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GATA3 / Trans-acting T-cell-specific transcription factor GATA-3 · IHC design guide
Design Immunohistochemistry for GATA3
Plan chromogenic GATA3 IHC around nuclear staining in selected tissues (HPA tissue IHC). Compare tissue controls using the catalog antibody's 0.5–1 μg/mL IHC-P range (datasheet A00593-1), and assess possible off-target staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GATA3 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 staining in selected tissues (HPA tissue IHC) | |
| Staining pattern | Parathyroid glandular and skin basal cells: nuclear staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A00593-1) | |
| Positive control | Kidney+4 more · see all | |
| Negative control | Adipose tissue+4 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 A00593-1) | |
| Caveat | Presumed off-target staining can complicate interpretation (HPA tissue IHC) | |
| Regulation | IL33-linked macrophage response (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended GATA3 IHC & IF Protocols
The catalog antibody's IHC-P protocol (datasheet A00593-1) and one published FFPE tumor protocol (PMC10969368) provide starting conditions for GATA3 staining.
| Sample | Paraffin-embedded human mammary cancer tissue; fixative not specified (datasheet A00593-1) |
| Fixation | Image fixative and duration unreported (datasheet A00593-1); 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 A00593-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A00593-1) |
| Primary antibody | Rabbit anti-GATA3, 0.5-1μg/ml (datasheet A00593-1) |
| Primary incubation | Overnight at 4 °C (datasheet A00593-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A00593-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GATA3-positive staining in cells in glomeruli of kidney (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in selected tissues, including parathyroid gland and skin. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A00593-1); assess nuclear staining with appropriate controls (UniProt P23771: nucleus).
Section 2
What Is the Expected GATA3 Staining Pattern?
GATA3 is a nuclear transcription factor with no transmembrane segment (UniProt P23771: nucleus; topology). In paraffin IHC, expect nuclear staining in selected cells: high in kidney glomerular cells and medium in breast and parathyroid glandular cells, among other reported sites (HPA tissue IHC). HPA rates the tissue profile Enhanced, while noting medium consistency with RNA data and presumed off-target staining that was disregarded (HPA tissue IHC: reliability).
| Distinct nuclear staining in cells of the kidney glomeruli, with little staining outside their nuclei (HPA tissue IHC: High in glomerular cells; UniProt P23771: nucleus). | This fits the reported high-signal tissue pattern and the expected compartment. Score the stained cell population and nuclear signal separately; a strong field-wide color reaction alone does not establish GATA3 positivity (HPA tissue IHC; standard IHC practice). |
| Predominantly cytoplasmic or membrane staining, with nuclei remaining clear (UniProt P23771: nucleus; HPA subcellular: supported nucleoplasm). | This conflicts with the supported localization. Treat it as a staining artifact or nonspecific signal until a nuclear pattern can be demonstrated with suitable controls; GATA3 has no annotated transmembrane segment (UniProt P23771: topology; standard IHC practice). |
| Prominent staining in cells reported as undetected, such as adipocytes or adrenal glandular cells (HPA tissue IHC: Not detected in these cell types). | Question antibody specificity or endogenous detection activity before interpreting those cells as positive. HPA’s negative calls apply to the named cell populations, not every cell in the tissue; its profile also notes presumed off-target binding (HPA tissue IHC: reliability; standard IHC practice). |
| A diffuse brown haze spreads across tissue, including spaces between cells (standard IHC practice). | Diffuse background does not reproduce the selected nuclear pattern (HPA tissue IHC: nuclear expression). Review blocking, washing, chromogen development and endogenous enzyme controls for the detection system used (standard IHC practice). |
| No nuclear signal appears in kidney glomerular cells on the same run (HPA tissue IHC: High in glomerular cells). | The run may be technically insensitive; tissue condition or the actual sampled cell population may also matter (standard IHC practice). Check that glomerular cells are present, then review the antibody’s validated IHC-P conditions and detection controls before calling the sample negative (HPA tissue IHC; standard IHC practice). |
💡Expected GATA3 appearanceCall a result positive when staining is chiefly nuclear in the relevant cells, especially strong in kidney glomerular cells or moderate in reported glandular populations; diffuse cytoplasmic color or widespread staining of reported-negative cells is suspect (HPA tissue IHC; UniProt P23771: nucleus).
