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TXN / Thioredoxin · IHC design guide
Design Immunohistochemistry for TXN
Plan TXN staining in paraffin sections using kidney tubules as a high-staining reference (HPA tissue IHC). This guide covers fixation, cytoplasmic and nuclear staining, and radiation-dependent nuclear redistribution (standard IHC practice; HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TXN 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 | High in kidney tubules; cytoplasmic and nuclear (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Duodenum+4 more · see all | |
| Negative control | Adipose tissue+2 more · see all |
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 A01219) | |
| Caveat | Radiation can shift TXN into nuclei (UniProt) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; no transmembrane segment; epitope coverage unknown (UniProt) |
Section 1
Recommended TXN IHC & IF Protocols
Compare the catalog antibody protocol with three published TXN IHC protocols using paraffin sections (PMC11256699; PMC5438738; PMC4694848).
| Sample | Paraffin-embedded H. esophagus tissue; fixative not specified (datasheet A01219) |
| Fixation | Image fixative and duration unreported (datasheet A01219); 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-TXN, 1:25 (datasheet A01219) |
| 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 | TXN-positive staining in glandular cells of duodenum (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic and nuclear expression. 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 setting); compare citrate pH 6.0 where needed (PMC11256699; PMC4694848).
Section 2
What Is the Expected TXN Staining Pattern?
TXN should show cytoplasmic and nuclear staining in paraffin sections, with strong signal in several specified cell types, including kidney tubular cells and intestinal glandular cells (HPA: general cytoplasmic and nuclear expression; High in these cells). TXN has no transmembrane segment, although it can be secreted (UniProt P10599 topology and subcellular location). HPA rates the tissue pattern Enhanced, with medium consistency between staining and RNA data (HPA: tissue IHC reliability).
| Cytoplasmic staining with variable nuclear staining in kidney tubular cells or small-intestinal glandular cells (HPA: High in both). | This fits the expected compartments and provides a strong positive comparison (HPA: general cytoplasmic and nuclear expression; High in both cell types). Compare cells within the same section before judging intensity (general IHC practice). |
| Signal outlines cell membranes while cytoplasm and nuclei remain unstained. | A membrane-only pattern conflicts with the reported cytoplasmic and nuclear tissue pattern and the absence of a transmembrane segment (HPA: tissue IHC profile; UniProt P10599 topology). Check morphology and detection controls for artefact (general IHC practice). |
| Strong staining appears in adipocytes or bone-marrow hematopoietic cells. | These cell types are reported as Not detected, so prominent signal warrants scrutiny for cross-reactivity or endogenous detection activity (HPA: adipocytes and hematopoietic cells Not detected; general IHC practice). The HPA categories are reference observations, not universal negatives. |
| Diffuse color covers tissue and empty spaces, obscuring cell boundaries. | A slide-wide haze prevents reliable compartment or cell-type scoring (general IHC practice). It cannot establish TXN expression from the HPA pattern; assess background with the same detection system and an omitted-primary control (general IHC practice). |
| No cellular signal appears in kidney tubules or duodenal glandular cells. | Both are reported High and can flag an unsuccessful run, although any individual section may vary (HPA: High in kidney tubular and duodenal glandular cells). Check tissue preservation and assay controls before interpreting the specimen as TXN negative (general IHC practice). |
💡Expected TXN appearanceCall a section positive when recognizable cells show cytoplasmic staining, with possible nuclear staining, and strong signal in an HPA High cell type; isolated membrane outlines or uniform haze should prompt artefact checks (HPA: tissue IHC profile and High cell types; UniProt P10599 topology; general IHC practice).
