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TOE1 / Target of EGR1 protein 1 · IHC design guide
Design Immunohistochemistry for TOE1
Plan TOE1 staining in paraffin sections with the IHC-validated antibody at 2–5 μg/ml (datasheet A09852-2). Compare the nuclear pattern with tissue IHC evidence, using cerebellar Purkinje cells as a high-staining reference (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TOE1 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 | General nuclear staining (HPA tissue IHC) | |
| Staining pattern | Nuclear staining in many cell types, including Purkinje cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A09852-2) | |
| 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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A09852-2) | |
| Caveat | Cholangiocytes may stain weakly (HPA tissue IHC) | |
| Regulation | Widely expressed; no regulator annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended TOE1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA pH 8.0 retrieval (datasheet A09852-2). Two published TOE1 IHC protocols offer additional conditions (PMC8092974; PMC12764802).
| Sample | Paraffin-embedded human cerebellum tissue; fixative not specified (datasheet A09852-2) |
| Fixation | Image fixative and duration unreported (datasheet A09852-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A09852-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A09852-2) |
| Primary antibody | Rabbit anti-TOE1, 2-5μg/ml (datasheet A09852-2) |
| Primary incubation | Overnight at 4 °C (datasheet A09852-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A09852-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TOE1-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 pH 8.0 for the catalog antibody (datasheet A09852-2); both published protocols used citrate retrieval (PMC8092974; PMC12764802).
Section 2
What Is the Expected TOE1 Staining Pattern?
TOE1 should appear chiefly in nuclei across many cell types (UniProt Q96GM8: nucleus, nucleolus and nuclear speckles; widely expressed; no transmembrane segment). HPA describes general nuclear tissue staining, including High staining in adipocytes, hematopoietic cells and several epithelial or neural cell populations (HPA tissue IHC: Supported). That assessment reflects agreement with RNA data and awaits external verification (HPA tissue IHC: reliability description).
| Nuclear chromogen in adipocytes, bone marrow hematopoietic cells or bronchial respiratory epithelial cells. | This matches the reported High cell-specific staining and general nuclear profile (HPA tissue IHC). Score staining in the named cells and their nuclei, rather than treating every cell in the section as an equally strong positive (HPA tissue IHC: cell-level observations). |
| Predominantly cytoplasmic, membranous or extracellular staining, with little nuclear signal. | That compartment pattern conflicts with TOE1's nuclear localization and lack of a transmembrane segment (UniProt Q96GM8). Check localization against the counterstain and a reported positive cell population before interpreting the deposit as TOE1 (HPA tissue IHC: general nuclear expression; standard IHC practice). |
| Strong signal in an unexpected cell population, especially where HPA reports Low staining. | HPA reports Low staining in parathyroid glandular cells and liver cholangiocytes; it supplies no negative tissue here (HPA tissue IHC). Strong staining there warrants checks for antibody cross-reactivity and endogenous detection activity (standard IHC practice). Low does not mean absent (HPA tissue IHC: Low). |
| Diffuse color across nuclei, cytoplasm and tissue spaces. | A widespread deposit that obscures cell boundaries prevents a reliable nuclear call (standard IHC practice). Assess background with an appropriate primary-omission control and inspect detection and blocking steps before assigning it to TOE1 (standard IHC practice; UniProt Q96GM8: nuclear localization). |
| No nuclear signal in a reported High cell population. | First check that the expected cells are present and that the detection run worked (standard IHC practice). An absent result disagrees with HPA's High observation for that population, but one failed section does not establish loss of TOE1 expression (HPA tissue IHC: reported High cells; Supported, pending external verification). |
💡Expected TOE1 appearanceCall a section positive when identifiable cells show predominantly nuclear chromogen, preferably in a reported High population such as adipocytes or bone marrow hematopoietic cells (HPA tissue IHC); broad cytoplasmic or extracellular color without matching nuclear signal is suspect (UniProt Q96GM8: nuclear localization; standard IHC practice).