| Cell population and tissue choice (HPA tissue IHC) | Kidney glomerular cells are reported High; breast, epididymis, parathyroid and rectal glandular cells are Medium, as are placental syncytiotrophoblasts and skin basal-layer cells. Compare the named cells rather than assigning one expected intensity to a whole section (HPA tissue IHC). |
| Limits of a negative comparator (HPA tissue IHC) | Adipocytes, adrenal and appendix glandular cells, bone marrow hematopoietic cells and bronchial respiratory epithelial cells are reported Not detected. That wording describes those cells in the HPA observations; it does not establish a universal absence of GATA3 in every specimen or condition (HPA tissue IHC). |
| Evidence strength and antibody selection (HPA tissue IHC; HPA antibodies) | The tissue profile is Enhanced but has medium staining-to-RNA consistency, with presumed off-target binding disregarded. CAB016217 has Enhanced IHC validation; HPA029730 has Supported ICC validation and no supplied IHC rating. Keep each validation attached to its stated application (HPA tissue IHC; HPA antibodies). |
| What does IF/ICC add? (HPA subcellular) | HPA reports supported nucleoplasmic localization in ICC-IF, consistent with a nuclear IHC readout (HPA subcellular; UniProt P23771: nucleus). It supports the compartment expectation, but its images and ICC validation do not supply an IHC-P staining condition (HPA subcellular; HPA antibodies). |
| Isoforms and protein processing (UniProt P23771) | UniProt lists two isoforms and a full-length 1–443 chain, with no signal peptide or propeptide annotated. The supplied record does not map an antibody epitope to either isoform, so it cannot justify an isoform-specific tissue interpretation (UniProt P23771: isoforms; processing). |
| Situation | Likely cause | Next action |
|---|---|---|
| Kidney glomerular cells lack the expected nuclear color (HPA tissue IHC: High). | The sampled section may lack the named cells, or the run may have weak detection (standard IHC practice). | Confirm glomerular cells on the section and assess a known-positive control; then check the catalog antibody’s validated IHC-P instructions and detection steps (HPA tissue IHC; standard IHC practice). |
| Brown signal outlines cells or fills cytoplasm while nuclei stay pale (UniProt P23771: nucleus). | The distribution is inconsistent with the supported nuclear and nucleoplasmic localization (UniProt P23771; HPA subcellular). | Review morphology, counterstain and detection controls; require reproducible nuclear signal before scoring GATA3 positivity (standard IHC practice). |
| Reported-negative cells stain strongly, including adipocytes (HPA tissue IHC: Not detected in adipocytes). | Nonspecific binding or endogenous detection activity is plausible; HPA notes presumed off-target staining in its tissue assessment (HPA tissue IHC: reliability; standard IHC practice). | Compare a reagent-omission or detection-only control and a named positive population, then reassess specificity in the same run (standard IHC practice; HPA tissue IHC). |
| Most of the section has diffuse chromogen deposit (standard IHC practice). | Background from blocking, washing, detection chemistry or chromogen development can obscure the expected nuclear pattern (standard IHC practice; HPA tissue IHC). | Check the applicable blocking and wash steps, endogenous enzyme control and chromogen development time for the detection system (standard IHC practice). |
| A breast section stains less strongly than a kidney section (HPA tissue IHC: Medium in breast glandular cells; High in glomerular cells). | The reported levels differ by named cell population, so equal intensity is not the expected comparison (HPA tissue IHC). | Judge breast glandular nuclei against the reported medium pattern and the run’s controls; avoid using kidney intensity as a required threshold (HPA tissue IHC; standard IHC practice). |
| ICC-IF shows a nucleoplasmic pattern, but IHC-P gives a different result (HPA subcellular: supported nucleoplasm). | The supplied HPA antibody ratings are application-specific: Supported ICC for HPA029730 and Enhanced IHC for CAB016217 (HPA antibodies). | Interpret the IHC-P section with its own validated antibody and tissue controls; use ICC-IF only to inform the expected compartment (HPA antibodies; HPA subcellular; standard IHC practice). |
Experiment evidence
IHC & IF images showing GATA3 staining
A00593-1 · IHCIHC analysis of GATA3 using anti-GATA3 antibody (A00593-1). GATA3 was detected in paraffin-embedded section of human mammary cancer tissue.
