This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
TXNDC5 / Thioredoxin domain-containing protein 5 · IHC design guide
Design Immunohistochemistry for TXNDC5
Plan paraffin IHC for TXNDC5 using the cytoplasmic staining reported in exocrine pancreatic cells and leukocyte subsets (HPA tissue IHC). Start with the catalog antibody at 2–5 μg/ml (datasheet A05772-1) and include cardiomyocytes as a comparison where staining was not detected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TXNDC5 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 staining in tissue (HPA tissue IHC) | |
| Staining pattern | Cytoplasm of leukocyte subsets and exocrine pancreatic cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05772-1) | |
| Positive control | Duodenum+4 more · see all | |
| Negative control | Caudate+4 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) | |
| Caveat | Positive cell subsets may be unevenly distributed (HPA tissue IHC) | |
| Regulation | No expression regulation annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; verify epitope coverage for both (UniProt) |
Section 1
Recommended TXNDC5 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA pH 8.0 retrieval (datasheet: A05772-1). The published protocols below cover gastric mucosa, colon biopsies, and mouse ankle joints (PMC4315038; PMC11786489; PMC3541148).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A05772-1) |
| Fixation | Image fixative and duration unreported (datasheet A05772-1); 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 A05772-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05772-1) |
| Primary antibody | Rabbit anti-TXNDC5, 2-5 μg/ml (datasheet A05772-1) |
| Primary incubation | Overnight at 4 °C (datasheet A05772-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A05772-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TXNDC5-positive staining in paneth cells of duodenum (HPA tissue IHC: High). HPA tissue profile: cytoplasmic expression in several tissue types, most abundant in a subset of leukocytes, exocrine pancreas and a subset of decidual cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet: A05772-1). For mouse ankle joints, a published alternative is citrate retrieval at 95°C for 10 min (PMC3541148).
Section 2
What Is the Expected TXNDC5 Staining Pattern?
TXNDC5 is an endoplasmic reticulum (ER) lumen protein with no transmembrane segment (UniProt Q8NBS9 topology). In paraffin IHC, expect cytoplasmic staining, especially in pancreatic exocrine glandular cells, intestinal Paneth and goblet cells, and tonsillar non-germinal center cells (HPA: High in each). HPA rates tissue staining Enhanced, while reporting medium consistency with RNA expression; interpret the pattern alongside cell identity (HPA: tissue IHC reliability).
| Granular or reticular cytoplasmic staining in pancreatic exocrine cells or intestinal Paneth cells. | This fits an ER-associated pattern in cells reported High by tissue IHC (UniProt Q8NBS9 localization; HPA: High in both). Compare the signal with neighboring cell types on the same section before calling the tissue uniformly positive. |
| Predominantly nuclear, sharply membranous, or extracellular staining. | Treat this as an unexpected compartment pattern: TXNDC5 is ER-luminal and has no transmembrane segment (UniProt Q8NBS9 topology). Review morphology and detection controls before assigning it to TXNDC5; HPA reports cytoplasmic tissue expression (HPA: tissue IHC profile). |
| Strong signal in cells reported as not detected, such as cardiomyocytes. | This conflicts with the listed cell-level reference (HPA: Not detected in cardiomyocytes). Check for cross-reactivity or endogenous chromogenic activity using appropriate controls (general IHC practice). A tissue name alone does not identify which cells produced the signal. |
| Broad, uniform color over cells and surrounding tissue, obscuring cell boundaries. | This is difficult to score as cellular TXNDC5 staining. Assess background with a primary-omission control and inspect blocking, washing, and detection steps (general IHC practice). The reference pattern is cytoplasmic and cell selective (HPA: tissue IHC profile). |
| No visible staining in pancreatic exocrine cells on an otherwise interpretable section. | This fails a listed high-expression reference (HPA: High in pancreatic exocrine glandular cells). Confirm that the expected cells are present, then review the IHC detection workflow and antibody validation before concluding that TXNDC5 is absent (general IHC practice). |
💡Expected TXNDC5 appearanceCall a positive result when cytoplasmic, ER-like staining is clear in listed High cell populations, such as pancreatic exocrine or intestinal Paneth cells (UniProt Q8NBS9 localization; HPA: High in both); dominant nuclear or extracellular color is suspect (UniProt Q8NBS9 localization).