| Compartment and tissue distribution | TOE1 is nuclear, including nucleolar and nuclear-speckle annotations, and is widely expressed (UniProt Q96GM8). HPA reports general nuclear tissue staining with different cell-level intensities (HPA tissue IHC). Use the named cells when judging a section; wide expression alone does not predict uniform intensity in every cell. |
| Strength of the IHC evidence | The tissue profile is Supported because staining and RNA expression are highly consistent, with external verification pending (HPA tissue IHC: reliability description). Of the listed antibodies, HPA069119 has Supported IHC validation; HPA053775 has no IHC status in this payload (HPA antibodies). Do not transfer the former's IHC status to the latter. |
| Isoforms and processing | UniProt lists 2 isoforms and a chain spanning residues 2–510, with no signal peptide or propeptide reported (UniProt Q96GM8). These annotations do not identify the antibody epitope or establish isoform-specific staining. Interpret compartment and cell distribution without claiming that a particular isoform explains a staining difference. |
| Retrieval and detection controls | For paraffin IHC, optimize retrieval and check detection with a reported positive population and suitable controls (standard IHC practice). No TOE1-specific retrieval condition, fixation effect or dilution is supplied. Endogenous detection activity can produce misleading color, so a control lacking primary antibody helps investigate diffuse or unexpected signal (standard IHC practice). |
| IF/ICC Q&A: where should fluorescence appear? | Mainly in nucleoplasm, with possible nuclear-body signal; both HPA subcellular assignments are marked uncertain (HPA ICC-IF). UniProt also annotates nucleolus and nuclear speckles (UniProt Q96GM8). HPA lists Enhanced ICC validation for both antibodies (HPA antibodies). These IF/ICC observations do not supply an IHC protocol or override the tissue IHC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Reported High cells are present, but nuclei remain blank. | The section or detection run may have failed; absence in one section alone is inconclusive (standard IHC practice; HPA tissue IHC: High cell observations). | Confirm cell identity on the counterstain, inspect a processed positive section and detection controls, then reassess the nuclear signal (standard IHC practice). |
| Color concentrates in cytoplasm or at cell borders. | The distribution conflicts with nuclear TOE1 and its lack of a transmembrane segment (UniProt Q96GM8); nonspecific staining is possible (standard IHC practice). | Compare with a reported High cell population and a primary-omission control; score TOE1 only where the compartment assignment is credible (HPA tissue IHC; standard IHC practice). |
| Strong staining appears in parathyroid glandular cells or liver cholangiocytes. | HPA calls these populations Low, so a strong result needs scrutiny; the payload does not classify them as negative (HPA tissue IHC). | Check the named cells, nuclear localization and control staining before attributing the signal to TOE1 (HPA tissue IHC: general nuclear expression; standard IHC practice). |
| Diffuse color obscures nuclear boundaries. | Background from blocking or detection steps can hinder localization calls, including through endogenous detection activity (standard IHC practice). | Inspect a primary-omission control and review blocking, detection and counterstain conditions; repeat scoring only when nuclei can be distinguished (standard IHC practice). |
| Two antibodies give different IHC staining patterns. | The listed antibodies have different recorded IHC statuses: HPA069119 is Supported, while HPA053775 has no IHC status here (HPA antibodies). | Record which antibody produced each pattern and compare nuclear staining in the same reported High cells; do not cite ICC validation as IHC validation (HPA antibodies; HPA tissue IHC). |
| An IF image shows nuclear bodies, while IHC looks broadly nuclear. | HPA's ICC-IF nuclear-body and nucleoplasm assignments are uncertain, whereas its tissue IHC profile is general nuclear expression (HPA subcellular; HPA tissue IHC). | Interpret the paraffin section against its cell-level IHC profile and counterstain; reserve detailed subnuclear comparisons for the separate IF/ICC assessment (HPA tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing TOE1 staining
A09852-2 · IHCIHC analysis of TOE1 using anti-TOE1 antibody (A09852-2). TOE1 was detected in a paraffin-embedded section of human cerebellum tissue. Heat
A09852-2 · IFIF analysis of TOE1 using anti-TOE1 antibody (A09852-2) and anti-Tubulin Alpha antibody (M03989-3). TOE1 was detected in immunocytochemical
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000132773-TOE1/tissue/cerebral+cortex#imid_19013001
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000132773-TOE1/tissue/colon#imid_19012919
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000132773-TOE1/tissue/liver#imid_19012920
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000132773-TOE1/tissue/kidney#imid_19012929
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA053775; green = TOE1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA053775; green = TOE1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TOE1 IHC & IF
🧪Run adipose tissue first and score TOE1 staining in adipocyte nuclei (HPA: High in adipocytes; UniProt Q96GM8: nuclear localization). HPA detects TOE1 in all 45 scored tissues, so there is no supported negative tissue or validated internal negative cell population; cells that appear negative should show counterstain without specific nuclear signal and require independent validation, while the no-primary and isotype slides carry the negative controls (HPA: no negative tissues).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: TOE1 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 TOE1 in A-549, HeLa, U2OS, CACO-2, RT-4, KOLF2.1J, NIH 3T3, with annotated localisation: Nucleoplasm (uncertain) (HPA subcellular).
