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TSHZ2 / Teashirt homolog 2 · IHC design guide
Design Immunohistochemistry for TSHZ2
Plan paraffin-section TSHZ2 IHC with catalog antibody A08863 at 1:100–1:300 and HPA-supported positive tissue controls (datasheet; HPA tissue IHC). Score nuclear and cytoplasmic staining while accounting for the uncertain tissue-IHC reliability (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TSHZ2 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 | Nuclear (UniProt); nuclear and cytoplasmic in tissue (HPA tissue IHC) | |
| Staining pattern | Widespread nuclear and cytoplasmic tissue staining (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Liver |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Tissue staining has low concordance with RNA expression (HPA tissue IHC) | |
| Regulation | Reduced in elderly Alzheimer brains (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unreported (UniProt) |
Section 1
Recommended TSHZ2 IHC & IF Protocols
Compare the catalog antibody IHC-P protocol with the published lung adenocarcinoma tissue protocol (PMC8040402).
| Sample | Paraffin-embedded human prostate carcinoma tissue; fixative not specified (datasheet A08863) |
| Fixation | Image fixative and duration unreported (datasheet A08863); 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-TSHZ2, 1:100 - 1:300 (datasheet A08863) |
| 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 | TSHZ2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear and cytoplasmic expression in all tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page antigen retrieval); citrate buffer was used in the published protocol (PMC8040402).
Section 2
What Is the Expected TSHZ2 Staining Pattern?
TSHZ2 is annotated in the nucleus and has no transmembrane segment (UniProt Q9NRE2). In paraffin tissue sections, HPA reports nuclear and cytoplasmic staining across tissues, including High staining in cerebral cortex neurons, adrenal glandular cells and placental decidual cells (HPA tissue IHC). Treat that broad pattern cautiously: HPA rates its tissue IHC reliability Uncertain because antibody staining and RNA expression have low consistency (HPA tissue IHC).
| Neuronal nuclei stain in cerebral cortex; some cytoplasmic signal is also present (HPA tissue IHC). | This fits the reported High neuronal staining and broad nuclear and cytoplasmic tissue pattern (HPA tissue IHC). Nuclear signal also agrees with the UniProt localization (UniProt Q9NRE2). Record nuclear and cytoplasmic scores separately; HPA's Uncertain reliability limits how confidently either signal can be assigned to TSHZ2 (HPA tissue IHC). |
| Signal is confined to cell borders, lumens or extracellular material, with no convincing cellular staining. | This does not match UniProt's nuclear localization or HPA's reported nuclear and cytoplasmic tissue pattern (UniProt Q9NRE2; HPA tissue IHC). Inspect morphology and the negative detection control before calling it TSHZ2; misplaced chromogen or detection activity is a plausible artefact (general IHC practice). |
| Cholangiocytes stain strongly while expected positive cells on the same run stain weakly or not at all. | HPA reports TSHZ2 as Not detected in liver cholangiocytes, although its tissue IHC reliability is Uncertain (HPA tissue IHC). Check for cross-reactivity or endogenous detection activity using appropriate controls (general IHC practice). A cholangiocyte signal alone cannot establish a new TSHZ2 expression pattern. |
| Brown signal spreads across stroma, section edges or blank areas and obscures cell boundaries. | Diffuse deposition is difficult to score as cellular TSHZ2; HPA's reported pattern is nuclear and cytoplasmic (HPA tissue IHC). Compare the negative detection control, then review blocking, washing and chromogen development (general IHC practice). Do not interpret background intensity as a tissue staining level. |
| No staining appears in cerebral cortex neurons or adrenal glandular cells. | Both populations are reported High by HPA tissue IHC, making them useful run controls, though HPA rates the tissue result Uncertain (HPA tissue IHC). Check section integrity, antigen retrieval, antibody dilution and detection controls before treating a blank section as biological absence (general IHC practice). |
💡Expected TSHZ2 appearanceA plausible positive IHC result shows cellular nuclear staining, potentially with cytoplasmic staining, in High HPA populations such as cerebral cortex neurons; strong cell-border-only or extracellular deposits warrant artefact checks (UniProt Q9NRE2; HPA tissue IHC; general IHC practice).
