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TSHZ3 / Teashirt homolog 3 · IHC design guide
Design Immunohistochemistry for TSHZ3
Plan chromogenic TSHZ3 IHC-P using the catalog antibody’s 2.5 μg/mL starting concentration (datasheet). Assess nuclear tissue staining while accounting for low concordance between antibody staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TSHZ3 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 | Ubiquitous nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | Nuclear staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| 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 A08453) | |
| Caveat | Staining has low concordance with RNA expression (HPA tissue IHC) | |
| Regulation | Reduced in elderly Alzheimer disease brain (UniProt) | |
| Isoform / epitope | No isoforms annotated; one 1–1081 chain (UniProt) |
Section 1
Recommended TSHZ3 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet) with published TSHZ3 staining methods for esophageal carcinoma and ureteropelvic junction tissue (PMC13265166; PMC5018636).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A08453); 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-TSHZ3, 2.5 μg/mL (datasheet A08453) |
| 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 | TSHZ3-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Ubiquitous 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 rule: nuclear antigen); compare citrate retrieval for carcinoma sections (PMC13265166).
Section 2
What Is the Expected TSHZ3 Staining Pattern?
TSHZ3 is expected mainly in nuclei: HPA describes ubiquitous nuclear tissue staining, while UniProt places the protein in the nucleus and reports no transmembrane segment (HPA: tissue IHC; UniProt Q63HK5: subcellular location, topology). HPA reports high staining in several epithelial, neural, hematopoietic, and adipose cell populations, but its tissue IHC reliability is Approved with low consistency between staining and RNA expression (HPA: tissue IHC).
| Distinct nuclear staining in the sampled cell population, with nuclei easier to identify against the counterstain. | This fits the reported tissue pattern and nuclear localization (HPA: ubiquitous nuclear expression; UniProt Q63HK5: nucleus). Judge intensity against the same run's controls; HPA's High labels describe particular cell populations, not a universal intensity threshold (HPA: tissue IHC). |
| Predominantly cytoplasmic, surface, or extracellular staining with little nuclear signal. | For tissue IHC, this departs from the expected dominant pattern and warrants an artefact or nonspecific staining check (HPA: ubiquitous nuclear expression). A membrane signal alone is not proof of error: HPA also reports plasma-membrane localization in ICC-IF, which is a different assay (HPA: subcellular ICC-IF). |
| Strong staining in a cell population expected to be low, or a pattern that tracks pigment or reaction deposits rather than nuclei. | Compare with morphology and controls before assigning TSHZ3: cross-reactivity or endogenous detection activity can produce misleading signal (general IHC practice). Liver cholangiocytes are reported Low, not absent, and HPA calls overall tissue staining ubiquitous (HPA: tissue IHC). |
| Haze across tissue, stroma, or empty spaces obscures nuclear boundaries. | Diffuse background limits interpretation even when some nuclei appear positive (general IHC practice). Review blocking, wash steps, detection chemistry, and exposure or development time before scoring; the supplied sources do not identify a TSHZ3-specific background mechanism (general IHC practice). |
| No convincing nuclear signal in a run that includes an HPA-reported High population. | First consider assay failure or poor control performance, then biological variation (general IHC practice). HPA reports High staining in cerebral-cortex glial cells and cerebellar granular-layer cells, among others, but its Approved tissue IHC has low staining–RNA consistency (HPA: tissue IHC). |
💡Expected TSHZ3 appearanceCall a positive IHC result when identifiable nuclei stain in the sampled cell population, especially one reported High by HPA; isolated diffuse or exclusively extranuclear colour should trigger control review (HPA: tissue IHC; general IHC practice).
