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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 in paraffin sections around the expected nuclear pattern (HPA tissue IHC). Compare kidney glomeruli, where staining is high, with adipocytes, where it is not detected, and assess unexpected signal in light of reported 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 | Nuclear signal in parathyroid glandular and skin basal cells (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 | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target staining was observed (HPA tissue IHC) | |
| Regulation | Acts downstream of IL33 after injury (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended GATA3 IHC & IF Protocols
The catalog antibody’s EDTA retrieval protocol (datasheet A00593-1) is accompanied by published GATA3 IHC methods for lymphoma and genital tract tumors (PMC10969368; PMC11304466).
| 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 noteTry heat-mediated EDTA pH 8.0 retrieval first (datasheet A00593-1). The genital tract study also used EDTA but did not report its pH (PMC11304466).
Section 2
What Is the Expected GATA3 Staining Pattern?
GATA3 is a nuclear transcription factor with no transmembrane segment (UniProt P23771: subcellular location and topology). In paraffin-section IHC, expect staining in selected cell populations, including kidney glomerular cells, breast glandular cells and skin basal-layer cells (HPA: tissue IHC). HPA rates the tissue profile “Enhanced,” while reporting medium consistency with RNA data and presumed off-target binding that was disregarded (HPA: tissue IHC reliability).
| Distinct nuclear staining in the expected cells; kidney glomerular cells are strongly stained, while breast glandular cells show moderate staining (HPA: tissue IHC). | This supports the expected IHC pattern when the nuclear signal is confined to the relevant cells (UniProt P23771: nucleus; HPA: tissue IHC). Compare each cell population with its own HPA reference level; a moderate breast signal need not match the strong kidney signal (HPA: tissue IHC). |
| Predominantly cytoplasmic, membranous or extracellular color, with little convincing nuclear signal. | This is discordant with GATA3’s nuclear location and lack of a transmembrane segment (UniProt P23771: location and topology). Treat it as a possible staining artefact and inspect controls before interpreting it as target expression (general IHC practice). |
| Prominent staining in an unexpected cell population, such as adipocytes or adrenal glandular cells (HPA: both not detected in tissue IHC). | Consider cross-reactivity or endogenous detection activity (general IHC practice). HPA also reports presumed off-target binding in its tissue assessment; its “not detected” entries describe observed staining, not an absolute guarantee of absence (HPA: tissue IHC). |
| Weak color spread across nuclei, cytoplasm and surrounding tissue, without clear cell-specific nuclear contrast. | Diffuse background prevents a confident localization call (general IHC practice). The expected target location is nuclear, and HPA describes expression in selected tissues rather than uniform tissue-wide staining (UniProt P23771: nucleus; HPA: tissue IHC profile). |
| No nuclear staining in a kidney section’s glomerular cells (HPA: high tissue-IHC staining). | A missing signal in this known-positive population calls the assay result into question (HPA: tissue IHC; general IHC practice). Review slide quality, controls and detection conditions before calling another sample negative; HPA’s medium RNA–staining consistency also limits how broadly its tissue observations should be extrapolated (HPA: reliability). |
💡Expected GATA3 appearanceCall a result positive when the expected cells show clear nuclear staining—high in kidney glomerular cells or medium in breast glandular and skin basal-layer cells (HPA: tissue IHC)—while prominent cytoplasmic or unrelated-cell staining raises concern for artefact (UniProt P23771: nucleus; general IHC practice).