Technical controls: Include a no-primary slide with the secondary and detection reagents, an isotype-matched rabbit immunoglobulin control, and TOE1-knockout tissue or cells if available as a biological negative (caption: rabbit anti-TOE1 antibody; standard IHC control practice). Quench endogenous peroxidase for HRP/DAB detection, and score the peripheral adipocyte nuclei rather than empty lipid spaces (caption: HRP/DAB detection; HPA: High in adipocytes).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A09852-2 tissue-IHC caption does not state the fixative (caption: paraffin-embedded human cerebellum; fixative unreported). That caption demonstrates heat-mediated retrieval in EDTA at pH 8.0 before chromogenic staining, but does not establish whether retrieval is required; frozen-section and IF suitability are not established by this IHC caption (caption: EDTA retrieval and HRP/DAB detection). In adipose tissue, identify the thin peripheral nuclei carefully so empty lipid spaces are not scored as negative cells (HPA: High in adipocytes; standard adipose histology).
HPA tissue IHC evidence for TOE1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — High consistency between antibody staining and RNA expression data. Pending external verification.). 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: TOE1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced TOE1 IHC Tips
Troubleshoot TOE1 staining in paraffin sections by checking retrieval, nuclear localisation, background and scoring against the catalog antibody’s tissue result and reference profiles.
Which retrieval conditions should I start with for TOE1 in paraffin sections?
Start with heat mediated retrieval in EDTA at pH 8.0 (datasheet A09852-2). The selected paraffin section result used this condition before overnight incubation with 2 μg/ml antibody at 4°C (datasheet A09852-2). If nuclear staining is weak, compare a longer heat exposure on adjacent sections while keeping cooling, antibody incubation and detection constant (general IHC practice). Examine tissue integrity alongside signal, because excessive retrieval can damage morphology and complicate nuclear scoring (general IHC practice). Record retrieval duration and heating method for each run; the caption identifies the buffer and pH but gives no heating time (datasheet A09852-2).
How should I troubleshoot fixation when TOE1 nuclear staining is inconsistent?
The selected tissue caption identifies a paraffin section but does not state its fixative, so target specific fixation sensitivity is unknown (datasheet A09852-2). Record the fixative, time to fixation and fixation duration for each specimen before comparing TOE1 intensity (general IHC practice). Where processing records differ, stain adjacent sections in one run with the same pH 8.0 EDTA retrieval and detection conditions (datasheet A09852-2; general IHC practice). Compare nuclear signal with morphology and a processing control; weak staining in poorly preserved regions alone does not establish TOE1 loss (general IHC practice). Do not infer a fixation effect from TOE1 topology, modifications or tissue staining patterns (UniProt Q96GM8; HPA tissue IHC).
What staining pattern should count as plausible TOE1 localisation?
Prioritise nuclear staining: UniProt places TOE1 in the nucleolus and nuclear speckles, while HPA reports general nuclear tissue expression (UniProt Q96GM8; HPA tissue IHC). HPA’s cell imaging describes mainly nucleoplasmic staining with additional nuclear bodies, but labels those subcellular assignments uncertain (HPA subcellular). Inspect counterstained nuclei at high magnification before scoring puncta, since a chromogenic section cannot by itself resolve every nuclear structure (general IHC practice). Diffuse staining outside nuclei warrants checks of background and detection controls before assignment to TOE1 (UniProt Q96GM8; general IHC practice). Compare staining within the same cell class across sections, because HPA reports different levels among sampled tissue and cell types (HPA tissue IHC).
Could isoforms or epitope accessibility explain discordant TOE1 staining?
TOE1 has 2 annotated isoforms, but the supplied antibody caption does not map its epitope or establish isoform recognition (UniProt Q96GM8; datasheet A09852-2). Treat differences between specimens as unresolved until the antibody’s immunogen or epitope information can be checked against both isoforms (UniProt Q96GM8; general IHC practice). Nuclear accessibility may change with section processing, so compare adjacent sections using the documented pH 8.0 EDTA retrieval first (datasheet A09852-2; general IHC practice). TOE1 has annotated phosphorylation sites at positions 5, 358 and 428, but their effect on this antibody is unknown (UniProt Q96GM8; datasheet A09852-2). Avoid assigning isoform or modification status from chromogenic intensity alone (general IHC practice).
How can I compare TOE1 immunofluorescence with this chromogenic IHC result?
Plan IF as a separate assay: the selected antibody evidence describes paraffin section chromogenic IHC, with no IF fixation or permeabilisation conditions supplied (datasheet A09852-2). Multiplex TOE1 with an independently validated marker for the expected cell class; Purkinje cells in cerebellum are one documented high staining reference (HPA tissue IHC; general IF practice). Choose a fluorophore channel after checking tissue autofluorescence and single stain controls, then compare nuclear signal with a DNA counterstain (general IF practice). TOE1 has no annotated transmembrane segment and is reported in nuclear compartments, so use nuclear access as the permeabilisation goal while confirming the antibody epitope’s accessibility experimentally (UniProt Q96GM8; general IF practice).