| Which compartment anchors IHC scoring? | Nuclear staining anchors the call (HPA: ubiquitous nuclear tissue expression; UniProt Q63HK5: nucleus). UniProt also lists growth cones by similarity, so localization is context dependent (UniProt Q63HK5: subcellular location). |
| Which populations can serve as observed positives? | HPA reports High staining in adipocytes, bone-marrow hematopoietic cells, bronchial respiratory epithelial cells, and cerebral-cortex glial cells, among others (HPA: tissue IHC). These are observed patterns, not guaranteed assay controls (HPA: Approved; low staining–RNA consistency). |
| How strong is the tissue IHC evidence? | HPA labels tissue IHC Approved and notes low consistency with RNA expression; the listed HPA008834 antibody is IHC Approved, while HPA071010 has no listed IHC status (HPA: tissue IHC; HPA: antibody validation). Do not treat Approved as IHC Enhanced (HPA: antibody validation). |
| Do topology or processing predict a separate tissue compartment? | UniProt reports no transmembrane segment, signal peptide, propeptide, or annotated isoforms, and lists one 1–1081 chain (UniProt Q63HK5: topology, processing, isoforms). These annotations do not establish retrieval requirements or fixation sensitivity (UniProt Q63HK5: record scope). |
| What does IF/ICC add? | HPA reports enhanced nucleoplasmic localization and an approved additional plasma-membrane location in ICC-IF (HPA: subcellular ICC-IF). Use this as a localization cross-check; IF/ICC assay design belongs to its own guide (HPA: subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| No nuclear staining in an expected-positive area. | Detection, retrieval, or antibody conditions may have failed; an HPA High label is not a guarantee for every section (general IHC practice; HPA: tissue IHC). | Check the run's positive and detection controls, then verify retrieval and antibody conditions against the chosen assay instructions (general IHC practice). Compare with an HPA High population only after controls pass (HPA: tissue IHC). |
| Strong colour appears mainly outside nuclei. | The IHC pattern conflicts with HPA's ubiquitous nuclear description; staining chemistry or specificity may be contributing (HPA: tissue IHC; general IHC practice). | Review matched negative controls and cellular morphology, then repeat with adjusted detection conditions if controls implicate background (general IHC practice). Consider the separate ICC-IF membrane observation as context, not as IHC validation (HPA: subcellular ICC-IF). |
| Broad brown background obscures nuclei. | Nonspecific binding, incomplete washing, or excessive chromogen development can obscure chromogenic IHC (general IHC practice). | Check the negative control; review blocking, washing, and development time, then score only nuclei distinguishable from background (general IHC practice). |
| Signal follows blood-rich or pigmented areas rather than cell nuclei. | Endogenous detection activity or native pigment can mimic a chromogenic reaction (general IHC practice). | Compare controls and morphology; assess endogenous-enzyme blocking when the chosen detection chemistry requires it (general IHC practice). Do not count deposits without a cellular nuclear pattern (HPA: ubiquitous nuclear expression). |
| Liver cholangiocytes appear strongly positive. | This differs from HPA's Low cholangiocyte entry, although Low does not mean negative and HPA reports low staining–RNA consistency (HPA: tissue IHC). | Confirm cell identity and control behaviour, then compare nuclear signal with other populations in the same run (general IHC practice). Treat the discrepancy as a reason to investigate, not automatic proof of cross-reactivity (HPA: tissue IHC). |
| IF/ICC shows membrane signal alongside nuclear signal. | HPA lists an additional plasma-membrane location in ICC-IF, with nucleoplasm as the main location (HPA: subcellular ICC-IF). | Record both compartments and assess IF/ICC controls within that assay (general IF practice). Use the separate IF/ICC guide for its procedure; do not transfer its membrane result directly into chromogenic tissue IHC scoring (HPA: subcellular ICC-IF; HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing TSHZ3 staining
A08453 · IHCImmunohistochemistry of TSHZ3 in mouse brain tissue with TSHZ3 antibody at 2.5 μg/mL.
A08453 · IFImmunofluorescence of TSHZ3 in mouse brain tissue with TSHZ3 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000121297-TSHZ3/tissue/cerebral+cortex#imid_2663092
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000121297-TSHZ3/tissue/colon#imid_2663002
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000121297-TSHZ3/tissue/liver#imid_2663011
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000121297-TSHZ3/tissue/kidney#imid_2663004
HPA · ICC-IF ASC52teloHuman Protein Atlas immunofluorescence, ASC52telo, antibody HPA071010; green = TSHZ3, blue = nucleus, red = microtubules.
HPA · ICC-IF ASC52teloHuman Protein Atlas immunofluorescence, ASC52telo, antibody HPA071010; green = TSHZ3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TSHZ3 IHC & IF
🧪Run caudate first and expect staining in glial cells (HPA: High in caudate glial cells). HPA detects TSHZ3 in all 45 scored tissues and lists no negative tissue; use no-primary and isotype controls for the negative comparison, and treat cells without specific nuclear staining on the positive slide as a background reference rather than verified TSHZ3-negative cells (HPA: no negative rows; HPA: nucleoplasm enhanced).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: TSHZ3 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 TSHZ3 in ASC52telo, Rh30, U2OS, A-431, U-251MG, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) section, an isotype control matched to the primary antibody’s host and immunoglobulin class, and, if available, TSHZ3-knockout tissue or a cognate-peptide blocking control (standard IHC practice). For chromogenic caudate IHC, quench endogenous peroxidase and inspect tissue pigment before scoring (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported; the selected A08453 mouse-brain IHC caption gives 2.5 µg/mL but does not state a fixative (A08453 caption). Retrieval dependency and whether frozen sections or IF are easier are unreported in the supplied evidence. For caudate IF, assess age-related lipofuscin autofluorescence before assigning signal to TSHZ3 (standard IF practice).