A00593-1 · IFIF analysis of GATA3 using anti-GATA3 antibody (A00593-1). GATA3 was detected in immunocytochemical section of Hela cells. Enzyme antigen re
M00593 · IHCImmunohistochemical analysis of paraffin-embedded Rat intestine, using the Antibody at 1:500 dilution.
M00593 · IHCImmunohistochemical analysis of paraffin-embedded Human pituitary tumor, using the Antibody at 1:250 dilution.
A00593 · IHCImmunohistochemistry of GATA3 in rat brain tissue with GATA3 antibody at 2.5 μg/mL.
A00593 · IHCImmunohistochemistry of GATA3 in mouse brain tissue with GATA3 antibody at 2.5 μg/ml.
A00593 · IFImmunofluorescence of GATA3 in Rat Brain tissue with GATA3 antibody at 20 μg/mL.
P00593 · IHCImmunohistochemical analysis of paraffin-embedded human bladder cancer, using Phospho-GATA3 (S308) Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000107485-GATA3/tissue/cerebral+cortex#imid_17358383
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000107485-GATA3/tissue/colon#imid_17358317
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000107485-GATA3/tissue/liver#imid_17358310
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000107485-GATA3/tissue/kidney#imid_17358311
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029730; green = GATA3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029730; green = GATA3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GATA3 IHC & IF
🧪Run kidney first: cells in glomeruli should show nuclear staining (HPA: Kidney, cells in glomeruli High; UniProt P23771: Nucleus). Use adipose tissue adipocytes as a negative tissue (HPA: Adipose tissue, adipocytes Not detected); on the kidney slide, treat cells outside the stained glomerular population that lack nuclear signal as internal negative comparators, without assuming every other renal cell is negative (HPA: Kidney, cells in glomeruli High).
Positive control tissue: Kidney (Cells in glomeruli, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GATA3 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only), a rabbit isotype control matched to the primary where applicable, and validated GATA3 knockout material or a peptide-block control if the immunizing peptide is available (selected A00593-1 caption: rabbit primary; standard IHC practice). Block endogenous peroxidase and assess endogenous biotin in kidney sections when using the caption’s biotin–SABC/DAB detection method (selected A00593-1 caption: biotinylated secondary, SABC and DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the fixative for the selected A00593-1 paraffin-section caption is unreported (selected A00593-1 caption). That caption uses heat retrieval in EDTA at pH 8.0, but provides no comparison establishing retrieval dependency (selected A00593-1 caption). The supplied evidence does not establish whether frozen sections or IF are easier; renal endogenous biotin can complicate the caption’s biotin-based detection (HPA: supported nucleoplasmic ICC-IF localisation; selected A00593-1 caption: SABC detection; standard IHC practice).
HPA tissue IHC evidence for GATA3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). 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 |
|---|---|---|---|---|
| Kidney | Cells in glomeruli | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Medium | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Placenta | Syncytiotrophoblasts | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GATA3 IHC Tips
Use nuclear staining and matched tissue controls to troubleshoot GATA3 in paraffin sections; the selected antibody has a documented chromogenic IHC example (datasheet A00593-1).
How should I adjust retrieval when GATA3 nuclear staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A00593-1). The documented example then used 1 μg/ml primary antibody overnight at 4°C, so check whether those conditions were reproduced before changing retrieval (datasheet A00593-1). Compare a retrieval-treated control section with a matched section processed without retrieval, keeping detection and DAB development consistent (standard IHC practice). If staining remains weak, adjust heating or cooling conditions one variable at a time and assess whether nuclear signal improves without raising background (standard IHC practice; UniProt P23771: nuclear localisation). Record the retrieval conditions used for every comparison (standard IHC practice).
Could fixation explain absent GATA3 staining in these paraffin sections?
The selected tissue-IHC caption identifies a paraffin section but does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet A00593-1). Record the actual fixative and processing history for each specimen before attributing absent nuclear signal to fixation (standard IHC practice; UniProt P23771: nuclear localisation). Compare specimens with similar processing and include a section with documented GATA3 nuclear staining as a positive run control (standard IHC practice; datasheet A00593-1). If the control stains while one specimen does not, evaluate tissue preservation and staining distribution before changing the antibody conditions (standard IHC practice). Do not infer fixation tolerance from tissue-expression patterns (HPA: tissue IHC profile).
