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RXRA / Retinoic acid receptor RXR-alpha · IHC design guide
Design Immunohistochemistry for RXRA
Plan RXRA chromogenic IHC in paraffin sections around its ubiquitous nuclear staining pattern (HPA tissue IHC). This guide covers the catalog antibody’s 0.5–1 μg/mL IHC range (datasheet A01299-1) and consistent fixation (standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RXRA Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Ubiquitous nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | Widespread nuclear staining across tissue cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A01299-1) | |
| Positive control | Adrenal gland+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 A01299-1) | |
| Caveat | Widespread nuclear staining limits tissue specificity (HPA tissue IHC) | |
| Regulation | Staining-intensity regulation is unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences are unspecified (UniProt) |
Section 1
Recommended RXRA IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A01299-1) is accompanied by a published human nucleus pulposus IHC protocol (PMC12995935 methods).
| Sample | Paraffin-embedded mouse liver tissue; fixative not specified (datasheet A01299-1) |
| Fixation | Image fixative and duration unreported (datasheet A01299-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A01299-1) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01299-1) |
| Primary antibody | Rabbit anti-RXRA, 0.5-1μg/ml (datasheet A01299-1) |
| Primary incubation | Overnight at 4 °C (datasheet A01299-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01299-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RXRA-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet: A01299-1); the published protocol also used citrate buffer but did not report its pH (PMC12995935 methods).
Section 2
What Is the Expected RXRA Staining Pattern?
RXRA is expected mainly in nuclei across many tissues (HPA: ubiquitous nuclear expression; HPA: IHC Supported). Look for nuclear staining in glandular, epithelial, hematopoietic, adipocyte, glial, and cerebellar granular-layer cells represented in the HPA tissue survey (HPA: High in listed cell types). UniProt also lists cytoplasm and mitochondrion, and reports no transmembrane segment (UniProt P19793: subcellular location and topology). HPA notes medium agreement with RNA data and pending external verification (HPA: reliability description).
| Distinct nuclear stain in adrenal glandular cells or bronchial respiratory epithelium. | This fits the expected compartment and two HPA High cell populations (HPA: ubiquitous nuclear expression; HPA: High in adrenal glandular and bronchial respiratory epithelial cells). Compare nuclear signal with adjacent background before scoring (standard IHC practice). |
| Predominant membrane, extracellular, or structureless cytoplasmic stain obscures nuclei. | Question specificity or detection background: RXRA has no transmembrane segment, and the surveyed IHC pattern is nuclear (UniProt P19793: topology; HPA: tissue profile). Cytoplasmic signal alone is not proof of artefact because UniProt also lists cytoplasm (UniProt P19793: subcellular location). |
| Strong stain appears in an unexpected cell population while listed positive cells are weak. | Check cross-reactivity and endogenous detection activity with suitable controls (standard IHC practice). Do not call that population RXRA-negative solely from this survey: HPA reports ubiquitous nuclear expression and supplies no negative tissue list (HPA: tissue profile; HPA: negative list empty). |
| Diffuse stain covers nuclei, cytoplasm, and tissue background without crisp boundaries. | The distribution cannot be scored as convincing nuclear RXRA (HPA: ubiquitous nuclear expression). Nonspecific antibody binding, incomplete blocking, or detection background are possible workflow causes; an omitted-primary control helps separate detection background from primary-dependent stain (standard IHC practice). |
| No nuclear signal in a listed High population, such as bone marrow hematopoietic cells. | Treat the run as potentially insensitive before inferring absence of RXRA (HPA: High in bone marrow hematopoietic cells). Verify that an appropriate positive control stained and that retrieval, primary incubation, and detection ran as intended (standard IHC practice). |
💡Expected RXRA appearanceCall a section positive when discrete nuclear stain is readily visible in an HPA-listed High cell population, such as appendix glandular cells, with low surrounding background; isolated membrane or extracellular stain is an unconvincing positive pattern (HPA: ubiquitous nuclear expression; HPA: High in appendix glandular cells; UniProt P19793: topology; standard IHC practice).