| Tissue IHC reliability | HPA calls the pattern Uncertain because staining and RNA expression have low consistency (HPA tissue IHC). Use reported levels as comparisons, not definitive identity tests. |
| Compartment evidence differs by assay | UniProt places TSHZ2 in the nucleus; HPA tissue IHC reports nuclear and cytoplasmic staining, while HPA ICC-IF approves Golgi and nucleoplasmic localization (UniProt Q9NRE2; HPA tissue IHC; HPA subcellular). The ICC-IF result is context for interpretation, not a paraffin IHC protocol. |
| Antibody validation | HPA038123 is Uncertain for IHC and Approved for ICC (HPA antibodies). Its ICC status does not resolve uncertainty in a paraffin section; compare IHC controls and morphology (HPA antibodies; general IHC practice). |
| Isoforms and topology | UniProt lists 2 isoforms, no signal peptide and no transmembrane segment (UniProt Q9NRE2). The supplied record gives no antibody epitope, so isoform coverage cannot be inferred. |
| Fixation sensitivity | Target-specific fixation sensitivity is unreported in the supplied UniProt and HPA records. Document fixation and retrieval conditions for comparison between IHC runs (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in the expected positive control. | A failed staining step or unsuitable run conditions are possible (general IHC practice); HPA reports High staining in cerebral cortex neurons (HPA tissue IHC). | Verify tissue identity and section quality, then review retrieval, the IHC-validated antibody dilution and detection reagents (general IHC practice). |
| Strong signal appears in liver cholangiocytes. | HPA lists cholangiocytes as Not detected, but the tissue IHC pattern is Uncertain (HPA tissue IHC). Cross-reactivity or endogenous detection activity is possible (general IHC practice). | Compare an appropriate negative detection control and a positive tissue in the same run; report the discordance rather than assigning specificity from intensity alone (general IHC practice). |
| Only sharp cell-border staining is visible. | That distribution conflicts with the nuclear UniProt annotation and HPA's nuclear and cytoplasmic tissue pattern (UniProt Q9NRE2; HPA tissue IHC). | Check whether signal follows tissue edges or deposits, and compare the negative detection control before scoring cells (general IHC practice). |
| Diffuse brown background obscures nuclei. | Nonspecific deposition or excess detection signal can reduce contrast (general IHC practice). | Review blocking, washing, antibody dilution and chromogen development; score only identifiable cellular signal (general IHC practice). |
| Cytoplasmic signal exceeds nuclear signal. | HPA reports cytoplasmic tissue staining, while UniProt annotates a nuclear location and HPA rates tissue IHC Uncertain (HPA tissue IHC; UniProt Q9NRE2). | Score compartments separately and compare positive and negative controls. Do not equate cytoplasmic intensity alone with confirmed TSHZ2 localization. |
| The ICC-IF Golgi pattern is absent from a paraffin IHC section. | HPA's approved Golgi localization comes from ICC-IF; its tissue IHC summary describes nuclear and cytoplasmic staining (HPA subcellular; HPA tissue IHC). | Interpret the paraffin section against the tissue IHC pattern and controls. Use the separate IF/ICC guide for fluorescence-specific decisions. |
Experiment evidence
IHC & IF images showing TSHZ2 staining
A08863 · IHCImmunohistochemistry analysis of paraffin-embedded human prostate carcinoma tissue, using TSH2 Antibody. The picture on the right is blocked
A08863 · IFImmunofluorescence analysis of MCF7 cells, using TSH2 Antibody. The picture on the right is blocked with the synthesized peptide.
A08863-1 · IFImmunofluorescence of TSHZ2 in A20 cells with TSHZ2 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000182463-TSHZ2/tissue/cerebral+cortex#imid_9456548
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000182463-TSHZ2/tissue/colon#imid_9456482
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000182463-TSHZ2/tissue/liver#imid_9456475
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000182463-TSHZ2/tissue/kidney#imid_9456468
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA038123; green = TSHZ2, blue = nucleus, red = microtubules.
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA038123; green = TSHZ2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TSHZ2 IHC & IF
🧪Run cerebral cortex first: neuronal cells should stain (High; HPA: Cerebral cortex, neuronal cells). Use liver cholangiocytes as the negative tissue (HPA: Liver, cholangiocytes, Not detected); any independently verified negative cells within the cortex section should show only background chromogen, since the supplied HPA rows do not identify a negative cell type there.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Liver (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TSHZ2 in HeLa, SiHa, U2OS, with annotated localisation: Golgi apparatus (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and clonality, and a target knockout control if available; peptide blocking is also documented for the catalog antibody (selected A08863 tissue-IHC caption: prostate carcinoma section blocked with synthesized peptide). Quench endogenous peroxidase for chromogenic detection and distinguish neuronal pigment from specific staining; for IF, assess tissue autofluorescence with a no-primary control (standard IHC/IF practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the fixative in the selected A08863 paraffin-section caption is unreported (selected A08863 tissue-IHC caption). Antigen retrieval may need empirical optimization for paraffin IHC; the supplied evidence does not establish a TSHZ2-specific retrieval dependency or show that frozen sections or IF are easier (standard IHC practice). In cerebral cortex, neuronal pigment can complicate chromogenic scoring, while autofluorescence can complicate IF (standard IHC/IF practice).