| Cell population and tissue | Expected intensity depends on the sampled cells: HPA reports high staining in kidney glomerular cells; medium in breast glandular cells, parathyroid glandular cells and skin basal-layer cells; and no detection in adipocytes (HPA: tissue IHC). Score the relevant cells, rather than averaging staining across the whole section (general IHC practice). |
| Antibody validation | The tissue-IHC profile has an Enhanced reliability label, with medium consistency against RNA data and presumed off-target binding disregarded (HPA: tissue IHC reliability). CAB016217 is IHC Enhanced, whereas HPA029730 has ICC Supported status with no IHC status listed; these are assay-specific validation entries (HPA: antibodies). |
| Nuclear location and topology | GATA3 is nuclear and has no transmembrane segment (UniProt P23771: location and topology). Nuclear contrast therefore carries the interpretation; a membrane outline alone does not match the supplied target-location evidence (UniProt P23771: location and topology). |
| Isoforms and processing | UniProt lists 2 isoforms, a single chain spanning residues 1–443, and no signal peptide or propeptide (UniProt P23771: isoforms and processing). No epitope map is supplied, so these annotations cannot establish whether a particular antibody detects both isoforms or predict a section-specific staining difference. |
| Endogenous detection activity | Endogenous enzyme activity can create chromogenic signal independently of primary-antibody binding (general IHC practice). Check the detection system and an appropriate negative control if unexpected color appears; this is a general assay concern, not a GATA3-specific effect established by HPA or UniProt. |
| IF/ICC expectation? | For a separate IF/ICC assay, the expected location is nucleoplasmic; HPA marks it supported and lists images from A-431, U-251MG and U2OS cells (HPA: subcellular ICC-IF). This localization answers the IF/ICC pattern question; the IHC tissue intensity entries do not specify an IF/ICC protocol (HPA: tissue IHC and subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| Kidney glomerular cells lack visible nuclear stain (HPA: high tissue-IHC staining). | The staining run may have inadequate signal or an unsuitable control outcome (general IHC practice); the supplied sources do not identify a GATA3-specific fixation sensitivity. | Check section integrity, positive-control performance, primary-antibody application and chromogenic detection steps (general IHC practice). Repeat with a validated positive section before scoring the test section negative (HPA: kidney tissue IHC; general IHC practice). |
| Color is mainly cytoplasmic or membranous. | The compartment conflicts with the supplied nuclear localization and absent transmembrane segment (UniProt P23771: location and topology); nonspecific detection is possible (general IHC practice). | Inspect the nuclear counterstain and an appropriate negative control, then review blocking, washing and detection conditions (general IHC practice). Record only clearly localized nuclear signal as supporting the expected pattern (UniProt P23771: nucleus). |
| Adipocytes or adrenal glandular cells show conspicuous staining (HPA: not detected in tissue IHC). | Cross-reactivity or endogenous detection activity is plausible (general IHC practice). HPA’s reliability note reports presumed off-target binding that was disregarded (HPA: tissue IHC reliability). | Compare the suspect cells with a positive nuclear control and a detection negative control; assess whether color tracks cell type or processing background (general IHC practice). Treat the unexpected result cautiously against the HPA tissue observations. |
| The entire section has low-contrast brown haze. | Diffuse chromogen or nonspecific background can obscure nuclear localization (general IHC practice). | Review blocking, washes, primary-antibody concentration and chromogen development using the assay’s validated instructions (general IHC practice). Reassess only after nuclei can be distinguished from the surrounding tissue (UniProt P23771: nucleus; general IHC practice). |
| Breast or skin looks weaker than kidney. | The supplied reference levels differ: breast glandular and skin basal-layer cells are medium, while kidney glomerular cells are high (HPA: tissue IHC). | Score the appropriate cell population against its tissue-specific expectation (HPA: tissue IHC). Check positive controls if the signal is absent, but do not require all three tissues to reach the kidney reference intensity (HPA: tissue IHC). |
| An IF/ICC image appears diffuse despite an IHC-positive control. | HPA supports nucleoplasmic ICC-IF localization, while its tissue intensity observations come from IHC; performance in one assay does not itself establish the other assay’s staining quality (HPA: subcellular ICC-IF and tissue IHC). | For IF/ICC interpretation, look for nucleoplasmic contrast and evaluate that assay’s controls separately (HPA: subcellular ICC-IF; general immunofluorescence practice). Use the separate IF/ICC guide for method choices. |
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: High in cells in glomeruli; UniProt P23771: nucleus). Use adipose tissue as the negative tissue, assessing adipocytes (HPA: Not detected in adipocytes); on the kidney slide, cells without nuclear signal provide an internal background reference, but the supplied HPA row does not establish every nonglomerular cell as negative.