HPA tissue IHC evidence for TSHZ3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: TSHZ3 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced TSHZ3 IHC Tips
Use nuclear staining as the primary IHC readout for TSHZ3, and interpret tissue patterns alongside the reported limits of antibody evidence (UniProt Q63HK5; HPA tissue IHC).
What retrieval should I start with for TSHZ3 in paraffin sections?
Start with Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval setting). Allow sections to cool in the retrieval buffer, then compare nuclear signal and tissue preservation against an otherwise identical section processed without retrieval (standard IHC practice). The main expected readout is nuclear because TSHZ3 is reported in the nucleus and HPA describes ubiquitous nuclear tissue staining (UniProt Q63HK5; HPA tissue IHC). If staining remains weak, test a milder citrate buffer at pH 6.0 as a fallback, keeping antibody concentration and detection constant (standard IHC practice).
How should I assess fixation effects on TSHZ3 staining?
Target-specific fixation sensitivity is unknown: the mouse brain IHC caption gives an antibody concentration of 2.5 µg/mL but does not state a fixative (caption A08453). Record the fixative, fixation duration, section age, and retrieval conditions for each specimen before comparing nuclear staining across samples (standard IHC practice). For a fixation comparison, process matched sections through the same Tris-EDTA pH 9.0, 95–98 °C, 20 min retrieval and chromogenic detection workflow (page retrieval setting; standard IHC practice). Judge each condition by nuclear signal, preserved morphology, and background, without assigning a TSHZ3-specific fixation effect from tissue staining patterns or sequence features (standard IHC practice).
Should cytoplasmic or membrane staining count as a TSHZ3-positive IHC result?
Prioritize nuclear staining when scoring paraffin sections: UniProt places TSHZ3 in the nucleus, and HPA describes ubiquitous nuclear tissue expression (UniProt Q63HK5; HPA tissue IHC). HPA also reports nucleoplasmic enrichment and an additional plasma membrane location in subcellular imaging, while UniProt reports growth-cone localization by similarity (HPA subcellular; UniProt Q63HK5). Because UniProt annotates no transmembrane segment, treat isolated membrane or diffuse cytoplasmic chromogen as a finding requiring confirmation, rather than scoring it automatically as positive (UniProt Q63HK5; standard IHC practice). Compare those patterns with nuclear signal on serial sections under the same pH 9.0, 20 min retrieval condition (page retrieval setting; standard IHC practice).
How can epitope uncertainty affect a negative TSHZ3 IHC result?
The record lists 0 annotated isoforms, but supplies no antibody epitope, so a negative section cannot be assigned to isoform exclusion or epitope loss (UniProt Q63HK5; supplied antibody evidence). TSHZ3 is a 1081-residue protein with an annotated phosphoserine at residue 682; whether that modification affects this antibody is unknown (UniProt Q63HK5; supplied antibody evidence). First compare nuclear staining across sections retrieved in Tris-EDTA pH 9.0 at 95–98 °C for 20 min, holding detection constant (page retrieval setting; standard IHC practice). If signal is absent, document epitope uncertainty and seek an independently characterized antibody before concluding that the specimen lacks TSHZ3 (standard IHC practice).
How should I cross-check the IHC pattern by multiplex IF?
Use IF as a cross-check of the nuclear chromogenic readout, with a validated marker for the cell population being examined and a nuclear counterstain; HPA reports high staining in cortical and caudate glial cells (HPA tissue IHC; standard IF practice). Choose a far-red TSHZ3 channel when specimen autofluorescence compromises shorter wavelengths, and include single-channel controls to assess bleed-through (standard IF practice). For an intracellular nuclear epitope, compare permeabilized and nonpermeabilized sections; a brief 0.1% Triton X-100 treatment is a general IF starting condition, while the antibody epitope is unreported (UniProt Q63HK5; standard IF practice; supplied antibody evidence). Score nuclear overlap with the cell marker and counterstain before relating IF to IHC (standard IF practice).
What should I check when TSHZ3 chromogenic staining is widespread?
