HDGF / Hepatoma-derived growth factor · IHC design guide

Design Immunohistochemistry for HDGF

Plan HDGF paraffin IHC around distinct nuclear staining across tissues (HPA tissue IHC). This guide covers fixation, the catalog antibody’s 0.5–1 μg/mL IHC range (datasheet A01057), chromogenic detection, and nuclear scoring.

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 in tissues (HPA tissue IHC), antibody A01057, validated IHC image, and IHC protocol steps — Printable **HDGF** IHC protocol sheet — expected localisation Nuclear staining in 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 in tissues (HPA tissue IHC)
	Staining pattern	Distinct nuclear staining across tissues (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	Secretion may separate protein staining from tissue RNA (HPA tissue IHC)
	Regulation	No staining regulator specified (UniProt)
	Isoform / epitope	3 isoforms; epitope differences unspecified (UniProt)

Section 1

Recommended HDGF IHC & IF Protocols

The catalog antibody’s IHC protocol is followed by published HDGF staining protocols for synovial tissue, cervical adenocarcinoma, and endometrial carcinoma (PMC9169164; PMC4264238; 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 retrieval at pH 6 (datasheet A01057). The endometrial carcinoma study used 10 mM citrate at 100°C for 2 min (PMC3947826 methods).

Section 2

What Is the Expected HDGF Staining Pattern?

HDGF should show distinct nuclear staining across tissues in paraffin-section IHC (HPA tissue IHC: Enhanced; distinct nuclear expression in all tissues). High staining is reported in specified glandular, hematopoietic, respiratory epithelial, granular-layer, and endothelial cells (HPA tissue IHC: High in listed cells). Nucleoplasm is the main ICC-IF location (HPA subcellular: enhanced). HDGF also has cytoplasmic and secreted locations and no transmembrane segment (UniProt P51858).

What am I looking at on my slide?
Distinct nuclear signal in the expected cells, with interpretable counterstained nuclei.	This matches the reported IHC pattern (HPA tissue IHC: distinct nuclear expression in all tissues). High signal is documented in breast glandular cells and bone marrow hematopoietic cells, among other listed examples (HPA tissue IHC: High). Compare cells within the same section before assigning an intensity score.
Predominantly diffuse cytoplasmic signal, with little or no nuclear definition.	Question an IHC result that lacks the reported distinct nuclear pattern (HPA tissue IHC: nuclear expression). Cytoplasmic HDGF is biologically possible (UniProt P51858: cytoplasm), so compartment alone cannot prove artefact; assess whether the pattern persists with an independently validated antibody (HPA antibodies: 2 IHC Enhanced antibodies).
Strong staining in an unexpected cell population while the documented positive population is weak.	Consider cross-reactivity or endogenous chromogen-generating activity before calling that population HDGF positive (general IHC practice). For example, colon endothelial cells are documented as High (HPA tissue IHC: colon endothelial cells); staining elsewhere in colon needs its own validation. Broad tissue expression does not establish every cell type as High (UniProt P51858: ubiquitous; HPA tissue IHC: selected High cells).
Uniform haze, precipitate, or staining across tissue structures without clear nuclear contrast.	Treat this as background until controls and morphology support a cell-specific result (general IHC practice). Check blocking, washes, chromogen development, and a no-primary control (general IHC practice); the reported HDGF IHC pattern has distinct nuclei (HPA tissue IHC: nuclear expression).
No nuclear signal in a section expected to contain a documented High cell population.	This is an assay warning, not evidence that HDGF is absent (HPA tissue IHC: High in listed cells). First confirm that the relevant cells are present, then review antibody dilution, retrieval, detection, and control performance (general IHC practice). The supplied sources do not report HDGF-specific fixation sensitivity.

💡Expected HDGF appearanceCall a positive when expected cells show distinct, preferably strong nuclear staining with readable morphology (HPA tissue IHC: nuclear expression; High in listed cells); diffuse chromogen without nuclear definition is a suspect false positive (general IHC practice).

