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GADD45B / Growth arrest and DNA damage-inducible protein GADD45 beta · IHC design guide
Design Immunohistochemistry for GADD45B
Plan GADD45B chromogenic IHC in paraffin sections using the catalog antibody's validated conditions. Compare cytoplasmic staining with placental decidual cells, where staining is high, while checking specificity because staining and RNA data have low concordance (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GADD45B 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 | General cytoplasmic staining reported (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining; high in placental decidual cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03508-1) | |
| Positive control | Placenta+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 A03508-1) | |
| Caveat | Staining has low concordance with RNA; verify specificity (HPA tissue IHC) | |
| Regulation | Stress-response role; induction unreported (UniProt) | |
| Isoform / epitope | One 1–160 chain; no isoforms annotated (UniProt) |
Section 1
Recommended GADD45B IHC & IF Protocols
Compare the catalog antibody protocol (datasheet: A03508-1) with published IHC procedures for retina, lung, colorectal tissue microarrays, and kidney (PMC2662500; PMC11785585; PMC6071283; PMC7508261).
| Sample | Paraffin-embedded human placenta tissue; fixative not specified (datasheet A03508-1) |
| Fixation | Image fixative and duration unreported (datasheet A03508-1); 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 A03508-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03508-1) |
| Primary antibody | Rabbit anti-GADD45B, 2 μg/ml (datasheet A03508-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03508-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03508-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GADD45B-positive staining in decidual cells of placenta (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet: A03508-1); the cited kidney study reports antigen repair without specifying its conditions (PMC7508261).
Section 2
What Is the Expected GADD45B Staining Pattern?
For paraffin-section IHC, expect GADD45B staining mainly in the cytoplasm, with particularly strong staining in placental decidual cells and medium staining in several other listed cell types (HPA tissue IHC). GADD45B has no transmembrane segment (UniProt O75293 topology). HPA rates its tissue IHC as Approved, but reports low consistency with RNA expression and pending external verification (HPA tissue IHC).
| Distinct cytoplasmic staining in placental decidual cells, stronger than nearby background. | This matches the reported high decidual-cell signal and general cytoplasmic pattern (HPA tissue IHC). Score the identified cells and their staining intensity; a uniformly colored section gives less support for cell-specific staining (general IHC practice). HPA's Approved rating remains qualified by low agreement with RNA data and pending external verification (HPA tissue IHC). |
| Predominantly nuclear staining in paraffin-section IHC, with little discernible cytoplasmic signal. | Recheck the result against HPA's general cytoplasmic tissue-IHC profile (HPA tissue IHC). Nuclear staining alone is not proof of artefact: HPA's separate ICC-IF observations place GADD45B mainly in the nucleoplasm (HPA subcellular ICC-IF). Resolve the discrepancy with appropriate controls before calling the tissue pattern positive (general IHC practice). |
| A sharp membrane outline or strong staining in cells outside the reported positive populations. | A membrane-only outline does not fit the tissue-IHC profile (HPA tissue IHC) or the absence of a transmembrane segment (UniProt O75293 topology). Unexpected cell staining may reflect nonspecific binding or endogenous chromogenic activity (general IHC practice); cell identity and control slides should guide interpretation. |
| Diffuse brown color across tissue compartments, including areas without distinct cells. | Treat widespread background as uninterpretable until checked with a no-primary control and inspection for endogenous enzyme activity or pigment (general IHC practice). HPA describes a cellular, generally cytoplasmic pattern (HPA tissue IHC); diffuse color by itself does not establish GADD45B localization. |
| No discernible staining in placental decidual cells on an otherwise interpretable section. | This conflicts with HPA's reported high decidual-cell staining (HPA tissue IHC), but a single negative run does not establish absence of GADD45B. Check tissue identity and the IHC detection workflow with controls (general IHC practice). HPA's tissue observations have low RNA agreement and await external verification (HPA tissue IHC). |
💡Expected GADD45B appearanceA convincing positive is cell-resolved, predominantly cytoplasmic staining that is strong in placental decidual cells (HPA tissue IHC); a membrane-only rim or diffuse, cell-independent brown deposit is suspicious (UniProt O75293 topology; general IHC practice).