| Cell choice for interpretation | Kidney tubular and duodenal glandular cells are High; adipocytes are Not detected (HPA: tissue IHC). Use these cell identities when comparing staining, while allowing for section-to-section variation (general IHC practice). |
| Nuclear distribution | TXN can move from cytoplasm to nucleus after PMA induction or radiation; nonirradiated cells are predominantly cytoplasmic (UniProt P10599 subcellular location). Record treatment context when nuclear intensity differs. |
| Topology and secretion | TXN lacks a transmembrane segment and is reported to use a leaderless secretory pathway (UniProt P10599 topology and subcellular location). Neither fact alone identifies an extracellular chromogenic deposit as specific staining. |
| IHC antibody evidence | HPA047478, HPA055752 and CAB008678 each have Enhanced IHC validation (HPA: antibody validation). That status supports the reported pattern but does not validate an unrelated antibody or every staining condition. |
| Isoforms and antigen region | Two isoforms are listed, and the annotated thioredoxin domain spans residues 2–105 (UniProt P10599 isoforms and domain). The payload gives no antibody epitope, so isoform recognition cannot be assigned. |
| IF/ICC Q&A: Where should signal appear? | Mainly in nucleoplasm and cytosol (HPA: ICC-IF subcellular summary). This supports a localization comparison; the IHC tissue intensity categories do not establish IF/ICC intensity or a protocol (HPA: tissue IHC; ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| A positive-control section has no visible cellular stain. | The run may have failed; kidney tubular cells are reported High (HPA: kidney tissue IHC; general IHC practice). | Verify tissue identity, primary application, detection reagents and the run control before scoring samples (general IHC practice). |
| Most tissue and empty spaces carry diffuse chromogen. | Nonspecific binding, overdevelopment or detection background can obscure localization (general IHC practice). | Compare an omitted-primary control, review blocking and detection steps, and shorten development if appropriate (general IHC practice). |
| Unexpected cells stain strongly in a reference-low or Not detected area. | Cell misidentification, cross-reactivity or endogenous detection activity may explain the signal (HPA: tissue IHC categories; general IHC practice). | Recheck morphology and controls; if using peroxidase detection, assess endogenous peroxidase background (general IHC practice). |
| Only sharp membrane outlines are visible. | This conflicts with the reported cellular distribution and lack of a transmembrane segment (HPA: tissue IHC profile; UniProt P10599 topology). | Inspect a known-positive cell type and detection controls before assigning membrane staining to TXN (HPA: High cell types; general IHC practice). |
| Nuclear staining differs between sample groups. | Stimulus-dependent nuclear translocation is documented after PMA induction and radiation (UniProt P10599 subcellular location). | Record treatment and compare matched samples; score cytoplasmic and nuclear compartments separately (UniProt P10599 subcellular location; general IHC practice). |
| An alternative primary gives a different pattern. | The listed antibodies have Enhanced IHC validation, but the supplied record gives no epitope or validation for the alternative (HPA: antibody validation; UniProt P10599 record). | Compare documented application validation and matched controls before attributing the discrepancy to TXN biology (general IHC practice). |
Experiment evidence
IHC & IF images showing TXN staining
A01219 · IHCImmunohistochemical analysis of paraffin-embedded H. esophagus section using TXN Antibody (C-term) (Cat#A01219). A01219 was diluted at 1:25
A01219 · IHCImmunohistochemical analysis of paraffin-embedded H. stomach section using TXN Antibody (C-term) (Cat#A01219). A01219 was diluted at 1:25 di
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000136810-TXN/tissue/cerebral+cortex#imid_17224575
HPA · EsophagusHuman Protein Atlas IHC, Esophagus. Source: https://www.proteinatlas.org/ENSG00000136810-TXN/tissue/esophagus#imid_17224547
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000136810-TXN/tissue/colon#imid_17224517
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000136810-TXN/tissue/liver#imid_17224502
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody HPA047478; green = TXN, blue = nucleus, red = microtubules.
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody HPA047478; green = TXN, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TXN IHC & IF
🧪Run duodenum first; its glandular cells should stain for TXN (HPA: High in duodenal glandular cells). Use adipose tissue as the negative comparator (HPA: Not detected in adipocytes); on the duodenal slide, assess nearby nonglandular cells for background staining rather than assuming they are TXN negative (HPA: duodenal glandular cells are High).
Positive control tissue: Duodenum (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TXN in U-251MG, A-431, U2OS, NIH 3T3, with annotated localisation: Nucleoplasm (enhanced), Cytosol (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a host-matched, clonality-matched isotype control; use TXN knockout material or a validated immunizing-peptide block as a biological specificity control (standard IHC practice). Check endogenous peroxidase and, if using the caption’s biotinylated secondary with DAB, endogenous biotin in the duodenal section (A01219 tissue-IHC caption: biotinylated secondary and DAB; standard IHC practice).
⚠️Feasibility: A TXN-specific fixation window and retrieval dependency are unreported in the supplied evidence; the fixative is unreported in the A01219 paraffin-section caption (A01219 tissue-IHC caption: fixative not stated). The evidence does not establish whether frozen sections or IF are easier than paraffin IHC; IF interpretation can use nucleoplasmic and cytosolic localization as a reference (HPA subcellular: nucleoplasm and cytosol). For duodenal DAB staining, endogenous biotin may contribute background when a biotin-based detection system is used (A01219 tissue-IHC caption: biotinylated secondary; standard IHC practice).