HPA tissue IHC evidence for TSHZ2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Placenta | Decidual cells | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TSHZ2 IHC Tips
Use compartment-aware controls when optimizing chromogenic TSHZ2 IHC: UniProt and HPA report different localisation patterns, and HPA rates tissue staining reliability as uncertain.
How should I retrieve TSHZ2 antigen in paraffin sections?
Start with Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval setting). Let sections cool in retrieval buffer before washing, and keep heating and cooling consistent across the comparison set (standard IHC practice). Assess nuclear and cytoplasmic or Golgi-region staining separately because the reported TSHZ2 locations differ between sources (UniProt Q9NRE2: nucleus; HPA subcellular: Golgi apparatus and nucleoplasm; HPA tissue IHC: nuclear and cytoplasmic). Include a matched section for peptide competition when using A08863, whose paraffin-section image includes that control (A08863 caption). If signal remains weak, trial citrate pH 6.0 on adjacent sections as a fallback and compare morphology and background (standard IHC optimization).
Does fixation change TSHZ2 staining in paraffin sections?
The A08863 image shows paraffin-embedded human prostate carcinoma with a peptide-blocked comparison, but its caption does not state the fixative (A08863 caption). TSHZ2-specific fixation sensitivity is therefore unknown; the HPA staining profile and UniProt annotations do not establish a fixation response (A08863 caption; HPA tissue IHC; UniProt Q9NRE2). For a prospective comparison, document the fixative and compare matched samples fixed for 12 versus 24 h, holding retrieval, antibody incubation and detection constant (standard IHC optimization). For archived sections with different fixation histories, interpret intensity within each processing group and inspect nuclear detail and tissue preservation before attributing a weak result to biology (standard IHC practice).
Should TSHZ2 staining be nuclear or cytoplasmic?
Score nuclear staining explicitly because UniProt places TSHZ2 in the nucleus and describes a probable transcriptional regulator (UniProt Q9NRE2). Also document cytoplasmic or perinuclear signal separately: HPA tissue IHC reports nuclear and cytoplasmic expression, while its subcellular data place TSHZ2 mainly at the Golgi apparatus with additional nucleoplasmic localisation (HPA tissue IHC; HPA subcellular). Do not merge these compartments into one positive score, particularly because HPA calls tissue-IHC reliability uncertain owing to low consistency with RNA expression (HPA tissue IHC). Compare the same cell class, compartment and staining pattern across serial sections, and use peptide competition for A08863 as an antibody-binding control (standard IHC scoring practice; A08863 caption).
Can the antibody distinguish TSHZ2 isoforms or modified epitopes?
Treat isoform recognition as unresolved until the antibody's immunogen and epitope map are available: UniProt lists 2 TSHZ2 isoforms, while the supplied A08863 caption gives neither map (UniProt Q9NRE2; A08863 caption). UniProt also annotates phosphoserine at residue 980, but this alone does not show whether phosphorylation changes staining (UniProt Q9NRE2). The peptide-blocked paraffin image supports competition with the supplied peptide in that image; it does not establish isoform discrimination or exclude every off-target interaction (A08863 caption; standard antibody-validation principle). If isoform attribution matters, obtain epitope coordinates and compare them with isoform sequences before interpreting compartment-specific differences (standard epitope-mapping practice).
How should I adapt TSHZ2 localisation checks for multiplex immunofluorescence?
Treat IF/ICC as a separate assay: HPA provides subcellular images from HeLa, SiHa and U2OS cells, while the supplied A08863 tissue caption documents paraffin IHC only (HPA subcellular; A08863 caption). For tissue multiplexing, pair TSHZ2 with a marker identifying the cell class being scored, such as a neuronal marker when examining cerebral cortex, where HPA reports high neuronal staining (HPA tissue IHC; standard multiplex IF practice). Choose a far-red TSHZ2 channel when shorter wavelengths show tissue autofluorescence, and check single-stain controls for bleed-through (standard IF practice). Because reported sites include nucleoplasm and Golgi and UniProt lists no transmembrane segment, test mild permeabilisation, such as 0.1% Triton X-100 for 5 min, while checking organelle morphology (HPA subcellular; UniProt Q9NRE2 topology; standard IF optimization).
What should I check when TSHZ2 chromogenic staining is diffuse?
First compare the stained section with a no-primary control and inspect edge staining, damaged areas and deposits before assigning diffuse colour to TSHZ2 (standard chromogenic IHC practice). For peroxidase detection, a 3% hydrogen peroxide block for about 10 min and controlled DAB development address endogenous enzyme activity and excessive chromogen deposition as general workflow steps (standard IHC practice). Titrate the IHC-validated antibody and blocking conditions on matched sections, keeping retrieval fixed at Tris-EDTA pH 9.0 during that comparison (standard IHC optimization; page retrieval setting). A08863 has a peptide-blocked paraffin image, which is useful for assessing peptide-competitive staining but is not a complete specificity test (A08863 caption; standard antibody-validation principle).