| Tissue and cell selection | Placental decidual cells have high reported staining; adipocytes, selected glandular cells, hematopoietic cells, respiratory epithelial cells and caudate glia have medium staining (HPA tissue IHC). Identify the scored population before comparing intensity (general IHC practice). |
| Lower-staining comparison tissue | Hippocampal glia, parathyroid glandular cells, ovarian follicle cells, smooth muscle cells and splenic red-pulp cells are listed as low (HPA tissue IHC). Low is not a documented absence, and HPA supplies no negative tissue in this payload (HPA tissue IHC). |
| Topology and tissue-IHC localization | The single annotated chain spans residues 1–160; no signal peptide, propeptide, transmembrane segment or glycosylation site is listed (UniProt O75293). These annotations support scrutiny of a membrane-only pattern but do not establish fixation sensitivity or an antigen-retrieval condition (UniProt O75293 topology; general IHC practice). |
| Strength of tissue evidence | HPA marks tissue IHC Approved while reporting low consistency between antibody staining and RNA expression and pending external verification (HPA tissue IHC). Its reported intensity is a reference pattern, not a guaranteed result in every specimen (HPA tissue IHC; general IHC practice). |
| IF/ICC Q: Should nuclear IF signal be scored as wrong? | A: No. HPA reports mainly nucleoplasmic ICC-IF localization, with additional nucleolar and cytosolic signal (HPA subcellular ICC-IF). Interpret those images within IF/ICC; HPA's paraffin-tissue IHC profile is generally cytoplasmic (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Placental decidual cells appear negative. | The positive population may be misidentified, or the IHC run may have failed (HPA tissue IHC; general IHC practice). | Confirm decidual-cell identity and examine a run control and no-primary control; review the catalog antibody's IHC-P directions for the actual assay settings (general IHC practice). Do not infer a GADD45B-specific fixation effect from this result. |
| Signal is weak in a tissue expected to show medium staining. | HPA's medium category is cell-specific, and its tissue-IHC observations have low RNA agreement (HPA tissue IHC). | Locate the listed cell population, compare it with a placental decidual-cell positive control, and score signal relative to local background (HPA tissue IHC; general IHC practice). |
| Only nuclei stain in paraffin-section IHC. | The result differs from HPA's general cytoplasmic tissue pattern, although nuclear ICC-IF signal is reported (HPA tissue IHC; HPA subcellular ICC-IF). | Check staining controls and cell identity, then report the assay-specific discrepancy. Do not transfer the ICC-IF localization directly into a tissue-IHC scoring rule (general IHC practice). |
| Cells show a crisp membrane-only rim. | This conflicts with the reported cytoplasmic tissue pattern and lack of a transmembrane segment (HPA tissue IHC; UniProt O75293 topology). | Inspect the no-primary control and neighboring tissue for edge deposits or nonspecific staining; withhold a positive call unless a cellular pattern is supported (general IHC practice). |
| Unexpected cell types stain or brown signal spreads across the section. | Nonspecific binding, pigment or endogenous chromogenic activity can mimic cellular signal (general IHC practice). | Compare no-primary and detection controls, assess staining against the counterstain, and score only identifiable cells with localized signal (general IHC practice). HPA does not list a negative tissue here (HPA tissue IHC). |
| Low-listed cells stain faintly. | HPA labels these populations low, not negative; the tissue-IHC evidence also awaits external verification (HPA tissue IHC). | Record cell identity, intensity and background rather than treating faint signal as automatically false or confirmed positive (general IHC practice). Compare with the high decidual-cell reference pattern (HPA tissue IHC). |
Experiment evidence
IHC & IF images showing GADD45B staining
A03508-1 · IHCIHC analysis of GADD45B using anti-GADD45B antibody (A03508-1). GADD45B was detected in a paraffin-embedded section of human placenta tissue
A03508-1 · IHCIHC analysis of GADD45B using anti-GADD45B antibody (A03508-1). GADD45B was detected in a paraffin-embedded section of human spleen tissue.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000099860-GADD45B/tissue/cerebral+cortex#imid_7436936
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000099860-GADD45B/tissue/colon#imid_7426062
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000099860-GADD45B/tissue/liver#imid_7426063
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000099860-GADD45B/tissue/kidney#imid_7426056
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029816; green = GADD45B, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029816; green = GADD45B, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GADD45B IHC & IF
🧪Run placenta first and assess decidual cells for GADD45B staining (HPA: High in decidual cells). HPA detects GADD45B in all 45 scored tissues, so there is no validated negative tissue; use no-primary and isotype controls, and treat cells showing only counterstain on the positive slide as background references rather than proven target-negative cells (HPA: no negative tissue rows; standard IHC practice).
