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EDF1 / Endothelial differentiation-related factor 1 · IHC design guide
Design Immunohistochemistry for EDF1
Plan EDF1 staining in paraffin sections using its reported cytoplasmic and nuclear tissue pattern (HPA tissue IHC). The guide covers controls, fixation consistency, antibody dilution of 1:50–1:100 (datasheet), and interpretation of tissue staining.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
EDF1 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 | Cytoplasmic and nuclear staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic and nuclear staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+2 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); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07668) | |
| Caveat | Adipocytes were not detected despite reported adipose abundance (HPA tissue IHC; UniProt) | |
| Regulation | More abundant in heart and pancreas (UniProt) | |
| Isoform / epitope | 3 isoforms; isoform-specific epitope coverage is unknown (UniProt) |
Section 1
Recommended EDF1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with one published EDF1 IHC method (PMC11792593).
| Sample | Paraffin-embedded rat brain tissue; fixative not specified (datasheet A07668) |
| Fixation | Image fixative and duration unreported (datasheet A07668); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-EDF1, 1:50-1:100 (datasheet A07668) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | EDF1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic and nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min (page retrieval rule: nuclear antigen; UniProt O60869: nucleus).
Section 2
What Is the Expected EDF1 Staining Pattern?
EDF1 should appear in cytoplasm and nuclei across many tissues, including adrenal glandular cells and bone marrow hematopoietic cells with medium staining (HPA: tissue IHC). HPA rates its tissue profile Approved, with medium consistency between staining and RNA data (HPA: tissue IHC). EDF1 has no transmembrane segment, so a membrane-only pattern is unexpected (UniProt O60869: topology).
| Cytoplasmic and nuclear staining in adrenal glandular cells or bone marrow hematopoietic cells. | This matches the reported compartments and medium staining in those cell populations (HPA: tissue IHC). Judge intensity against adjacent cells and controls; a mixed cytoplasmic and nuclear pattern is consistent with EDF1 (HPA: tissue IHC; UniProt O60869: subcellular location). |
| Crisp membrane-only, extracellular, or exclusively punctate staining in otherwise interpretable cells. | These patterns do not match the reported cytoplasmic and nuclear tissue profile (HPA: tissue IHC). A membrane-only result also conflicts with EDF1's lack of a transmembrane segment (UniProt O60869: topology). Review morphology, primary-antibody controls and detection before calling the pattern EDF1. |
| Strong staining confined to adipocytes, parathyroid glandular cells, or skeletal myocytes. | HPA reports EDF1 as not detected in these cell populations (HPA: tissue IHC). Treat an isolated strong result as suspect for nonspecific binding or endogenous detection activity; compare an expected-positive tissue and the appropriate detection control before interpreting it as biological expression (standard IHC practice). |
| Diffuse color across nuclei, cytoplasm, stroma and empty areas, with little cell-level contrast. | The reported EDF1 pattern is cellular and cytoplasmic/nuclear (HPA: tissue IHC). Color outside recognizable cells may reflect nonspecific reagent binding or detection background (standard IHC practice). It cannot establish EDF1 distribution until background controls and morphology are satisfactory. |
| No staining in an interpretable adrenal gland or bone marrow section. | Medium staining is reported in adrenal glandular and bone marrow hematopoietic cells (HPA: tissue IHC). Check that those cells are present, then review antibody application, retrieval, detection and controls (standard IHC practice). A single blank section does not establish EDF1 absence. |
💡Expected EDF1 appearanceCall a positive result when identifiable glandular or hematopoietic cells show medium cytoplasmic and nuclear staining (HPA: tissue IHC); isolated membrane-only or extracellular color is suspect (HPA: tissue IHC; UniProt O60869: topology).
