Is the Expected HDGF Staining Pattern?

HDGF should show distinct nuclear staining across tissues (HPA: Enhanced tissue IHC; distinct nuclear expression in all tissues), including glandular, hematopoietic, respiratory epithelial, endothelial, and cerebellar granular-layer cells reported as high (HPA: tissue IHC). UniProt also places HDGF in the cytoplasm and inside secreted exosomes; it has no transmembrane segment (UniProt P51858).

What am I looking at on my slide?
Distinct nuclear staining in glandular cells, such as adrenal gland, appendix, breast, or cervix (HPA: High in these cells).	This matches the dominant tissue pattern (HPA: distinct nuclear expression in all tissues). Compare nuclei within the tissue and score the specified cell population; staining elsewhere alone does not establish the expected pattern.
Predominantly membrane-like staining, with little or no nuclear signal in an expected positive cell population.	This conflicts with the observed tissue pattern (HPA: distinct nuclear expression) and lacks support from HDGF topology (UniProt P51858: no transmembrane segment). Check section morphology, detection background, and antibody specificity. Cytoplasmic signal alone is less decisive because UniProt also lists cytoplasm (UniProt P51858).
Strong signal in a cell population other than the one being assessed, while its expected nuclei are unstained.	Consider cross-reactivity or endogenous detection activity; verify the cell identity and review controls (general IHC practice). Do not label that population intrinsically HDGF-negative: UniProt calls expression ubiquitous, and the supplied HPA profile lists no negative tissue (UniProt P51858; HPA: tissue IHC).
Diffuse color covers nuclei, cytoplasm, and tissue spaces without crisp cellular boundaries.	This cannot be scored as the distinct nuclear pattern (HPA: tissue IHC). Review blocking, washing, antibody concentration, and chromogen development with appropriate controls (general IHC practice); the pattern alone cannot identify which step caused the background.
No detectable nuclear signal in a supplied high-staining reference, such as bone-marrow hematopoietic cells (HPA: High).	Treat the run as potentially insensitive before interpreting another sample as negative. Check tissue integrity, retrieval, antibody dilution, and detection reagents (general IHC practice). The supplied record does not establish a negative tissue reference or HDGF-specific retrieval conditions.

💡Expected HDGF appearanceCall the slide positive when a listed cell population shows distinct, readily discernible nuclear signal (HPA: Enhanced tissue IHC; High in listed cells); diffuse color or membrane-only staining is a warning pattern (HPA: nuclear tissue profile; UniProt P51858 topology).

How each factor affects the staining
Antibody validation	HPA048728 and HPA053422 have Enhanced IHC status; CAB026035 is Supported (HPA: antibody validation). Enhanced supports the reported pattern through independent antibodies or orthogonal data; it does not make every stain in a new sample specific.
Retrieval and assay settings	Optimize retrieval and dilution against a listed high-staining tissue and appropriate controls (general IHC practice; HPA: High in listed populations). No HDGF-specific retrieval condition, dilution, or fixation sensitivity is supplied; avoid attributing a failed stain to a particular retrieval or fixation effect.
Isoforms and epitope coverage	UniProt lists three HDGF isoforms, a PWWP domain at residues 12–69, and several modified residues (UniProt P51858). The supplied sources give no catalog-antibody epitope, so they cannot predict isoform coverage or a modification-dependent staining change.
Secreted HDGF and compartment calls	HDGF can occur inside secreted exosomes, and free-protein secretion is proposed (UniProt P51858). HPA nevertheless reports distinct nuclear tissue staining (HPA: tissue IHC). An extracellular-looking deposit alone is insufficient to confirm specific HDGF staining.
IF/ICC Q: What localisation should be expected?	A: Nucleoplasmic signal is the principal ICC-IF observation (HPA: Nucleoplasm, enhanced). UniProt additionally lists cytoplasm (UniProt P51858). Use the separate IF/ICC guide for its workflow; the IHC tissue pattern here does not supply an IF protocol.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Known high-staining tissue gives no nuclear signal (HPA: High in listed populations).	Insufficient retrieval, overly dilute primary antibody, or failed detection are possible (general IHC practice); the pattern does not distinguish them.	Confirm the expected cell population, then check retrieval and antibody dilution against controls and verify that detection reagents work (general IHC practice).
Color appears throughout the section rather than in defined nuclei.	Incomplete blocking or washing, concentrated antibody, or prolonged chromogen development may contribute (general IHC practice).	Review a no-primary control; adjust blocking, washing, dilution, or development one variable at a time (general IHC practice).
Unexpected cells stain strongly, but expected nuclei do not.	Cross-reactivity or endogenous detection activity is possible (general IHC practice); HDGF's reported ubiquitous expression prevents a simple negative-cell assumption (UniProt P51858).	Identify the cells morphologically, check no-primary and detection controls, and compare the nuclear pattern with a listed high-staining population (general IHC practice; HPA: tissue IHC).
The signal appears mainly at cell borders.	A border-only pattern differs from HPA's nuclear tissue pattern (HPA: tissue IHC) and is unsupported by HDGF topology (UniProt P51858: no transmembrane segment).	Check morphology and background controls; seek reproducible nuclear staining before assigning the border signal to HDGF (general IHC practice; HPA: tissue IHC).
A sample has weak nuclear staining relative to a listed high-staining reference.	Sample variation or reduced assay sensitivity may explain the difference; HPA's high calls describe the listed cells, not every specimen (HPA: tissue IHC).	Compare like cell populations and inspect the reference and controls in the same run before scoring intensity (general IHC practice).
Extracellular-looking staining is the only positive feature.	Exosomal HDGF is biologically plausible (UniProt P51858), but extracellular color alone does not establish its identity or reproduce HPA's nuclear tissue profile (HPA: tissue IHC).	Assess expected nuclei and background controls first; report the extracellular pattern separately without calling it a confirmed HDGF localisation (general IHC practice).