How should I quantify TSHZ2 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell class and compartment before scoring, then record nuclear and cytoplasmic or perinuclear staining separately (UniProt Q9NRE2: nucleus; HPA tissue IHC: nuclear and cytoplasmic; HPA subcellular: Golgi and nucleoplasm). For chromogenic sections, report the percentage of positive cells and an H-score from 0–300, calculated as each intensity category's cell percentage multiplied by its 0–3 grade (standard IHC scoring practice). Normalise cell counts to the number of evaluable cells in the specified class; if measuring positive-cell density, report cells per mm² of evaluable tissue (standard histological quantification). Apply one threshold, sampling plan and exclusion rule across slides, and interpret small intensity differences cautiously because HPA rates tissue staining reliability uncertain (standard IHC scoring practice; HPA tissue IHC).
How can I distinguish genuine TSHZ2 signal from artefact?
A credible result should have reproducible staining in intact cells, a defined compartment and lower signal in the matched peptide-blocked A08863 section (standard IHC interpretation; A08863 caption). Interpret nuclear staining alongside UniProt's nuclear annotation, but record Golgi-region or cytoplasmic staining separately because HPA reports those locations and calls tissue-IHC reliability uncertain (UniProt Q9NRE2; HPA subcellular; HPA tissue IHC). Check the identified cell class against the reported pattern, such as high neuronal staining in cerebral cortex or undetected cholangiocyte staining in liver, without treating those examples as absolute controls (HPA tissue IHC). Exclude edge effects, necrotic regions and colour persisting in a no-primary or peroxidase-control section before calling a specimen positive (standard chromogenic IHC practice).
Boster reagents
Best TSHZ2 / Teashirt homolog 2 IHC Antibodies
The catalog includes TSHZ2 antibodies with images from paraffin-section IHC and cell IF (catalog image captions); listed reactivity covers human and mouse for both SKUs, plus rat for A08863-1 (catalog reactivity).
Real IHC data
Anti-TSH2 TSHZ2 Antibody
Cat # A08863
Real IF data
Anti-Teashirt homolog 2 TSHZ2 Antibody
Cat # A08863-1
A08863 is listed for IHC and IF, with images of paraffin-embedded human prostate carcinoma tissue and MCF7 cells (catalog applications and image captions). A08863-1 is listed for IF, with an image of A20 cells at 20 μg/mL (catalog applications and IF image caption).
Which to pick: Choose the polyclonal A08863 for tissue IHC: its own image shows a paraffin-embedded human prostate carcinoma section, and its listed IHC dilution is 1:100–1:300 (A08863 catalog dilution and IHC image caption); the fixative is unreported (A08863 IHC image caption). For IF/ICC, A08863 lists both applications and shows MCF7 cell IF, while A08863-1 lists IF and shows A20 cell IF at 20 μg/mL but does not list ICC (catalog applications and respective IF image captions). For rat samples, A08863-1 is the SKU with listed rat reactivity, although its pictured IF sample is A20 cells; A08863 lists human and mouse reactivity (catalog reactivity and A08863-1 IF image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NRE2 (TSH2_HUMAN, Teashirt homolog 2).
- Human Protein Atlas. TSHZ2 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. TSHZ2 subcellular location (ICC-IF): Mainly localized to the Golgi apparatus. In addition localized to the nucleoplasm..
- Human Protein Atlas. TSHZ2 antibody validation summary (1 antibodies).
- Overexpression of Teashirt Homolog 2 suppresses cell proliferation and predicts the favorable survival of Lung Adenocarcinoma. International journal of medical sciences 2021 — PMC8040402.
- Down-regulation of the zinc-finger homeobox protein TSHZ2 releases GLI1 from the nuclear repressor complex to restore its transcriptional activity during mammary tumorigenesis. Oncotarget 2016 — PMC4868714.
- Perturbed cell fate decision by schizophrenia-associated AS3MT(d2d3) isoform during corticogenesis. Science advances 2025 — PMC11952104.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:11780052 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TSHZ2 (Teashirt homolog 2). Expected localisation Nuclear (UniProt); nuclear and cytoplasmic in tissue (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A08863 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Liver. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human prostate carcinoma tissue; fixative not specified (datasheet A08863); 2. Fixation: Image fixative and duration unreported (datasheet A08863); 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-TSHZ2, 1:100 - 1:300 (datasheet A08863); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Tissue staining has low concordance with RNA expression (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unreported (UniProt); Regulation: Reduced in elderly Alzheimer brains (UniProt). Evidence: UniProt Q9NRE2, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the Golgi apparatus. In addition localized to…, Datasheet A08863, PMC8040402. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TSHZ2/immunohistochemistry-ihc-guide.