| Cell compartment | HPA tissue IHC reports ubiquitous nuclear expression, while UniProt also lists cytoplasm and mitochondrion (HPA: tissue profile; UniProt P19793: subcellular location). Score the nuclear pattern first; assess additional compartments separately rather than treating every nonnuclear pixel as equivalent evidence. |
| Cell population and confidence | HPA lists High staining in several sampled cell types and low tissue specificity at the RNA level (HPA: positive list; HPA: RNA specificity). Its IHC reliability is Supported, with medium RNA agreement and external verification pending; these entries guide control choice, not a universal intensity threshold (HPA: reliability). |
| Antigen retrieval | For paraffin IHC, compare retrieval conditions against a known-positive section if nuclear signal is weak (standard IHC practice). The supplied HPA and UniProt records give no RXRA-specific retrieval condition or fixation-sensitivity result (HPA: tissue profile; UniProt P19793 record). |
| Isoforms and epitope | UniProt lists two RXRA isoforms and a nuclear-receptor ligand-binding domain at residues 227–458 (UniProt P19793: isoforms and domain). Without an antibody epitope in the supplied evidence, neither isoform coverage nor domain-specific staining can be predicted from these facts alone. |
| Modified residues and processing | UniProt lists phosphorylated residues, one chain spanning residues 1–462, and no signal peptide or propeptide (UniProt P19793: modified residues and processing). These annotations do not establish whether the IHC antibody recognizes a modified site or predict a tissue-specific staining change without epitope evidence. |
| IF/ICC Q&A | Q: What localization is expected on the separate IF/ICC guide? A: Mainly nucleoplasm, with additional Golgi localization reported in A-431 and U2OS image data (HPA: subcellular ICC-IF summary and image cell lines). This IF observation does not set an IHC staining threshold. |
| Situation | Likely cause | Next action |
|---|---|---|
| A listed High tissue shows weak or absent nuclear staining. | The run may lack sensitivity; tissue absence alone is unsupported for that listed population (HPA: High positive list; standard IHC practice). | Inspect a positive control in the same run, then review section processing, retrieval, primary incubation, and detection steps (standard IHC practice). Do not assign an RXRA-specific retrieval recipe from these records. |
| Stain is diffuse across tissue and blank spaces. | Background from antibody binding or the chromogenic detection workflow may overwhelm compartment detail (standard IHC practice). | Review blocking, washing, primary concentration, and development time; compare an omitted-primary control (standard IHC practice). Accept a result only when nuclei can be distinguished from background (HPA: nuclear tissue profile). |
| Unexpected cells stain strongly while expected nuclei remain faint. | Cross-reactivity or endogenous detection activity is possible; the HPA survey does not define a negative cell population (HPA: tissue profile and negative list; standard IHC practice). | Compare morphology and compartment with a listed High control; check omitted-primary and detection controls before assigning cell identity or specificity (standard IHC practice). |
| Only membrane or extracellular outlines are prominent. | That distribution conflicts with HPA nuclear IHC and UniProt's lack of a transmembrane segment (HPA: tissue profile; UniProt P19793: topology). | Reassess background and morphology, compare a positive nuclear control, and avoid scoring outlines as RXRA without independent support (standard IHC practice). |
| Some cytoplasmic staining accompanies clear nuclear staining. | UniProt includes cytoplasm among RXRA locations, but HPA tissue IHC describes a ubiquitous nuclear pattern (UniProt P19793: subcellular location; HPA: tissue profile). | Score the interpretable nuclear component separately; compare cytoplasmic signal with background and controls before describing it as specific (standard IHC practice). |
| Two IHC antibodies yield different nuclear intensity. | Both listed antibodies have Supported IHC validation; the supplied status does not establish identical intensity or epitope recognition (HPA: CAB004565 and CAB005352 IHC status). | Compare compartment, cell population, and controls under each antibody's validated conditions; report the discrepancy rather than inferring a biological change from intensity alone (HPA: antibody status; standard IHC practice). |
Experiment evidence
IHC & IF images showing RXRA staining
A01299-1 · IHCIHC analysis of RXRA using anti-RXRA antibody (A01299-1). RXRA was detected in paraffin-embedded section of mouse liver tissue. Heat mediate
A01299-1 · IHCIHC analysis of RXRA using anti-RXRA antibody (A01299-1). RXRA was detected in paraffin-embedded section of rat liver tissue. Heat mediated