HPA tissue IHC evidence for TXN
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (cell body) | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Placenta | Decidual cells | High | Protein (IHC) | HPA → |
Section 3
Advanced TXN IHC Tips
Troubleshoot TXN staining in paraffin sections by checking retrieval, compartment pattern, controls, and scoring before interpreting chromogenic signal.
What retrieval conditions should I start with for TXN in paraffin sections?
Start with Tris-EDTA at pH 9.0, heated to 95–98 °C for 20 min (page retrieval setting: nuclear antigen). Let sections cool in buffer, then apply the catalog antibody at 1:25 as a starting dilution because that dilution appears in its paraffin-section caption (caption: A01219). If staining is weak, compare a shorter or longer heating period on adjacent sections while holding antibody dilution and detection constant (standard IHC practice). Score both cytoplasmic and nuclear signal, since restricting assessment to nuclei could miss the documented pattern (HPA: general cytoplasmic and nuclear expression). Include a no-primary control to identify retrieval-related background (standard IHC practice).
How should I troubleshoot suspected fixation effects on TXN staining?
Target-specific fixation sensitivity is unknown because the selected paraffin-section caption does not report a fixative (caption: A01219). Record the fixative, fixation duration, section thickness, and storage conditions for each block before comparing TXN staining (standard IHC practice). On adjacent sections, keep the pH 9.0, 95–98 °C, 20 min retrieval setting constant while checking whether signal varies by block or section depth (page retrieval setting: nuclear antigen). Run the same antibody dilution and detection conditions across those sections; the caption documents 1:25 for A01219 in paraffin sections (caption: A01219). Treat any fixation effect as a finding from that controlled comparison, rather than inferring one from TXN localisation or modifications (UniProt P10599: localisation and modified residues).
Is nuclear staining plausible when TXN is expected in the cytoplasm?
Yes: general cytoplasmic and nuclear tissue staining is documented (HPA: tissue IHC profile), and nucleoplasm plus cytosol are the main reported subcellular locations (HPA: subcellular). Compare nuclear and cytoplasmic staining separately in the same cell population, using the counterstain to define nuclei (standard IHC practice). TXN can move into the nucleus after PMA induction or irradiation, so a nuclear shift can be biologically plausible (UniProt P10599: PubMed:9108029; PubMed:11118054). That mechanism alone does not establish why a particular specimen stains in its nuclei; examine matched controls and tissue condition first (standard IHC practice). A diffuse extracellular-only pattern merits scrutiny even though leaderless secretion is reported (UniProt P10599: PubMed:1332947).
Could the C-terminal antibody miss a TXN isoform or modified epitope?
The selected antibody is described as C-terminal, but the caption gives no residue-level epitope or isoform recognition data (caption: A01219). TXN has 2 annotated isoforms and a thioredoxin domain spanning residues 2–105 (UniProt P10599: isoforms and domain). Its listed modifications include acetyllysine at residue 39 and S-nitrosocysteine at residues 62 and 69 (UniProt P10599: modified residues). Those annotations cannot establish whether this antibody detects both isoforms or whether modification changes binding (caption: A01219; UniProt P10599: isoforms and modified residues). If staining conflicts with expected localisation, compare an independently mapped epitope or an orthogonal TXN assay on matched material (standard IHC validation practice).
How can IF help resolve an ambiguous TXN chromogenic pattern?
Use IF as a companion localisation check, since nucleoplasm and cytosol are the main reported TXN locations (HPA: subcellular). Multiplex TXN with a marker identifying the cell population under study, and include a nuclear counterstain to separate nuclear from cytosolic signal (standard IF practice). Choose fluorophores after inspecting unstained tissue for autofluorescence; shift the TXN channel away from the strongest background and include single-label controls (standard IF practice). TXN has no transmembrane segment, so use mild permeabilisation when assessing intracellular epitopes, then verify that compartment boundaries remain clear (UniProt P10599: topology and localisation; standard IF practice). Keep IF optimisation separate from the catalog antibody’s 1:25 paraffin-section IHC condition (caption: A01219).
What controls help distinguish TXN staining from chromogenic background?
Run no-primary and detection-only controls alongside TXN sections to expose reagent-derived deposits and endogenous enzyme signal (standard chromogenic IHC practice). For a peroxidase and DAB workflow, block endogenous peroxidase and compare its control before calling granular brown deposits positive (standard chromogenic IHC practice). The selected caption reports a biotinylated secondary and DAB, so evaluate whether its detection chain contributes background in the specimen being tested (caption: A01219). Check tissue edges, folds, and damaged areas against intact regions at the same exposure and counterstain (standard IHC practice). A convincing pattern should resolve to cells and compatible cytoplasmic or nuclear compartments (HPA: general cytoplasmic and nuclear expression).