How each factor affects the staining
Antibody evidence	HPA048728 and HPA053422 are IHC Enhanced; CAB026035 is IHC Supported (HPA antibodies). An independently validated antibody can help resolve a disputed pattern (general IHC practice).
Antigen retrieval	Record and optimize the retrieval condition for the chosen IHC-validated antibody (general IHC practice). Neither HPA tissue IHC nor UniProt P51858 supplies an HDGF-specific retrieval or fixation effect.
Isoforms and epitope	UniProt lists 3 HDGF isoforms and a PWWP domain at residues 12–69 (UniProt P51858). The supplied evidence does not identify antibody epitopes or establish isoform-specific staining.
Intracellular versus secreted protein	Nuclear IHC is the documented tissue pattern (HPA tissue IHC). HDGF can also occur in cytoplasm and inside secreted exosomes, and may be released as free protein (UniProt P51858); extracellular staining alone needs corroboration.
IF/ICC: where should signal appear?	Primarily in the nucleoplasm (HPA subcellular: enhanced); HPA reports ICC Enhanced for HPA048728 and HPA053422 (HPA antibodies). This localisation cross-check does not define an IF/ICC protocol.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Known-positive cells have no nuclear signal.	The expected cells may be absent, or the IHC run may have failed (HPA tissue IHC: listed High cells; general IHC practice).	Confirm the cell population on the section; check a positive control, antibody dilution, retrieval, and detection in that order (general IHC practice). Do not infer HDGF-specific fixation loss from this result.
Nuclei stain weakly or unevenly across a positive section.	Section handling, retrieval, reagent coverage, or development may vary across the slide (general IHC practice).	Compare staining with tissue morphology and a control section; repeat with consistent section coverage and the antibody's validated IHC conditions (general IHC practice).
Diffuse chromogen obscures nuclear boundaries.	Background from insufficient washing, blocking, or excess antibody or chromogen is possible (general IHC practice).	Run a no-primary control, improve washing or blocking, and titrate antibody or development as needed (general IHC practice). Score only distinguishable cellular staining.
Signal is chiefly cytoplasmic or extracellular.	These locations are biologically plausible (UniProt P51858: cytoplasm; exosomal secretion), but differ from the distinct nuclear tissue-IHC profile (HPA tissue IHC).	Check nuclear counterstain and controls, then compare with an independent IHC Enhanced antibody if available (HPA antibodies: HPA048728, HPA053422; general IHC practice).
A different cell population stains more strongly than the documented High cells.	Cross-reactivity or endogenous detection activity is possible (general IHC practice); HPA's listed High cells do not validate every stained population (HPA tissue IHC).	Identify cells by morphology, inspect a no-primary control, and seek agreement with another validated antibody before assigning HDGF expression (general IHC practice).
IHC appears nuclear, but a companion IF/ICC image appears diffuse.	HPA reports enhanced nucleoplasmic ICC-IF localisation (HPA subcellular); diffuse fluorescence can arise from background or image settings (general IF practice).	Review IF controls, nuclear counterstain, exposure, and cell boundaries (general IF practice). Interpret each assay with its own controls; no IF/ICC protocol is specified here.

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 show HDGF staining (HPA: High in hematopoietic cells), with a predominantly nuclear pattern (HPA: nucleoplasm). HPA detects HDGF in all 45 scored tissues, so no negative tissue is supported; use no-primary and isotype controls, and expect anucleate erythrocytes on the positive slide to lack nuclear staining, while treating other unstained cells as a within-slide comparison rather than proven HDGF-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 slide; a concentration-matched rabbit IgG control matched to the primary antibody’s clonality; and HDGF-knockout material processed in parallel as a biological negative (caption: rabbit anti-HDGF A01057; standard IHC practice). Block endogenous peroxidase in bone marrow and check for residual chromogen signal in the no-primary slide (HPA: bone marrow hematopoietic cells; standard IHC practice).