Should diffuse cytoplasmic staining be counted as GATA3 positive?
Score nuclear staining as the expected GATA3 signal in chromogenic IHC (UniProt P23771: nucleus; HPA: nuclear expression in selected tissues). The supported subcellular location is nucleoplasm, and the protein has no annotated transmembrane segment (HPA: supported nucleoplasm; UniProt P23771: topology). Inspect nuclei at high magnification and compare their DAB signal with nearby unstained nuclei and the counterstain (standard IHC practice). Treat diffuse cytoplasmic colour without convincing nuclear staining as questionable, then review the no-primary control, DAB development and section quality (standard IHC practice). Document any mixed pattern separately from the nuclear score so that cytoplasmic background does not inflate the result (standard IHC practice).
Can isoforms or phosphorylation explain discordant GATA3 staining?
GATA3 has 2 annotated isoforms, but the supplied product caption does not identify the antibody epitope or establish isoform coverage (UniProt P23771: isoforms 1 and 2; datasheet A00593-1). Phosphoserine residues are annotated at positions 115 and 162; their effect on this antibody's tissue staining is unknown (UniProt P23771: modified residues; datasheet A00593-1). Check whether an available epitope specification maps to sequence shared by both isoforms before interpreting a negative section as absence of all GATA3 (standard IHC practice). Compare the staining with a positive control processed in the same run, and keep any isoform or phosphorylation explanation provisional without direct validation (standard IHC practice).
How can I investigate a disputed nuclear IHC pattern by IF?
Use IF as an independent localisation check and report it separately from the chromogenic IHC result (standard IHC/IF practice). Multiplex GATA3 with a marker of an expected cell population, such as CD3 for T cells, and assess whether their nuclei contain the GATA3 signal (UniProt P23771: T-cell expression and nuclear localisation; standard IF practice). Choose fluorophores after checking tissue autofluorescence in an unstained section, and include single-channel controls to assess bleed-through (standard IF practice). Because GATA3 is nuclear and has no transmembrane segment, use a permeabilisation step compatible with nuclear antibody access, then verify nuclear boundaries with a counterstain (UniProt P23771: localisation and topology; standard IF practice).
What should I check when DAB appears throughout the section?
The selected example used 10% goat serum blocking, a biotinylated secondary antibody and streptavidin–biotin detection with DAB (datasheet A00593-1). Compare the stained slide with no-primary and detection-only controls to locate background introduced by the secondary or detection steps (standard IHC practice). Check endogenous peroxidase blocking, nonspecific antibody binding and endogenous biotin when troubleshooting this chromogenic workflow (standard IHC practice; datasheet A00593-1: biotin-based detection). Reduce excessive primary incubation or DAB development only after confirming that a positive control retains distinct nuclear signal (standard IHC practice; UniProt P23771: nuclear localisation). Record the distribution of background, including edges and damaged tissue (standard IHC practice).
How should I score heterogeneous GATA3 nuclear staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and regions to score before examining the chromogenic slides, then exclude damaged areas consistently (standard IHC practice). Record the percentage of cells with nuclear DAB staining and, when intensity matters, calculate an H-score from intensity categories and their corresponding percentages (standard IHC practice; UniProt P23771: nuclear localisation). For spatial comparisons, report GATA3-positive nuclei per mm² alongside the total nuclei or eligible cells assessed in each region (standard IHC practice). Normalise to the eligible cell population or sampled tissue area, and apply the same counterstain, threshold and scoring rules across sections (standard IHC practice). Report cytoplasmic colour separately (standard IHC practice).
How can I distinguish genuine GATA3 staining from artefact?