How should I score mixed nuclear and cytoplasmic TXN staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then record nuclear and cytoplasmic staining separately (HPA: general cytoplasmic and nuclear expression; standard IHC scoring practice). For each compartment, report percentage of positive cells and intensity, or calculate an H-score from intensity categories 0–3 (standard IHC scoring practice). Normalise cell-based scores to the number of evaluable cells in that population, and area-based counts to sampled tissue area in mm² (standard IHC quantification practice). Apply identical thresholds, counterstain review, and exclusion rules across sections; omit folds, necrosis, and damaged edges (standard IHC scoring practice). Report the compartment-specific result because redistribution can change its meaning (UniProt P10599: PMA- and radiation-associated nuclear translocation).
When is a strong TXN signal likely to be artefactual?
Treat a well-defined cytoplasmic or nuclear cellular pattern as plausible, since both compartments are reported for TXN (HPA: tissue IHC profile; UniProt P10599: localisation). Check whether the stained cells fit the specimen: high signal is reported in kidney tubule cells, while adipocytes are listed as not detected (HPA: kidney and adipose tissue). Staining limited to section edges, folds, or necrotic areas needs confirmation in intact tissue (standard IHC interpretation practice). Brown deposits persisting in a no-primary control can indicate detection background, including endogenous enzyme activity in a peroxidase workflow (standard chromogenic IHC practice). Interpret nuclear enrichment with specimen context and controls, since induced nuclear translocation is documented but does not explain every specimen (UniProt P10599: PubMed:9108029; PubMed:11118054).
Boster reagents
Best TXN / Thioredoxin IHC Antibodies
A01219 has human paraffin-section IHC images (catalog IHC captions); M01219 lists human IHC and ICC/IF applications without image examples (catalog applications, reactivity, image alts).
Real IHC data
Anti-TXN Antibody (C-term)
Cat # A01219
A01219 shows IHC staining of paraffin-embedded human esophagus and stomach sections (catalog IHC image captions). M01219 is listed for human IHC and ICC/IF, with no IHC or IF image captions supplied (catalog applications, reactivity, image alts).
Which to pick: For tissue IHC, choose A01219, a rabbit polyclonal antibody with human paraffin-section images; the fixative is unreported (catalog host and IHC image captions). For IF/ICC, M01219 is the listed rabbit monoclonal, clone AOGC-20, with human ICC/IF reactivity but no IF image example (catalog host, clone, applications, reactivity, IF image alts). Neither SKU documents nonhuman reactivity, so neither is supported here for cross-species work (catalog reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P10599 (THIO_HUMAN, Thioredoxin).
- Human Protein Atlas. TXN tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. TXN subcellular location (ICC-IF): Mainly localized to the nucleoplasm and cytosol..
- Human Protein Atlas. TXN antibody validation summary (3 antibodies).
- Identification of thioredoxin-1 as a biomarker of lung cancer and evaluation of its prognostic value based on bioinformatics analysis. Frontiers in oncology 2023 — PMC9911911.
- Increased level of TXNIP and nuclear translocation of TXN is associated with end stage renal disease and development of multiplex renal tumours. BMC nephrology 2024 — PMC11256699.
- Correlation of five secretory proteins with the nasopharyngeal carcinoma metastasis and the clinical applications. Oncotarget 2017 — PMC5438738.
- Thioredoxin 1 mediates TGF-β-induced epithelial-mesenchymal transition in salivary adenoid cystic carcinoma. Oncotarget 2015 — PMC4694848.
- PubMed PMID:3170595 — UniProt-cited evidence.
- PubMed PMID:2785919 — UniProt-cited evidence.
- PubMed PMID:1874447 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TXN (Thioredoxin). 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: A01219 at 1:25 (the product's own IHC validation image is shown). Controls (HPA): positive Duodenum (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded H. esophagus tissue; fixative not specified (datasheet A01219); 2. Fixation: Image fixative and duration unreported (datasheet A01219); 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-TXN, 1:25 (datasheet A01219); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01219); Caveat: Radiation can shift TXN into nuclei (UniProt); Isoform / epitope: 2 isoforms; no transmembrane segment; epitope coverage unknown (UniProt); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt P10599, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm and cytosol., Datasheet A01219, PMC11256699, PMC5438738. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TXN/immunohistochemistry-ihc-guide.