Positive control tissue: Placenta (Decidual cells, HPA High)
Negative control tissue: None in HPA: GADD45B 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 GADD45B in A-431, U2OS, U-251MG, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide; a concentration-matched rabbit IgG isotype control matched to the primary antibody’s clonality; and, if available, a GADD45B knockout specimen or peptide-blocked primary as a biological specificity control (selected-SKU caption: rabbit primary; standard IHC practice). For placenta, quench endogenous peroxidase before chromogenic detection and check tissue autofluorescence if using IF (standard IHC/IF 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). The caption reports heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required under other conditions (selected-SKU caption). The supplied evidence does not establish whether frozen sections or IF/ICC are easier than paraffin IHC; placenta background from endogenous peroxidase or autofluorescence should be checked with the controls above (standard IHC/IF practice).
HPA tissue IHC evidence for GADD45B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Placenta | Decidual cells | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: GADD45B is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced GADD45B IHC Tips
Troubleshoot GADD45B staining in paraffin sections by checking retrieval, controls, cell identity and compartment before interpreting DAB signal.
What should I change when GADD45B staining is weak after antigen retrieval?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin-section IHC (datasheet A03508-1). If staining is weak, compare retrieval heating and cooling conditions across adjacent sections while keeping antibody concentration and DAB development constant (standard IHC practice). The catalog image used 2 μg/ml primary antibody overnight at 4°C, so it provides a useful starting point for that comparison (datasheet A03508-1). Check tissue integrity and a no-primary control before attributing a faint signal to inaccessible GADD45B epitopes (standard IHC practice). Try another retrieval buffer only as a documented fallback if the EDTA condition remains weak (standard IHC practice).
Could fixation explain variable GADD45B staining between paraffin sections?
Target-specific sensitivity of GADD45B to fixation is unknown from the supplied evidence; the catalog paraffin-section caption does not report its fixative (datasheet A03508-1). Record fixative identity, fixation duration and processing history for each specimen before comparing staining intensity (standard IHC practice). For troubleshooting, stain sections from differently processed blocks together using the same pH 8.0 EDTA retrieval and detection conditions (datasheet A03508-1; standard IHC practice). Inspect morphology alongside signal, because poor preservation can complicate identification of stained cells (standard IHC practice). Do not infer a fixation effect from tissue staining patterns or from the absence of annotated GADD45B modifications (HPA tissue IHC; UniProt O75293).
How should I assess cytoplasmic versus nuclear GADD45B staining?
Evaluate chromogenic staining by compartment within identifiable cells: the tissue IHC profile describes general cytoplasmic expression (HPA tissue IHC). Nuclear signal warrants separate review because cell imaging places GADD45B mainly in the nucleoplasm, with additional nucleolar and cytosolic localisation (HPA subcellular). The protein has no annotated transmembrane segment, so a sharp membrane-only pattern needs independent validation (UniProt O75293 topology; standard IHC practice). Compare stained cells with the counterstain and tissue morphology, then record nuclear and cytoplasmic scores separately rather than pooling them (standard IHC practice). Treat a compartment mismatch as a prompt to check controls and antibody specificity, not as proof of absence (standard IHC practice).
Could an isoform or masked epitope account for discordant staining?
No GADD45B isoforms, domains or modified residues are annotated in the supplied record, so those entries cannot explain a particular staining pattern (UniProt O75293). The record describes a single 1–160 chain and no signal peptide or transmembrane segment (UniProt O75293). Because the antibody epitope is unspecified here, do not assign an unexpected nuclear or cytoplasmic pattern to a known epitope position (datasheet A03508-1; standard IHC practice). Compare retrieval conditions on adjacent sections and include a no-primary control when testing whether masking or detection contributes to the discrepancy (standard IHC practice). An independently validated antibody to a distinct epitope can add specificity evidence if available (standard IHC practice).
How can IF help resolve ambiguous GADD45B localisation in the same tissue?
Use IF as a complementary localisation check: cell imaging reports mainly nucleoplasmic GADD45B, with additional nucleolar and cytosolic signal (HPA subcellular). Multiplex with a validated marker for the cell type under examination; decidual cells are a relevant example in placenta (HPA tissue IHC; standard IF practice). Check unstained tissue autofluorescence and choose spectrally separated fluorophores with adequate contrast before assigning puncta or diffuse signal to GADD45B (standard IF practice). Because GADD45B has no annotated transmembrane segment and nuclear access may matter, assess permeabilisation for intracellular epitope access (UniProt O75293 topology; standard IF practice). Include single-colour and no-primary controls to distinguish bleed-through and background (standard IF practice).
What should I check when DAB staining is diffuse or widespread?
Run a no-primary section to identify signal from the detection system, and inspect tissue edges and damaged areas separately (standard IHC practice). The catalog image used a peroxidase-conjugated secondary and DAB, making endogenous peroxidase blocking a relevant general workflow check (datasheet A03508-1; standard IHC practice). Its section was blocked with 10% goat serum, and the primary was applied at 2 μg/ml overnight at 4°C (datasheet A03508-1). If background persists, compare primary-antibody titrations while holding retrieval, blocking and DAB development constant (standard IHC practice). Reassess cellular detail before calling uniform colour a GADD45B-positive pattern (standard IHC practice).