Prioritise well-defined nuclear staining in intact cells because GATA3 is nuclear and HPA reports nuclear expression in selected tissues (UniProt P23771: nucleus; HPA: tissue IHC profile). Check whether staining occurs in plausible cell populations: HPA reports signal in breast glandular cells and kidney glomerular cells, while the selected product example shows human mammary cancer tissue (HPA: tissue IHC; datasheet A00593-1). Question signal limited to section edges, necrotic areas or cytoplasm, and compare it with matched no-primary and detection controls (standard IHC practice; UniProt P23771: nuclear localisation). Investigate endogenous enzyme signal when DAB persists in controls, and avoid treating every positive nucleus as a specific result because presumed off-target binding was reported in the HPA assessment (standard IHC practice; HPA: reliability description).
Boster reagents
Best GATA3 / Trans-acting T-cell-specific transcription factor GATA-3 IHC Antibodies
Catalog IHC images cover human tumors, rat intestine and brain, and mouse brain; IF images cover HeLa cells and rat and mouse brain (catalog IHC/IF captions).
Real IHC data
Anti-GATA3 Antibody ®
Cat # A00593-1
Real IHC data
Anti-GATA3 Rabbit Monoclonal Antibody
Cat # M00593
Real IHC data
Anti-GATA3 Antibody
Cat # A00593
Real IHC data
Anti-Phospho-GATA3 (S308) Rabbit Monoclonal Antibody
Cat # P00593
A00593-1 shows human mammary cancer paraffin-section IHC and HeLa-cell IF (A00593-1 IHC/IF captions); A00593 shows rat and mouse brain IHC and IF (A00593 IHC/IF captions). M00593 IHC captions depict rat intestine and human pituitary tumor, thyroid cancer, and small cell lung cancer, although its listed reactivity is Human only (M00593 IHC captions and catalog reactivity); P00593 shows paraffin-section IHC in human bladder cancer (P00593 IHC caption).
Which to pick: For human tissue IHC, choose A00593-1: its paraffin-section caption documents EDTA retrieval at pH 8.0 and a 1 μg/mL primary concentration; the fixative is unreported (A00593-1 IHC caption). For human IF/ICC, A00593-1 lists both applications and has a HeLa IF image (A00593-1 applications and IF caption); M00593 is a rabbit monoclonal with ICC/IF listed but no IF image caption (M00593 catalog). For mouse or rat tissue IHC and IF, choose A00593, which lists both species and shows brain staining in both assays (A00593 reactivity and IHC/IF captions); P00593 is a phospho-GATA3 (S308) IHC option with a human bladder cancer image (P00593 title, applications and IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P23771 (GATA3_HUMAN, Trans-acting T-cell-specific transcription factor GATA-3).
- Human Protein Atlas. GATA3 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. GATA3 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. GATA3 antibody validation summary (2 antibodies).
- Gene regulatory mechanisms guiding bifurcation of inhibitory and excitatory neuron lineages in the mouse anterior brainstem. eLife 2025 — PMC12431775.
- Expression Patterns of GATA3 in Classical Hodgkin Lymphoma: A Clinico-Pathological Study. Diseases (Basel, Switzerland) 2024 — PMC10969368.
- Diagnostic and Prognostic Roles of GATA3 Immunohistochemistry in Urothelial Carcinoma. Medicina (Kaunas, Lithuania) 2023 — PMC10456966.
- Prevalence and Pattern of GATA3 Immunohistochemical Expression in Female Genital Tract Adenocarcinomas. Iranian journal of pathology 2024 — PMC11304466.
- PubMed PMID:2050118 — UniProt-cited evidence.
- PubMed PMID:1827068 — UniProt-cited evidence.
- PubMed PMID:2017177 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GATA3 (Trans-acting T-cell-specific transcription factor GATA-3). Expected localisation Nuclear staining in selected tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A00593-1). Validated IHC antibody: A00593-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in glomeruli); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human mammary cancer tissue; fixative not specified (datasheet A00593-1); 2. Fixation: Image fixative and duration unreported (datasheet A00593-1); 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 A00593-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A00593-1); 8. Primary antibody: Rabbit anti-GATA3, 0.5-1μg/ml (datasheet A00593-1); 9. Primary incubation: Overnight at 4 °C (datasheet A00593-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A00593-1); Caveat: Presumed off-target staining can complicate interpretation (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unknown (UniProt); Regulation: IL33-linked macrophage response (UniProt). Evidence: UniProt P23771, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm., Datasheet A00593-1, PMC10969368. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GATA3/immunohistochemistry-ihc-guide.