Source: UniProt P51858 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing HDGF staining

IHC analysis of HDGF using anti-HDGF antibody (A01057). HDGF was detected in paraffin-embedded section of mouse liver tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-HDGF Antibody (A01057) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

A01057 · IHCIHC analysis of HDGF using anti-HDGF antibody (A01057). HDGF was detected in paraffin-embedded section of mouse liver tissues. Heat mediated

IHC analysis of HDGF using anti-HDGF antibody (A01057). HDGF was detected in paraffin-embedded section of rat liver tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-HDGF Antibody (A01057) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

A01057 · IHCIHC analysis of HDGF using anti-HDGF antibody (A01057). HDGF was detected in paraffin-embedded section of rat liver tissues. Heat mediated a

IF analysis of HDGF using anti-HDGF antibody (A01057). HDGF was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-HDGF Antibody (A01057) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

A01057 · IFIF analysis of HDGF using anti-HDGF antibody (A01057). HDGF was detected in immunocytochemical section of U20S cells. Enzyme antigen retriev

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000143321-HDGF/tissue/cerebral+cortex#imid_17089707

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000143321-HDGF/tissue/colon#imid_17089633

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000143321-HDGF/tissue/liver#imid_17089642

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000143321-HDGF/tissue/kidney#imid_17089635

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB026035; green = HDGF, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for HDGF IHC & IF

🧪Run bone marrow first: hematopoietic cells should stain strongly (HPA: High in hematopoietic cells). HPA detects HDGF in all 45 scored tissues, so use no-primary and isotype controls for the negative; other cells on the positive slide should show minimal nonspecific background, but should not be treated as target-negative cells (HPA: no negative tissue rows; standard IHC practice).

Positive control tissue: Adrenal gland (Glandular cells, HPA High)

Negative control tissue: None in HPA: HDGF 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 HDGF in A-431, U-251MG, U2OS, MCF-7, NIH 3T3, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only control, matched rabbit IgG isotype control, and HDGF-knockout tissue as a biological negative (caption: rabbit primary; standard IHC practice). Block endogenous peroxidase and assess residual background in bone marrow before interpreting DAB staining (HPA: bone marrow positive; standard IHC practice).

⚠️Feasibility: The selected A01057 paraffin-section caption used citrate retrieval at pH 6 for 20 minutes, but does not establish that retrieval is required; its fixative is unreported (caption: citrate retrieval; fixative not stated). No target-specific fixation window or fixation effect is reported, and the supplied evidence does not establish whether frozen sections or IF are easier (supplied target/application evidence). Bone marrow may show endogenous peroxidase signal, so interpret chromogenic staining against the peroxidase-blocked controls (HPA: bone marrow positive; standard IHC practice).