| Compartment and processing | TXNDC5 enters the secretory pathway through a signal peptide at residues 1–32; the annotated chain spans 33–432 (UniProt Q8NBS9 processing). Its ER-lumen location supports a cytoplasmic IHC pattern, but the payload gives no antibody epitope, so processing cannot predict antibody-specific staining. |
| Cell-level reference strength | Tissue IHC is rated Enhanced, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability). HPA lists High staining in selected cells, not every cell within those tissues (HPA: tissue IHC profile). Use cell identity when comparing sections. |
| Isoforms and target scope | Two isoforms are annotated (UniProt Q8NBS9 isoforms). Their epitopes and tissue-specific staining are not supplied, so the isoform count alone cannot explain a positive, weak, or absent IHC result. Keep an isoform-based explanation provisional until antibody-specific evidence is available. |
| IF/ICC Q&A: What pattern should I expect? | An ER-associated intracellular pattern is the supported expectation: HPA approves the ER as the main ICC-IF location, and UniProt places TXNDC5 in the ER lumen (HPA: subcellular; UniProt Q8NBS9 localization). This informs interpretation only; IF/ICC procedures belong in its separate guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| Pancreatic exocrine cells show no signal. | The expected High cell population may be absent from the field, or the IHC workflow may have failed (HPA: High in pancreatic exocrine glandular cells; general IHC practice). | Verify cell identity and section quality; review the antibody's IHC validation, detection reagents, and control results before interpreting the negative (general IHC practice). |
| Color is mainly nuclear or outlines cell membranes. | The dominant compartment conflicts with an ER-luminal protein lacking a transmembrane segment (UniProt Q8NBS9 topology). | Recheck counterstain and cell boundaries, then compare with a primary-omission control and a listed High cell population (general IHC practice; HPA: High in pancreatic exocrine cells). |
| Cardiomyocytes stain strongly. | That cell-level result conflicts with the HPA reference and may reflect cross-reactivity or endogenous detection activity (HPA: Not detected in cardiomyocytes; general IHC practice). | Inspect a primary-omission control and the detection chemistry; confirm that the colored cells are cardiomyocytes before scoring TXNDC5 (general IHC practice). |
| Diffuse chromogen masks cellular detail. | Background may arise from endogenous detection activity or incomplete blocking or washing (general IHC practice); the diffuse appearance does not establish TXNDC5 localization. | Compare a primary-omission control, review blocking and washes, and score only clearly cellular staining against the cytoplasmic reference (general IHC practice; HPA: tissue IHC profile). |
| Only some cells within a positive tissue stain. | HPA reports expression in specific populations, including intestinal Paneth and goblet cells, rather than a tissue-wide uniform pattern (HPA: High in Paneth and goblet cells). | Identify the stained cell types and compare them with the appropriate HPA cell-level entry before calling the section inconsistent (general IHC practice; HPA: tissue IHC profile). |
| A weak section raises concern about fixation or retrieval. | The supplied sources give no TXNDC5-specific fixation sensitivity or retrieval condition; HPA staining levels and UniProt topology do not establish one (HPA: tissue IHC; UniProt Q8NBS9 topology). | Review the catalog antibody's IHC-P instructions and run suitable positive and detection controls when optimizing the general paraffin IHC workflow (general IHC practice). |
Experiment evidence
IHC & IF images showing TXNDC5 staining
A05772-1 · IHCIHC analysis of TXNDC5 using anti-TXNDC5 antibody (A05772-1). TXNDC5 was detected in a paraffin-embedded section of human breast cancer tiss
A05772-1 · IHCIHC analysis of TXNDC5 using anti-TXNDC5 antibody (A05772-1). TXNDC5 was detected in a paraffin-embedded section of human larynx squamous ce
A05772-1 · IFIF analysis of TXNDC5 using anti-TXNDC5 antibody (A05772-1). TXNDC5 was detected in a paraffin-embedded section of human colon cancer tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000239264-TXNDC5/tissue/cerebral+cortex#imid_12118795
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000239264-TXNDC5/tissue/colon#imid_12118729
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000239264-TXNDC5/tissue/liver#imid_12118730
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000239264-TXNDC5/tissue/kidney#imid_12118739
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA034678; green = TXNDC5, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA034678; green = TXNDC5, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TXNDC5 IHC & IF
🧪Run pancreas first: exocrine glandular cells should stain (HPA: High in pancreatic exocrine glandular cells). Use heart muscle cardiomyocytes as the negative tissue (HPA: Not detected in cardiomyocytes); on the pancreas slide, background regions should lack specific chromogen deposits, but other cell types should not be assumed TXNDC5-negative (standard IHC practice).
Positive control tissue: Duodenum (Paneth cells, HPA High)
Negative control tissue: Caudate (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TXNDC5 in A-431, U-251MG, U2OS, with annotated localisation: Endoplasmic reticulum (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit immunoglobulin isotype control appropriate to the primary antibody’s clonality (selected-SKU caption: rabbit primary; standard IHC practice). Confirm specificity with TXNDC5 knockout material or a validated immunizing-peptide block, and quench endogenous peroxidase when using HRP/DAB detection (selected-SKU caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption leaves the fixative unreported (selected-SKU caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required or that this condition is optimal (selected-SKU caption: EDTA heat retrieval). Paraffin IHC has a documented starting condition; whether frozen sections or IF/ICC are easier is unreported, and prompt handling matters for autolysis-prone pancreas tissue (selected-SKU caption: paraffin IHC; standard histology practice).