| Tissue and cell selection | HPA reports a broad cytoplasmic and nuclear profile with low RNA tissue specificity, yet lists adipocytes, parathyroid glandular cells and skeletal myocytes as not detected by IHC (HPA: tissue IHC). Score the named cell population rather than assigning one result to a whole organ. |
| Compartment and stimulus context | EDF1 is cytoplasmic and nuclear; UniProt also reports nuclear localization upon NR5A1 binding or treatment of cells with TPA or forskolin (UniProt O60869: subcellular location). Nuclear prominence alone therefore needs cell and experimental context. |
| Antibody evidence | The listed antibody, HPA035642, is Approved for IHC and Supported for ICC (HPA: antibodies). The tissue profile has medium consistency between staining and RNA data (HPA: tissue IHC); use the reported pattern as a reference, not proof that every stained cell is specific. |
| Protein forms and chemistry | UniProt lists three isoforms and acetylation, phosphorylation and methylation among EDF1 annotations (UniProt O60869: isoforms; modified residues). These facts do not identify the IHC antibody's epitope or establish a particular retrieval condition or fixation sensitivity. |
| Processing and topology | UniProt lists no signal peptide, propeptide, transmembrane segment or glycosylation sites (UniProt O60869: processing; topology; glycosylation). Interpret convincing staining within cells; these annotations provide no basis for predicting a secreted or membrane-restricted IHC pattern. |
| IF/ICC Q: What should an EDF1-positive cell look like? | A: Mainly cytosolic signal, with additional nucleoplasmic and nucleolar signal, is reported in ICC-IF (HPA: subcellular). This cell-image finding complements the cytoplasmic and nuclear tissue IHC profile (HPA: tissue IHC); it does not specify an IHC protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected-positive cells have no chromogenic signal. | The positive cell population may be absent, or an assay step may have failed; adrenal glandular and bone marrow hematopoietic cells are reported at medium intensity (HPA: tissue IHC). | Verify cell identity and section integrity, then check the IHC antibody application, detection reagents and a concurrent positive control (standard IHC practice). Review retrieval against the antibody's validated IHC conditions; no EDF1-specific retrieval effect is supplied. |
| Every tissue compartment develops similar color. | Uniform color obscures the cell-level cytoplasmic and nuclear pattern (HPA: tissue IHC) and can arise from nonspecific binding or detection background (standard IHC practice). | Inspect a no-primary control, blocking and wash steps, and detection reagent exposure (standard IHC practice). Reassess only after cell boundaries and the expected compartments can be distinguished. |
| Color appears in the no-primary control. | The primary antibody cannot explain signal in its absence; endogenous detection activity or reagent background is possible (standard IHC practice). | Review the detection system's endogenous-activity control, blocking and chromogen development (standard IHC practice). Do not score tissue color as EDF1 while that control remains positive. |
| Staining is restricted to cell borders or extracellular material. | This differs from the HPA cytoplasmic and nuclear tissue profile and is unsupported by EDF1 topology (HPA: tissue IHC; UniProt O60869: topology). | Compare the pattern with an expected-positive cell population, inspect morphology and review primary-antibody and detection controls (standard IHC practice). Treat persistent border-only staining as unconfirmed. |
| An HPA not-detected cell population stains strongly. | HPA lists adipocytes, parathyroid glandular cells and skeletal myocytes as not detected, while its tissue evidence has medium consistency with RNA data (HPA: tissue IHC). | Confirm the stained cell type, compare an expected-positive section and inspect no-primary or other appropriate controls (standard IHC practice). Record any disagreement with HPA as unresolved rather than assigning specificity from color alone. |
| Nuclear staining dominates, with little visible cytoplasmic signal. | EDF1 can occupy both compartments, and UniProt reports nuclear localization in specific binding or treatment contexts (UniProt O60869: subcellular location); tissue IHC usually shows both (HPA: tissue IHC). | Assess matched cells and controls, then document the nuclear-to-cytoplasmic pattern and experimental context (standard IHC practice). Do not infer NR5A1 binding or TPA/forskolin exposure from staining alone. |
Experiment evidence
IHC & IF images showing EDF1 staining
A07668 · IHCImmunohistochemistry of paraffin-embedded rat brain using EDF1 antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10 mM
A07668 · IHCImmunohistochemistry of paraffin-embedded human gastric cancer using EDF1 antibody at dilution of 1:100 .Perform microwave antigen retrieval
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000107223-EDF1/tissue/cerebral+cortex#imid_8457420
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000107223-EDF1/tissue/colon#imid_8457362
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000107223-EDF1/tissue/liver#imid_8457363
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000107223-EDF1/tissue/kidney#imid_8457348
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035642; green = EDF1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035642; green = EDF1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for EDF1 IHC & IF
🧪Run appendix first: its glandular cells show Medium EDF1 staining (HPA: Appendix glandular cells, Medium). Use parathyroid gland as the negative tissue (HPA: Parathyroid glandular cells, Not detected); any independently verified EDF1-negative cells on the appendix slide should show counterstain without specific chromogen.