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) and host-matched nonimmune rabbit IgG isotype controls, plus GATA3-knockout material or a peptide-block control if the immunizing peptide is available (selected-SKU caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase and block endogenous biotin in kidney when using the caption’s streptavidin–biotin/DAB detection system (selected-SKU caption: SABC with DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption does not state a fixative (selected-SKU caption: fixative unreported). That caption uses heat retrieval in EDTA at pH 8.0, but does not establish whether retrieval is essential; kidney endogenous biotin may complicate its SABC detection (selected-SKU caption: EDTA retrieval and SABC; standard IHC practice). Whether frozen sections or IF/ICC are easier is unreported; the available ICC-IF evidence supports nucleoplasmic localisation (HPA subcellular: nucleoplasm supported).
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
Troubleshoot GATA3 staining in paraffin sections by checking nuclear localisation, tissue context and method controls before scoring chromogenic signal.
What retrieval should I try when nuclear GATA3 staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0, the condition used for the catalog antibody in paraffin-embedded human mammary cancer tissue (datasheet A00593-1). Keep section thickness, heating and cooling consistent across comparison slides, then check whether nuclei become clearer without increasing diffuse staining (standard IHC practice). The reported workflow used 1 μg/ml primary antibody overnight at 4°C after retrieval, providing a reference point when diagnosing weak signal (datasheet A00593-1). If staining remains weak, compare a separately optimised retrieval condition against EDTA pH 8.0 on matched sections, judging nuclear signal and background together (standard IHC practice).
Could fixation explain variable GATA3 staining between paraffin blocks?
The catalog image reports paraffin-embedded human mammary cancer tissue but does not state its fixative or fixation duration, so GATA3-specific fixation sensitivity is unknown (datasheet A00593-1). Record the fixative, time before fixation and fixation duration for each block, and compare sections processed and stained together (standard IHC practice). If signal varies, hold retrieval at EDTA pH 8.0 and primary incubation at 1 μg/ml overnight at 4°C while assessing the blocks (datasheet A00593-1; standard IHC practice). Include a consistently staining reference section in each run so processing variation can be distinguished from tissue variation (standard IHC practice).
How should I assess apparent cytoplasmic GATA3 staining?
Score convincing GATA3 staining in nuclei: UniProt places the protein in the nucleus, and HPA supports nucleoplasmic localisation (UniProt P23771 subcellular; HPA subcellular). GATA3 has no annotated transmembrane segment, so a dominant membrane rim is inconsistent with its recorded topology (UniProt P23771 topology). Examine cytoplasmic colour at higher magnification beside a haematoxylin counterstain, checking whether pigment or overlapping cells create a false impression of localisation (standard IHC practice). Compare the pattern with an expected nuclear reference, such as glandular cells in breast tissue, reported at medium staining intensity (HPA tissue IHC).
Can isoforms or phosphorylation explain discordant antibody staining?
UniProt lists 2 GATA3 isoforms and phosphorylated residues including Ser115 and Ser162 (UniProt P23771 isoforms; UniProt P23771 modified residues). The supplied catalog caption does not identify the antibody epitope, so it cannot establish isoform recognition or sensitivity to either modification (datasheet A00593-1). When two antibodies disagree, compare their documented immunogens or epitopes and stain adjacent sections under matched retrieval and detection conditions (standard IHC practice). Report the discordance as antibody-dependent until epitope information or an independent specificity control supports a molecular explanation (standard IHC practice).
How can I adapt the GATA3 localisation check to multiplex IF?
For the separate IF assay, evaluate GATA3 in nuclei because UniProt assigns nuclear localisation and HPA supports nucleoplasmic localisation (UniProt P23771 subcellular; HPA subcellular). Multiplex with a cell-identity marker such as CD3 for T cells, and assess GATA3 within marker-positive nuclei rather than across the whole field (UniProt P23771 tissue specificity; standard IF practice). Choose a fluorophore channel after inspecting unstained tissue autofluorescence, with single-stain controls to assess bleed-through (standard IF practice). Because GATA3 lacks a transmembrane segment and is nuclear, optimise gentle permeabilisation for intracellular access, then compare nuclear signal and background with a no-primary control (UniProt P23771 topology; standard IF practice).