How should I quantify GADD45B staining across heterogeneous paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; HPA reports high staining in placental decidual cells and a general cytoplasmic tissue pattern (HPA tissue IHC). For a defined cell population, report the percentage positive and staining intensity, or calculate an H-score on a 0–300 scale (standard IHC practice). Normalise positive-cell counts to the number of eligible cells, or area-based counts to evaluable tissue area in mm² (standard IHC practice). Exclude folds, necrosis and section edges by a rule set before review, and apply the same threshold across specimens (standard IHC practice). Report nuclear and cytoplasmic scores separately when both are assessed (standard IHC practice).
When is apparent GADD45B positivity more likely to be artefact?
A credible call should follow cell boundaries and tissue morphology; high staining in placental decidual cells provides one comparison, although staining and RNA show low consistency (HPA tissue IHC; standard IHC practice). Review isolated membrane-only staining critically because GADD45B has no annotated transmembrane segment (UniProt O75293 topology; standard IHC practice). Compare compartment patterns carefully: tissue IHC is generally cytoplasmic, while cell imaging places GADD45B mainly in the nucleoplasm (HPA tissue IHC; HPA subcellular). Edge-restricted colour, necrotic areas or signal in the no-primary control support an artefact check rather than a positive call (standard IHC practice). Check endogenous peroxidase blocking when unexplained DAB signal persists (standard IHC practice).
Boster reagents
Best GADD45B / Growth arrest and DNA damage-inducible protein GADD45 beta IHC Antibodies
The catalog antibody has real IHC data from human paraffin sections of placenta, spleen, renal cell carcinoma, and tonsil (catalog image captions); IF/ICC validation is unlisted (catalog applications).
Real IHC data
Anti-GADD45B Antibody ®
Cat # A03508-1
A03508-1 is the only SKU and is listed for human IHC (catalog applications and reactivity). Its IHC captions show staining in paraffin sections of human placenta, spleen, renal cell carcinoma, and tonsil (catalog image captions).
Which to pick: Choose A03508-1 for human paraffin-section IHC; it is a rabbit polyclonal antibody with IHC listed and a paraffin-section figure (catalog host, dilution information, applications, and image caption). The caption does not report the fixative (catalog image caption). No listed SKU has IF/ICC validation or reactivity beyond human, so the payload supports no IF/ICC or cross-species pick (catalog applications and reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75293 (GA45B_HUMAN, Growth arrest and DNA damage-inducible protein GADD45 beta).
- Human Protein Atlas. GADD45B tissue IHC expression (reliability: Approved).
- Human Protein Atlas. GADD45B subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nucleoli and cytosol..
- Human Protein Atlas. GADD45B antibody validation summary (2 antibodies).
- Growth arrest and DNA damage protein 45b (Gadd45b) protects retinal ganglion cells from injuries. Neurobiology of disease 2009 — PMC2662500.
- DNA hypomethylation-mediated upregulation of GADD45B facilitates airway inflammation and epithelial cell senescence in COPD. Journal of advanced research 2025 — PMC11785585.
- GADD45B as a Prognostic and Predictive Biomarker in Stage II Colorectal Cancer. Genes 2018 — PMC6071283.
- GADD45B Promotes Glucose-Induced Renal Tubular Epithelial-Mesenchymal Transition and Apoptosis via the p38 MAPK and JNK Signaling Pathways. Frontiers in physiology 2020 — PMC7508261.
- PubMed PMID:9827804 — UniProt-cited evidence.
- PubMed PMID:10773677 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GADD45B (Growth arrest and DNA damage-inducible protein GADD45 beta). Expected localisation General cytoplasmic staining reported (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03508-1). Validated IHC antibody: A03508-1 at 2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Placenta (Decidual cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human placenta tissue; fixative not specified (datasheet A03508-1); 2. Fixation: Image fixative and duration unreported (datasheet A03508-1); 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 A03508-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03508-1); 8. Primary antibody: Rabbit anti-GADD45B, 2 μg/ml (datasheet A03508-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03508-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03508-1); Caveat: Staining has low concordance with RNA; verify specificity (HPA tissue IHC); Isoform / epitope: One 1–160 chain; no isoforms annotated (UniProt); Regulation: Stress-response role; induction unreported (UniProt). Evidence: UniProt O75293, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A03508-1, PMC2662500, PMC11785585. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GADD45B/immunohistochemistry-ihc-guide.