HPA tissue IHC evidence for HDGF

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex.). 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
None in HPA: HDGF is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.

Section 3

Advanced HDGF IHC Tips

Troubleshoot chromogenic HDGF staining in paraffin sections using the catalog antibody’s tissue example, localisation evidence, and matched controls.

How should I retrieve HDGF antigen when nuclear staining is weak?

Start with heat mediated citrate retrieval at pH 6 for 20 minutes (datasheet A01057). The catalog antibody’s paraffin section example used this treatment before overnight incubation at 1 μg/mL and 4°C (datasheet A01057). If nuclei remain weak, adjust heating time in small increments while holding antibody concentration and detection conditions constant; compare serial sections with an established positive control (general IHC practice). Record section loss and background alongside signal, since excessive heating can damage morphology and complicate nuclear scoring (general IHC practice). The example identifies paraffin embedding but does not report a fixative, so retrieval performance under a particular fixation regimen is unestablished (datasheet A01057).

Could fixation explain inconsistent HDGF staining across paraffin sections?

Target specific sensitivity of HDGF staining to fixation is unknown from the supplied evidence; the catalog antibody’s paraffin section caption does not state its fixative (datasheet A01057). Record the actual fixative, fixation duration, tissue thickness, processing schedule, and block age for each specimen before attributing differences to biology (general IHC practice). Compare similarly processed serial sections under the same citrate pH 6 retrieval and antibody incubation conditions used in the example (datasheet A01057). Check preserved nuclear detail and tissue integrity while adjusting processing variables one at a time (general IHC practice). Nuclear staining patterns and HDGF modification sites do not establish a fixation effect (HPA tissue IHC; UniProt P51858).

What localisation should I expect when assessing HDGF IHC?

Expect a prominent nuclear signal: tissue IHC reports distinct nuclear expression across tissues, and cell imaging places HDGF in the nucleoplasm (HPA tissue IHC; HPA subcellular). HDGF is also annotated in the cytoplasm and within secreted extracellular exosomes, with no transmembrane segment (UniProt P51858). Score nuclear and cytoplasmic compartments separately rather than treating all brown signal as equivalent, and retain cell type information for each region (general IHC practice; UniProt P51858 localisation). If an extracellular pattern appears, test whether it survives matched detection controls before interpreting it as secretion (general IHC practice; UniProt P51858 localisation). HPA’s tissue profile supports broad nuclear expression, not equal staining intensity in every specimen (HPA tissue IHC).

How can epitope choice affect interpretation of HDGF staining?

HDGF has 3 annotated isoforms and a PWWP domain spanning residues 12–69, but the supplied catalog example does not identify the antibody epitope (UniProt P51858; datasheet A01057). Therefore, staining cannot establish which isoform is present or whether a negative region lacks every HDGF variant (UniProt P51858 isoforms; datasheet A01057). Document the immunogen and epitope range if available, then compare variant coverage before interpreting discordant results (general IHC practice). Annotated modifications include acetylation at residue 44 and phosphorylation at residues 132, 133, 165, and 184 (UniProt P51858). Their effect on this antibody’s binding is untested in the supplied evidence (datasheet A01057).

How should I check an HDGF pattern by multiplex immunofluorescence?

Use the separate IF/ICC workflow to assess nuclear signal alongside a marker identifying the cell population of interest; HDGF is reported in the nucleoplasm (HPA subcellular). Choose fluorophores after inspecting unstained tissue channels for autofluorescence, and include single stain controls to detect spectral bleed through (general IF practice). For an intracellular nuclear epitope, permeabilise cells or sections sufficiently for antibody access; HDGF has no transmembrane segment, while exosome associated HDGF is reported inside exosomes (UniProt P51858 topology and localisation; general IF practice). Assess nuclear and cytoplasmic channels independently, then compare the cell pattern with chromogenic IHC without assuming identical epitope access (HPA tissue IHC; general IF practice). The paraffin IHC caption supplies no IF fixation or dilution conditions (datasheet A01057).

What should I change when DAB background obscures HDGF nuclei?

Check a section without primary antibody and inspect whether brown signal tracks tissue edges, damaged areas, or endogenous enzyme activity (general IHC practice). The catalog example used 10% goat serum, a biotinylated secondary antibody, a streptavidin biotin complex, and DAB (datasheet A01057). Apply an appropriate peroxidase block before DAB development, and evaluate biotin related background with a matched detection control when using this detection system (general IHC practice; datasheet A01057). If diffuse staining persists, titrate the primary around the example’s 1 μg/mL while keeping retrieval and development time fixed (datasheet A01057; general IHC practice). Preserve the expected nuclear pattern as the main signal check (HPA tissue IHC).