What should I check if brown staining obscures GATA3-positive nuclei?
First compare a no-primary control with the test section, and check endogenous peroxidase blocking and DAB development because these affect chromogenic background (standard IHC practice). The catalog workflow used 10% goat serum blocking, a biotinylated secondary for 30 minutes at 37°C, and streptavidin-biotin detection with DAB (datasheet A00593-1). If background persists, inspect whether detection reagents stain tissue without primary antibody and shorten development or adjust primary concentration using matched sections (standard IHC practice). HPA reports presumed off-target binding despite enhanced reliability, so diffuse staining should be judged against nuclear localisation and controls (HPA tissue IHC; UniProt P23771 subcellular).
How should I quantify heterogeneous nuclear GATA3 staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue compartment and eligible cells before scoring, then record the percentage of nuclei at each staining intensity (standard IHC practice). An H-score can sum percentages scored 0–3, yielding 0–300; record both intensity and percentage so the result remains interpretable (standard IHC practice). For sparse populations, report positive nuclei per mm² alongside total eligible nuclei or tissue area, and apply one threshold across slides (standard IHC practice). Keep cell types separate when relevant: HPA reports high staining in kidney glomerular cells and medium staining in breast glandular cells (HPA tissue IHC).
How do I distinguish true GATA3 staining from artefact?
Prioritise crisp nuclear signal in an appropriate cell population because GATA3 is nuclear and HPA describes nuclear expression in selected tissues (UniProt P23771 subcellular; HPA tissue IHC). Check suspicious cytoplasmic or membrane colour, section-edge staining and necrotic areas against adjacent intact tissue before calling cells positive (UniProt P23771 topology; standard IHC practice). Use a no-primary control to identify staining from endogenous enzyme activity or detection reagents, particularly when DAB colour appears outside nuclei (standard IHC practice). Interpret negative cells within tissue context: HPA reports medium staining in breast glandular cells but no detected staining in adipocytes (HPA tissue IHC).
Boster reagents
Best GATA3 / Trans-acting T-cell-specific transcription factor GATA-3 IHC Antibodies
The catalog shows GATA3 IHC images from human, mouse, and rat tissues and IF images from human cells and mouse and rat brain (catalog image 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 IHC in paraffin-embedded human mammary cancer tissue and IF in human cells (A00593-1 image captions); M00593 lists human reactivity and IHC/IF applications, although its displayed IHC image depicts rat intestine (M00593 reactivity, applications, and IHC caption). A00593 shows IHC and IF in mouse and rat brain (A00593 image captions); P00593 shows phospho-GATA3 (S308) IHC in paraffin-embedded human bladder cancer (P00593 IHC caption).
Which to pick: For human paraffin-section IHC, choose A00593-1: its image documents EDTA pH 8 retrieval and 1 μg/mL primary antibody, while the fixative is unreported (A00593-1 IHC caption). For human IF/ICC, A00593-1 has a human-cell IF image and an ICC application listing (A00593-1 IF caption and applications); for mouse or rat tissue IHC and IF, A00593 has images in both species and both applications listed (A00593 image captions and applications). Choose P00593 when staining phospho-GATA3 (S308) in human paraffin sections (P00593 title, reactivity, and IHC caption); treat M00593’s rat-intestine image cautiously because its catalog reactivity lists only human (M00593 IHC caption and reactivity).
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: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Presumed off-target staining was observed (HPA tissue IHC); Isoform / epitope: 2 isoforms ; epitope coverage is unknown (UniProt); Regulation: Acts downstream of IL33 after injury (UniProt). Evidence: UniProt P23771, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm., Datasheet A00593-1, PMC10969368, PMC11304466. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GATA3/immunohistochemistry-ihc-guide.