⚠️Feasibility: A tissue IHC example reports citrate retrieval at pH 6 for 20 minutes in a paraffin section, but its fixative is unreported (A01057 tissue-IHC caption). No target-specific fixation window or fixation effect is reported in the supplied evidence, and the caption does not establish fixation; retrieval performance should therefore be checked under the chosen processing conditions (A01057 tissue-IHC caption). IHC-P has a direct tissue example, while IF/ICC has HPA cell images; the evidence does not establish that frozen sections or IF are easier (A01057 tissue-IHC caption; HPA: ICC-IF images). The selected A01057 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image A01057).

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

Use nuclear staining as the main reference for HDGF in chromogenic paraffin-section IHC, while evaluating cytoplasmic or extracellular signal separately (HPA tissue IHC; UniProt P51858).

What retrieval conditions should I try first for HDGF in paraffin sections?

Start with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A01057; caption A01057). The documented mouse-liver paraffin-section example used those conditions before overnight incubation with 1 μg/mL antibody at 4°C (caption A01057). Keep heating, cooling, section thickness, and detection conditions consistent when comparing retrieval runs (standard IHC practice). If nuclear staining remains weak, test a separately labeled alternative retrieval condition on adjacent sections while retaining citrate pH 6 as the reference (standard IHC practice). Judge improvement by nuclear signal and tissue preservation, since HDGF has distinct nuclear expression across tissues (HPA tissue IHC).

How should I troubleshoot HDGF staining when fixation conditions differ?

Target-specific sensitivity to fixative type or fixation duration is unknown from the supplied evidence (caption A01057: fixative not stated). Record the actual fixative, time in fixative, tissue thickness, processing schedule, and section age before comparing slides (standard IHC practice). The documented paraffin-section result cannot establish an optimal fixation condition because its caption gives no fixative (caption A01057). If staining differs between batches, compare similarly processed control sections using citrate pH 6 retrieval for 20 minutes and the same detection settings (caption A01057; standard IHC practice). Interpret any resulting fixation effect as an observation of that comparison, rather than a known HDGF-specific property (standard IHC practice).

Which compartments should show convincing HDGF staining?

Prioritize nuclear signal: tissue IHC shows distinct nuclear expression across tissues, and ICC/IF localizes HDGF to the nucleoplasm (HPA tissue IHC; HPA subcellular). Cytoplasmic signal is biologically possible, and HDGF can occur inside extracellular exosomes or be released as free protein (UniProt P51858). Evaluate those patterns separately from nuclear staining rather than counting every brown deposit as an equivalent positive cell (standard IHC practice). Check whether signal follows intact cell boundaries and persists away from cut edges, folds, and necrotic areas (standard IHC practice). A predominantly extracellular pattern needs additional controls before assignment to HDGF, because the strongest tissue-IHC reference pattern is nuclear (HPA tissue IHC; standard IHC practice).

Could epitope placement or HDGF isoforms explain discordant staining?

HDGF has 3 annotated isoforms and a PWWP domain at residues 12–69 (UniProt P51858). It also has reported modifications, including acetylation at residue 44 and phosphorylation at residues 132, 133, 165, and 184 (UniProt P51858). The supplied caption does not identify the A01057 binding epitope, so it cannot establish which isoforms or modified forms the staining detects (caption A01057). When two antibodies disagree, compare their documented immunogens or epitopes, then assess staining on adjacent sections under matched retrieval and detection conditions (standard IHC practice). Avoid interpreting a compartment difference as isoform-specific without independent isoform-sensitive evidence (standard IHC practice).

How can IF help check the cell identity and location seen by IHC?