Widespread nuclear staining alone is not evidence of background because HPA describes ubiquitous nuclear expression, although its tissue IHC reliability note reports low consistency with RNA expression (HPA tissue IHC). Inspect a no-primary control for detection background, block endogenous peroxidase for chromogenic detection, and check whether pigment or precipitate persists outside intact nuclei (standard IHC practice). Compare antibody titrations around the captioned 2.5 µg/mL only as an optimization exercise; that concentration comes from a mouse brain image, not a universal working dilution (caption A08453; standard IHC practice). Keep Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min fixed while changing antibody concentration (page retrieval setting; standard IHC practice).
How should I quantify TSHZ3 nuclear staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and region before analysis, then report the percentage of positive nuclei and an intensity-weighted nuclear H-score on a 0–300 scale (standard IHC practice). Normalize positive counts to the total evaluable nuclei in that same region; if reporting density, use positive nuclei per mm² of evaluable tissue (standard IHC practice). Keep section thickness, retrieval at Tris-EDTA pH 9.0 for 20 min, chromogen development, and image thresholds consistent across samples (page retrieval setting; standard IHC practice). Report cell populations separately where possible because HPA lists high staining in several distinct cell types and notes low staining consistency with RNA data (HPA tissue IHC).
How can I distinguish true TSHZ3 staining from artefact?
Give the strongest weight to staining within intact nuclei in an anatomically plausible cell population; HPA reports high staining in cortical glial cells and cerebellar granular-layer cells, while UniProt places TSHZ3 in the nucleus (HPA tissue IHC; UniProt Q63HK5). Scrutinize isolated membrane or diffuse cytoplasmic staining because HPA also reports an additional plasma membrane location, and UniProt annotates no transmembrane segment (HPA subcellular; UniProt Q63HK5). Exclude section edges, necrotic areas, pigment, and signal retained in a no-primary control when judging chromogen deposits (standard IHC practice). Interpret a weak or negative result alongside matched pH 9.0, 20 min retrieval controls and HPA's reported low staining-to-RNA consistency (page retrieval setting; HPA tissue IHC; standard IHC practice).
Boster reagents
Best TSHZ3 / Teashirt homolog 3 IHC Antibodies
A08453 has IHC and IF images from mouse brain tissue (catalog image captions) and lists Human, Mouse and Rat reactivity (catalog: reactivity).
Real IHC data
Anti-Teashirt homolog 3 TSHZ3 Antibody
Cat # A08453
A08453 is listed for IHC-P and IF (catalog: applications), with a mouse brain tissue IHC image at 2.5 μg/mL (catalog: IHC image caption). Its IF image also shows mouse brain tissue (catalog: IF image caption); Human, Mouse and Rat are listed as reactive species (catalog: reactivity).
Which to pick: Choose A08453 for paraffin-section IHC because IHC-P is listed (catalog: applications); its IHC image shows mouse brain tissue, but the fixative is unreported (catalog: IHC image caption). For IF, A08453 has a mouse brain tissue image (catalog: IF image caption); ICC validation and clonality are unreported (catalog: applications and clone). For Human or Rat samples, A08453 lists reactivity (catalog: reactivity), while the pictured IHC and IF data are from mouse tissue (catalog image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q63HK5 (TSH3_HUMAN, Teashirt homolog 3).
- Human Protein Atlas. TSHZ3 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. TSHZ3 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the plasma membrane..
- Human Protein Atlas. TSHZ3 antibody validation summary (2 antibodies).
- The transcription factor TSHZ3 promotes tumor immunosuppression and inhibits metastasis in lung adenocarcinoma. Frontiers in immunology 2025 — PMC12011852.
- Prognostic value and potential upstream regulator of Schwann cells in esophageal squamous cell carcinoma. Translational cancer research 2026 — PMC13265166.
- Sonic hedgehog, TBX18, and TSHZ3 proteins involved in pyeloureteral motility development are overexpressed in ureteropelvic junction obstruction. An immunohistochemical, histopathological, and clinical comparative study. Saudi medical journal 2016 — PMC5018636.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:10819331 — UniProt-cited evidence.
- PubMed PMID:11230166 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TSHZ3 (Teashirt homolog 3). Expected localisation Ubiquitous nuclear tissue staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A08453 at 2.5 μg/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 workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A08453); 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-TSHZ3, 2.5 μg/mL (datasheet A08453); 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 A08453); Caveat: Staining has low concordance with RNA expression (HPA tissue IHC); Isoform / epitope: No isoforms annotated; one 1–1081 chain (UniProt); Regulation: Reduced in elderly Alzheimer disease brain (UniProt). Evidence: UniProt Q63HK5, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A08453, PMC13265166, PMC5018636. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TSHZ3/immunohistochemistry-ihc-guide.