HPA tissue IHC evidence for TXNDC5
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 | Paneth cells | High | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Medium | Protein (IHC) | HPA → |
| Pancreas | Exocrine glandular cells | High | Protein (IHC) | HPA → |
| Small intestine | Goblet cells | High | Protein (IHC) | HPA → |
| Stomach | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TXNDC5 IHC Tips
Troubleshoot TXNDC5 staining in paraffin sections by checking retrieval, cellular distribution and controls before interpreting chromogenic signal.
What retrieval conditions should I try first for weak TXNDC5 staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A05772-1). The catalog antibody stained a human breast cancer section after this retrieval, followed by 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A05772-1). If staining remains weak, vary heating time on matched sections while keeping antibody concentration and detection constant (standard IHC practice). Compare signal in exocrine pancreatic cells or Paneth cells with a no-primary control, and reject conditions that increase diffuse background along with signal (HPA: High in both cell types; standard IHC practice).
Could fixation explain weak or uneven TXNDC5 staining?
Target-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not state a fixative, and no TXNDC5 fixation comparison is supplied (datasheet A05772-1). Record the fixative, time to fixation and fixation duration for each specimen, then compare sections processed together before changing retrieval or antibody concentration (standard IHC practice). For uneven staining, inspect section thickness, folds, detached tissue and drying marks alongside fixation history (standard IHC practice). Use the documented EDTA pH 8.0 retrieval as the starting condition, and treat any fixation adjustment as an experiment requiring matched controls (datasheet A05772-1; standard IHC practice).
Where should convincing chromogenic TXNDC5 signal appear within cells?
Expect an intracellular, predominantly cytoplasmic pattern consistent with an endoplasmic reticulum lumen protein that has no transmembrane segment (UniProt Q8NBS9 topology; HPA: cytoplasmic tissue expression). The HPA subcellular assessment places TXNDC5 in the endoplasmic reticulum, so sharply isolated nuclear or cell-surface staining needs separate validation (HPA: approved endoplasmic reticulum location; UniProt Q8NBS9 topology). Assess localisation within morphologically intact cells before assigning a positive score (standard IHC practice). Compare high-expression reference cells, such as exocrine pancreatic cells, with adjacent low-staining compartments and a no-primary section under the same DAB exposure (HPA: High in exocrine glandular cells; standard IHC practice).
How can epitope uncertainty affect interpretation of TXNDC5 IHC?
TXNDC5 has 2 annotated isoforms, but the supplied antibody caption does not map the recognized epitope or establish isoform coverage (UniProt Q8NBS9 isoforms; datasheet A05772-1). The precursor has a signal peptide at residues 1–32 and a mature chain at 33–432, making epitope position relevant when evaluating antibody specificity (UniProt Q8NBS9 processing). Its 3 thioredoxin domains span residues 36–169, 170–295 and 304–429 (UniProt Q8NBS9 domains). Obtain the immunogen or epitope information before claiming isoform-specific staining, and corroborate disputed patterns with an independently validated antibody if available (standard IHC validation practice).
How should I investigate TXNDC5 localisation by multiplex IF?
For a separate IF experiment, pair TXNDC5 with a marker for the cell population being assessed, such as a validated exocrine pancreatic cell marker when examining pancreas (HPA: High in exocrine glandular cells; standard IF practice). Choose spectrally separated fluorophores and inspect an unstained section to select channels with less tissue autofluorescence (standard IF practice). Because TXNDC5 is luminal within the endoplasmic reticulum, permeabilise cells sufficiently for antibody access and compare signal with a no-primary control (UniProt Q8NBS9 topology; standard IF practice). Assess intracellular overlap and cell identity independently; the HPA reports an approved endoplasmic reticulum location from ICC/IF images (HPA: subcellular location).
What causes widespread brown signal in a TXNDC5 IHC section?
First compare the stained section with a no-primary control and inspect pigment, folds and section edges before attributing widespread DAB to TXNDC5 (standard IHC practice). Apply an endogenous peroxidase block before HRP detection and keep DAB development time consistent across sections (standard chromogenic IHC practice). The catalog image used 10% goat serum blocking and an HRP/DAB detection workflow, which provides starting conditions for investigating nonspecific staining (datasheet A05772-1). If background persists, titrate the 2 μg/ml primary concentration downward on matched sections and verify that expected intracellular staining remains in a positive reference tissue (datasheet A05772-1; standard IHC practice).