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show EDF1 in A-431, U-251MG, U2OS, KOLF2.1J, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched isotype controls using the primary antibody’s host species and isotype, plus an EDF1 knockout sample as a biological negative (standard IHC practice). Block endogenous peroxidase and check inflammatory cells in the appendix for residual enzyme-associated chromogen (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence; the selected A07668 paraffin-section caption also leaves the fixative unreported (A07668 tissue-IHC caption). That caption documents microwave retrieval in 10 mM PBS, pH 7.2, with antibody at 1:100 for rat brain, but does not establish that retrieval is required for appendix (A07668 tissue-IHC caption). There is no supplied comparison showing frozen sections or IF to be easier; endogenous peroxidase in appendix inflammatory cells may complicate chromogenic scoring (standard IHC practice).
HPA tissue IHC evidence for EDF1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced EDF1 IHC Tips
EDF1 staining can be cytoplasmic and nuclear; evaluate each compartment and cell population separately (UniProt O60869; HPA tissue IHC).
Which retrieval conditions should I start with for EDF1 paraffin-section IHC?
Start with Tris-EDTA at pH 9.0, heated to 95–98 °C for 20 min (page antigen-retrieval specification). Cool sections consistently before blocking, and compare nuclear and cytoplasmic staining in the same cell populations (standard IHC practice; HPA tissue IHC). If staining is weak, test microwave retrieval in 10 mM PBS, pH 7.2 as a separate condition; this was used with catalog antibody A07668 on paraffin-embedded rat brain (A07668 tissue-IHC caption). Keep antibody dilution and detection conditions constant during the comparison so a retrieval change is interpretable (standard IHC practice).
How should I troubleshoot EDF1 staining across differently fixed paraffin blocks?
EDF1-specific sensitivity to fixation is unknown from the supplied evidence; the A07668 rat-brain caption identifies paraffin embedding but does not state a fixative (A07668 tissue-IHC caption). Record each block’s fixative and fixation duration, then compare sections processed with the same retrieval, antibody incubation and detection conditions (standard IHC practice). Examine nuclear and cytoplasmic staining separately because both compartments are expected for EDF1 (UniProt O60869; HPA tissue IHC). If one block stains poorly, test retrieval conditions on matched sections before attributing the difference to fixation (standard IHC practice). Avoid treating a weak block as biologically negative without a suitable staining control (standard IHC practice).
Should EDF1 signal be nuclear, cytoplasmic, or both in tissue sections?
Score nuclear and cytoplasmic EDF1 separately: tissue IHC reports both in most tissues, while subcellular imaging supports a mainly cytosolic location with additional nucleoplasmic and nucleolar signal (HPA tissue IHC; HPA subcellular). UniProt also lists cytoplasmic and nuclear localisation, including nuclear localisation upon NR5A1 binding and TPA or forskolin treatment (UniProt O60869). Compare signal with cell morphology and a nuclear counterstain, and keep imaging and scoring thresholds consistent across sections (standard IHC practice). A change in nuclear-to-cytoplasmic distribution may be biologically plausible, but compartment shifts alone do not establish altered EDF1 abundance or activity (UniProt O60869; standard IHC interpretation).
Can this IHC result distinguish EDF1 isoforms or modified epitopes?
The supplied record lists 3 EDF1 isoforms, but provides no A07668 epitope map or isoform-specific IHC validation (UniProt O60869; A07668 tissue-IHC caption). Treat chromogenic signal as EDF1-associated staining without assigning it to isoform 1, 2 or 3 (UniProt O60869; standard IHC interpretation). EDF1 has reported acetylation, phosphorylation and methylation sites, yet the supplied evidence does not show how these modifications affect antibody binding (UniProt O60869). If isoform or modification specificity matters, obtain epitope and validation data and compare appropriate independently validated reagents on matched sections (standard IHC practice). Maintain identical retrieval and detection settings when evaluating any staining differences (standard IHC practice).
How can I use IF to examine EDF1 alongside a cell-identity marker?
For a complementary IF experiment, multiplex EDF1 with a validated marker for the cell population being examined, then assess colocalisation within individual cells (standard IF practice). EDF1 is reported in cytoplasm and nucleus and has no transmembrane segment, so use permeabilisation appropriate for intracellular epitopes and check that nuclear access is adequate (UniProt O60869; standard IF practice). Choose fluorophores whose emission is distinguishable from the specimen’s autofluorescence, and inspect single-marker controls before interpreting merged images (standard IF practice). Compare cytosol, nucleoplasm and nucleoli as distinct compartments where image resolution permits (HPA subcellular). The A07668 paraffin-section IHC caption does not establish an IF/ICC protocol for that antibody (A07668 tissue-IHC caption).
What should I check when EDF1 chromogenic staining looks diffuse or widespread?