What should I check when TOE1 DAB staining appears widespread?
First compare staining with the expected nuclear distribution and with a no primary antibody control (UniProt Q96GM8; general IHC practice). The selected protocol used 10% goat serum, a peroxidase conjugated secondary antibody and DAB; these document its blocking and detection conditions, not a background rate (datasheet A09852-2). Include a peroxidase block and assess a detection only control when diffuse brown signal persists, because endogenous enzyme activity can contribute to chromogenic background (general IHC practice). Check whether background follows damaged edges, folds or poorly preserved regions rather than intact nuclei (general IHC practice). Adjust antibody concentration or detection exposure one variable at a time against the documented 2 μg/ml incubation (datasheet A09852-2; general IHC practice).
How should I quantify TOE1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell class and nuclear scoring threshold before comparing slides; HPA reports general nuclear expression and high staining in several sampled cell classes (HPA tissue IHC). Score the percentage of positive nuclei and intensity, or calculate an H-score from 0–300 when intensity grades are reproducible (general IHC practice). Normalise counts to evaluable nuclei within the same annotated cell class, or report positive nuclei per mm² of viable tissue with area recorded (general IHC practice). Exclude folds, necrosis and section edges using the same rules for every specimen (general IHC practice). Keep retrieval, detection and image settings matched, and report biological replicates separately from technical sections (general IHC practice).
How do I distinguish genuine TOE1 staining from a tissue artefact?
A plausible positive should occupy nuclei in intact cells, consistent with UniProt nuclear localisation and HPA’s general nuclear tissue pattern (UniProt Q96GM8; HPA tissue IHC). In cerebellum, high staining in Purkinje cells provides a documented cell class to inspect, while HPA also reports low staining in liver cholangiocytes (HPA tissue IHC). Brown signal confined to cytoplasm, section edges or necrotic regions needs background and morphology review before interpretation (UniProt Q96GM8; general IHC practice). Compare a no primary control and the peroxidase block when staining appears independent of nuclear boundaries (general IHC practice). HPA rates tissue staining as Supported, with external verification pending, so use independent controls before claiming biological differences (HPA tissue IHC).
Boster reagents
Best TOE1 / Target of EGR1 protein 1 IHC Antibodies
A09852-2 has IHC data from a paraffin-embedded human cerebellum section and IF/ICC data from U2OS cells (A09852-2 IHC/IF image captions); listed reactivity is human (catalog: Human).
Real IHC data
Anti-TOE1 Antibody ®
Cat # A09852-2
A09852-2 will render with IHC data from a paraffin-embedded human cerebellum section (A09852-2 IHC image caption). The same SKU is listed for IF and ICC and has IF data from U2OS cells (catalog: applications; A09852-2 IF image caption).
Which to pick: For tissue IHC, choose A09852-2: its own caption documents heat retrieval in EDTA at pH 8.0 and 2 μg/ml primary antibody on a paraffin-embedded human cerebellum section; the fixative is unreported (A09852-2 IHC image caption). For IF/ICC, A09852-2 is also listed for both applications and has a U2OS-cell IF image (catalog: applications; A09852-2 IF image caption). No cross-species or clone-specific choice is supported by this catalog entry (catalog: Human reactivity; clone unreported).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96GM8 (TOE1_HUMAN, Target of EGR1 protein 1).
- Human Protein Atlas. TOE1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. TOE1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nuclear bodies..
- Human Protein Atlas. TOE1 antibody validation summary (2 antibodies).
- Distinct Poly(A) nucleases have differential impact on sut-2 dependent tauopathy phenotypes. Neurobiology of disease 2021 — PMC8092974.
- Transcriptomic analysis reveals the potential role of TOE1 in hepatocellular carcinoma. Scientific reports 2025 — PMC12764802.
- PubMed PMID:12562764 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TOE1 (Target of EGR1 protein 1). Expected localisation General nuclear staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A09852-2). Validated IHC antibody: A09852-2 at 2-5μg/ml (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 cerebellum tissue; fixative not specified (datasheet A09852-2); 2. Fixation: Image fixative and duration unreported (datasheet A09852-2); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A09852-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A09852-2); 8. Primary antibody: Rabbit anti-TOE1, 2-5μg/ml (datasheet A09852-2); 9. Primary incubation: Overnight at 4 °C (datasheet A09852-2). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A09852-2); Caveat: Cholangiocytes may stain weakly (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt); Regulation: Widely expressed; no regulator annotated (UniProt). Evidence: UniProt Q96GM8, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A09852-2, PMC8092974, PMC12764802. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TOE1/immunohistochemistry-ihc-guide.