How should I score TXNDC5 across heterogeneous tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region of interest before scoring, because reported TXNDC5 staining varies among cell types within tissues (HPA: tissue IHC profile; standard IHC practice). For identifiable cells, report the percentage positive and an H-score using the same intensity thresholds across cases; for dispersed positive cells, report density per mm² (standard IHC scoring practice). Normalise cell counts to viable cells of the prespecified population or density to viable tissue area, excluding necrosis and folds (standard IHC practice). Keep retrieval at EDTA pH 8.0 and DAB development consistent across the comparison set, and document interobserver agreement where scoring is subjective (datasheet A05772-1; standard IHC practice).
Which findings distinguish genuine TXNDC5 staining from artefact?
Give greatest weight to intracellular staining in intact, expected cells: TXNDC5 resides in the endoplasmic reticulum lumen, and HPA reports high staining in exocrine pancreatic and Paneth cells (UniProt Q8NBS9 topology; HPA: tissue IHC profile). Isolated nuclear rims, extracellular deposits or uniform cell-surface outlines conflict with the annotated compartment and warrant validation (HPA: approved endoplasmic reticulum location; standard IHC interpretation). Check whether apparent positivity is confined to tissue edges or necrotic areas, and whether it persists in a no-primary control after peroxidase blocking (standard IHC practice). Interpret a clean positive reference alongside a reported negative cell population, such as cardiomyocytes, while allowing for specimen-specific variation (HPA: cardiomyocytes Not detected; standard IHC practice).
Boster reagents
Best TXNDC5 / Thioredoxin domain-containing protein 5 IHC Antibodies
The IHC-validated A05772-1 has paraffin-section IHC images from human breast, larynx, prostate, and rectum cancers, plus a paraffin-section IF image from human colon cancer (catalog image captions).
Real IHC data
Anti-TXNDC5 Antibody ®
Cat # A05772-1
A05772-1 is listed for IHC in human, mouse, and rat; its IHC images show paraffin-embedded human breast, larynx, prostate, and rectum cancer sections (catalog: applications, reactivity, IHC image captions). Its IF image shows a paraffin-embedded human colon cancer section (catalog: IF image caption).
Which to pick: For tissue IHC, choose A05772-1: its images document paraffin-section staining at 2 μg/ml, while the catalog recommends 2–5 μg/ml (IHC image captions; catalog: IHC dilution). For fluorescence work, the same SKU has paraffin-section IF data at 5 μg/ml; ICC is not listed, and clonality is unreported (IF image caption; catalog: applications, clone). For cross-species IHC, the catalog lists human, mouse, and rat reactivity, but the supplied IHC images show human tissue only; the fixative is unreported (catalog: reactivity; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8NBS9 (TXND5_HUMAN, Thioredoxin domain-containing protein 5).
- Human Protein Atlas. TXNDC5 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. TXNDC5 subcellular location (ICC-IF): Localized to the endoplasmic reticulum..
- Human Protein Atlas. TXNDC5 antibody validation summary (2 antibodies).
- An immunohistochemical study of thioredoxin domain-containing 5 expression in gastric adenocarcinoma. Oncology letters 2015 — PMC4315038.
- Quantitative proteomic studies of the intestinal mucosa provide new insights into the molecular mechanism of ulcerative colitis. BMC gastroenterology 2025 — PMC11786489.
- Investigate pathogenic mechanism of TXNDC5 in rheumatoid arthritis. PloS one 2013 — PMC3541148.
- ALKBH5-mediated m6A demethylation of TXNDC5 drives malignant progression in gastric cancer. Epigenomics 2026 — PMC12826721.
- PubMed PMID:12975309 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TXNDC5 (Thioredoxin domain-containing protein 5). Expected localisation Cytoplasmic staining in tissue (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05772-1). Validated IHC antibody: A05772-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Duodenum (Paneth cells); negative Caudate. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A05772-1); 2. Fixation: Image fixative and duration unreported (datasheet A05772-1); 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 A05772-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05772-1); 8. Primary antibody: Rabbit anti-TXNDC5, 2-5 μg/ml (datasheet A05772-1); 9. Primary incubation: Overnight at 4 °C (datasheet A05772-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Positive cell subsets may be unevenly distributed (HPA tissue IHC); Isoform / epitope: 2 isoforms; verify epitope coverage for both (UniProt); Regulation: No expression regulation annotated (UniProt). Evidence: UniProt Q8NBS9, HPA tissue IHC · Enhanced, HPA ICC-IF · endoplasmic reticulum., Datasheet A05772-1, PMC4315038, PMC11786489. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TXNDC5/immunohistochemistry-ihc-guide.