EDF1 is reported in both cytoplasm and nucleus across most tissues, so widespread staining requires assessment by cell and compartment before it is called background (HPA tissue IHC). Inspect a no-primary control for nonspecific detection and ensure the general chromogenic workflow includes an appropriate peroxidase block before DAB development (standard IHC practice). Compare tissue interiors with section edges, folds and necrotic areas, where staining artefacts can complicate interpretation (standard IHC practice). If background persists, titrate the antibody and review blocking, wash stringency and DAB development time while holding retrieval constant (standard IHC practice). Document exposure and scoring thresholds consistently across controls and test sections (standard IHC practice).
How should I quantify EDF1 staining when nuclear and cytoplasmic patterns differ? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue area before scoring, then report nuclear and cytoplasmic EDF1 separately because both compartments are documented (HPA tissue IHC; UniProt O60869). An H-score can summarize the percentage of cells at each intensity; report percentage positive as well when intensity categories are uncertain (standard IHC practice). For spatial comparisons, count positive cells per mm² of evaluable tissue and normalize to the corresponding total cell count or evaluable area (standard IHC practice). Use the same counterstain, image settings and positivity thresholds across samples, and exclude folds and necrosis by a prespecified rule (standard IHC practice).
How do I separate credible EDF1 positivity from staining artefacts?
Credible EDF1 staining may be cytoplasmic or nuclear, with cytosol predominating in subcellular imaging; evaluate whether signal follows intact cells and their compartments (HPA tissue IHC; HPA subcellular). Check the expected cell population against the specimen: HPA reports medium staining in cerebral-cortex glial cells and no detected staining in skeletal-muscle myocytes (HPA tissue IHC). Strong signal confined to edges, folds or necrotic regions warrants an artefact check rather than immediate biological interpretation (standard IHC practice). Use a no-primary control and adequate peroxidase blocking to assess detection background, including endogenous enzyme activity during DAB development (standard IHC practice). Interpret weak or absent staining cautiously because HPA tissue IHC has approved reliability with medium consistency against RNA expression (HPA tissue IHC).
Boster reagents
Best EDF1 / Endothelial differentiation-related factor 1 IHC Antibodies
A07668 has paraffin-section IHC images from rat brain and human gastric cancer (catalog IHC image alts). IF/ICC is listed, but no IF image is supplied (catalog applications; catalog IF image alts).
Real IHC data
Anti-EDF1 Antibody
Cat # A07668
A07668 is the sole card and lists IHC, IF and ICC applications with Human, Mouse and Rat reactivity (catalog applications; catalog reactivity). Its IHC images show paraffin-embedded rat brain and human gastric cancer (catalog IHC image alts).
Which to pick: Choose A07668 for tissue IHC in paraffin sections, supported by its rat brain and human gastric cancer images; the fixative is unreported (catalog IHC image alts). Choose A07668 for IF/ICC because both applications are listed, while no IF image is supplied (catalog applications; catalog IF image alts). For cross-species work, A07668 lists Human, Mouse and Rat reactivity, although the supplied IHC images cover only human and rat samples (catalog reactivity; catalog IHC image alts).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O60869 (EDF1_HUMAN, Endothelial differentiation-related factor 1).
- Human Protein Atlas. EDF1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. EDF1 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the nucleoplasm and nucleoli..
- Human Protein Atlas. EDF1 antibody validation summary (1 antibodies).
- EDF1 accelerates ganglioside GD3 accumulation to boost CD52-mediated CD8(+) T cell dysfunction in neuroblastoma. Journal of experimental & clinical cancer research : CR 2025 — PMC11792593.
- PubMed PMID:9813014 — UniProt-cited evidence.
- PubMed PMID:10567391 — UniProt-cited evidence.
- PubMed PMID:15164053 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for EDF1 (Endothelial differentiation-related factor 1). Expected localisation Cytoplasmic and nuclear staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A07668 at 1:50-1:100 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat brain tissue; fixative not specified (datasheet A07668); 2. Fixation: Image fixative and duration unreported (datasheet A07668); 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-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-EDF1, 1:50-1:100 (datasheet A07668); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07668); Caveat: Adipocytes were not detected despite reported adipose abundance (HPA tissue IHC; UniProt); Isoform / epitope: 3 isoforms; isoform-specific epitope coverage is unknown (UniProt); Regulation: More abundant in heart and pancreas (UniProt). Evidence: UniProt O60869, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A07668, PMC11792593. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/EDF1/immunohistochemistry-ihc-guide.