How should I quantify HDGF staining across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and tissue region before scoring, since HDGF is broadly expressed and HPA reports distinct nuclear staining across tissues (UniProt P51858 tissue specificity; HPA tissue IHC). For chromogenic sections, report the percentage of positive nuclei and an intensity weighted H-score, or positive cell density per mm² when cell counts and area are reliable (general IHC practice). Normalise to the number of evaluable cells or the analysed viable tissue area, and use identical thresholds across batches (general IHC practice). Score cytoplasmic signal separately from nuclear signal because both localisations are annotated (UniProt P51858 localisation). Exclude folds, edges, necrosis, and failed counterstaining before comparing cases (general IHC practice).

How do I distinguish genuine HDGF positivity from staining artefact?

A reproducible nuclear pattern in intact cells supports the expected result, because tissue IHC reports distinct nuclear expression and cell imaging places HDGF in the nucleoplasm (HPA tissue IHC; HPA subcellular). Check whether staining follows the relevant cells rather than an edge, fold, or necrotic zone, and compare a section lacking primary antibody (general IHC practice). Cytoplasmic signal is biologically plausible, while HDGF has no transmembrane segment; a membrane only rim warrants additional scrutiny (UniProt P51858 localisation and topology). Diffuse brown deposits that remain in the detection control suggest endogenous enzyme activity or other detection background (general IHC practice). Interpret extracellular signal cautiously because HDGF can be secreted within exosomes (UniProt P51858 localisation).

Boster reagents

Best HDGF / Hepatoma-derived growth factor IHC Antibodies

A01057 has real IHC images from human, mouse and rat paraffin sections and IF data from U20S cells (catalog image captions); M01057-1 lists human/mouse IHC and IF applications (catalog applications/reactivity).

Real IHC data

Anti-HDGF Antibody ®

Cat # A01057

View product →

A01057 will render with a mouse liver IHC figure; its captions also document rat liver, human intestinal cancer and human lung cancer paraffin sections, plus U20S cell IF (A01057 image captions). M01057-1 will render as a rabbit monoclonal antibody, clone 26H94, listed for human/mouse IHC and ICC/IF, with no IHC or IF image caption supplied (M01057-1 catalog clone/applications/reactivity/image alts).

Which to pick: Choose A01057 for tissue IHC: its own captions show paraffin-section staining in human, mouse and rat samples at 1 μg/ml after citrate retrieval, while the fixative is unreported (A01057 IHC image captions). For IF/ICC, A01057 has a U20S cell IF image at 2 μg/ml; M01057-1 is a rabbit monoclonal option listed for ICC/IF without a supplied IF image (A01057 IF image caption; M01057-1 catalog clone/applications/image alts). For cross-species work, A01057 has documented human, mouse and rat IHC images, whereas M01057-1 lists human and mouse reactivity (A01057 IHC image captions; M01057-1 catalog reactivity).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry P51858 (HDGF_HUMAN, Hepatoma-derived growth factor).
Human Protein Atlas. HDGF tissue IHC expression (reliability: Enhanced).
Human Protein Atlas. HDGF subcellular location (ICC-IF): Localized to the nucleoplasm..
Human Protein Atlas. HDGF antibody validation summary (3 antibodies).
Association of serum hepatoma-derived growth factor levels with disease activity in rheumatoid arthritis: A pilot study. Journal of clinical laboratory analysis 2022 — PMC9169164.
Hepatoma-derived growth factor upregulation is correlated with prognostic factors of early-stage cervical adenocarcinoma. International journal of molecular sciences 2014 — PMC4264238.
The regulatory role of hepatoma-derived growth factor as an angiogenic factor in the eye. Molecular vision 2016 — PMC4842004.
High nuclear expression of HDGF correlates with disease progression and poor prognosis in human endometrial carcinoma. Disease markers 2014 — PMC3947826.
PubMed PMID:7929202 — UniProt-cited evidence.
PubMed PMID:14702039 — UniProt-cited evidence.
PubMed PMID:16710414 — UniProt-cited evidence.