Use a separate IF experiment to compare HDGF with a marker for the expected cell type; for example, evaluate glandular cells in breast, where tissue IHC reports high HDGF staining (HPA tissue IHC; standard IF practice). Include a nuclear counterstain to assess the nucleoplasmic pattern reported by ICC/IF (HPA subcellular; standard IF practice). Choose spectrally separated fluorophores and inspect unstained tissue so autofluorescence is not mistaken for HDGF (standard IF practice). For intracellular nuclear epitopes, optimize permeabilisation to give antibody access through cell and nuclear membranes while preserving morphology (UniProt P51858 topology; standard IF practice). Treat IF colocalization as supporting evidence and score the chromogenic paraffin sections independently (standard IHC/IF practice).

What should I check when HDGF DAB staining looks diffuse?

The documented paraffin-section procedure used 10% goat-serum blocking, biotinylated secondary antibody, an avidin–biotin detection complex, and DAB (caption A01057). For diffuse staining, compare a no-primary control and inspect endogenous peroxidase and biotin contributions under the same detection conditions (standard IHC practice). Apply a peroxidase block as a general chromogenic IHC step, and assess whether blocking, washing, or primary-antibody concentration improves the nuclear-to-background ratio (standard IHC practice). The caption's primary concentration was 1 μg/mL with overnight incubation at 4°C (caption A01057). Review extracellular deposits cautiously because HDGF may be secreted, while the tissue reference pattern is distinctly nuclear (UniProt P51858; HPA tissue IHC).

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

Define the scoring compartment before analysis; nuclear staining has the clearest tissue-IHC reference pattern for HDGF (HPA tissue IHC). Within annotated regions, record the percentage of positive cells and nuclear intensity, or calculate an H-score from intensity categories applied consistently across slides (standard IHC practice). For selected cell populations, report positive cells per mm² alongside the total count of eligible cells or tissue area analyzed (standard IHC practice). Normalize comparisons to the same cell type, viable tissue region, exposure to detection reagents, and scoring threshold (standard IHC practice). Report cytoplasmic or extracellular staining separately because HDGF also has those annotated locations (UniProt P51858).

How do I distinguish genuine HDGF staining from section artefacts?

Convincing HDGF IHC should include a reproducible nuclear pattern in intact cells, consistent with the distinct nuclear expression reported across tissues (HPA tissue IHC; standard IHC practice). Compare the stained cells with tissue architecture; high signal is reported in several specific populations, including glandular cells in breast and endothelial cells in colon (HPA tissue IHC). Treat staining confined to cut edges, folds, necrosis, or disrupted regions as suspect until it persists in intact areas (standard IHC practice). Check a no-primary section and the peroxidase-blocked workflow when brown deposits could reflect endogenous enzyme activity (standard IHC practice). Assess cytoplasmic or extracellular signal independently because those locations are biologically plausible for HDGF (UniProt P51858).

Boster reagents

Best HDGF / Hepatoma-derived growth factor IHC Antibodies

A01057 has IHC images from human, mouse and rat tissues and an IF image from U20S cells; M01057-1 is listed for human and mouse IHC/IF (catalog: image captions, applications, reactivity).

Real IHC data

Anti-HDGF Antibody ®

Cat # A01057

View product →

A01057 has paraffin-section IHC images from mouse and rat liver and human intestinal and lung cancer tissues, plus an IF image from U20S cells (catalog: image captions). M01057-1 is listed for human and mouse IHC and IF/ICC, but has no supplied IHC or IF image (catalog: applications, reactivity, image captions).

Which to pick: For tissue IHC, choose A01057 when a matching paraffin-section example matters; its caption documents citrate retrieval at pH 6 for 20 minutes and primary antibody at 1 μg/ml overnight at 4°C (A01057: IHC captions). For IF/ICC, A01057 has a U20S cell IF example at 2 μg/ml, while M01057-1 offers a rabbit monoclonal option, clone 26H94, listed for IF/ICC without a supplied image (A01057: IF caption; M01057-1: catalog applications and clone). For cross-species tissue work, A01057 covers human, mouse and rat; M01057-1 lists human and mouse, with no supplied IHC processing example, and the fixative is unreported in A01057’s paraffin-section captions (catalog: reactivity and IHC captions).

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.