A01299-1 · IFIF analysis of RXRA using anti-RXRA antibody (A01299-1). RXRA was detected in immunocytochemical section of A431 cells. Enzyme antigen retri
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000186350-RXRA/tissue/cerebral+cortex#imid_1545608
HPA · EsophagusHuman Protein Atlas IHC, Esophagus. Source: https://www.proteinatlas.org/ENSG00000186350-RXRA/tissue/esophagus#imid_1545588
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000186350-RXRA/tissue/colon#imid_1545534
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000186350-RXRA/tissue/liver#imid_1545543
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB005352; green = RXRA, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB005352; green = RXRA, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RXRA IHC & IF
🧪Run bone marrow first and assess hematopoietic cells for RXRA staining (HPA: High in bone marrow hematopoietic cells). HPA detects RXRA in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and expect cells without specific signal on the positive slide to lack nuclear DAB staining above background without designating them RXRA-negative (HPA: no negative rows; standard IHC practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: RXRA 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 RXRA in A-431, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a secondary-only slide, a concentration-matched rabbit IgG isotype control, and RXRA-knockout tissue as a biological negative if available (caption: rabbit primary antibody; standard IHC practice). Block endogenous peroxidase and check for endogenous biotin signal in bone marrow when using the caption’s biotin-based DAB detection (caption: biotinylated secondary and SABC–DAB; standard IHC practice).
⚠️Feasibility: Paraffin-section IHC is documented with citrate retrieval at pH 6 for 20 minutes, but that caption does not establish whether retrieval is required for other preparations (caption: mouse liver IHC). The selected A01299-1 caption does not report a fixative, and no target-specific fixation window or fixation effect is reported in the supplied evidence (caption: fixative unreported). Frozen sections or IF/ICC cannot be judged easier from the supplied evidence; for bone marrow, endogenous peroxidase can complicate DAB interpretation (HPA: High in bone marrow hematopoietic cells; standard IHC practice).
HPA tissue IHC evidence for RXRA
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: RXRA is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced RXRA IHC Tips
Troubleshoot RXRA staining in paraffin sections by checking retrieval, nuclear localization, antibody specificity and cell-level scoring (datasheet A01299-1; HPA tissue IHC).
Which retrieval conditions should I start with for RXRA in paraffin sections?
Start with heat-mediated citrate buffer at pH 6 for 20 minutes (datasheet A01299-1). Keep heating, cooling and subsequent staining conditions consistent across sections while assessing whether nuclei become distinguishable from background (standard IHC practice; HPA tissue IHC: ubiquitous nuclear expression). The selected mouse liver image used paraffin sections and this retrieval method, but its caption does not state the fixative (datasheet A01299-1). If staining remains weak, compare a separately optimized retrieval condition on matched sections, recording tissue preservation and nuclear signal; do not treat a stronger chromogen deposit alone as evidence of improved specificity (standard IHC practice; HPA subcellular: nucleoplasm).
Could fixation explain weak or uneven RXRA staining?
Target-specific RXRA sensitivity to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not report its fixative (datasheet A01299-1). Record the actual fixative, fixation duration and processing history for each specimen before attributing weak nuclear staining to antigen loss (standard IHC practice; HPA tissue IHC: ubiquitous nuclear expression). Compare similarly processed sections using the same citrate pH 6, 20-minute retrieval and antibody conditions to isolate processing differences (datasheet A01299-1; standard IHC practice). Check nuclear morphology and staining at tissue edges before changing fixation conditions, because uneven processing can complicate interpretation (standard IHC practice).
How should I assess cytoplasmic RXRA staining alongside nuclear signal?
Prioritize a discernible nuclear pattern when assessing chromogenic RXRA staining, since tissue IHC reports ubiquitous nuclear expression and subcellular data place it mainly in the nucleoplasm (HPA tissue IHC; HPA subcellular). UniProt also lists cytoplasmic and mitochondrial localization, including translocation to mitochondria with NR4A1, so cytoplasmic staining is not automatically artefactual (UniProt P19793 subcellular; UniProt P19793 subunit). Compare nuclear and cytoplasmic compartments in intact cells at the same exposure and chromogen development settings (standard IHC practice). If only diffuse cytoplasm stains, inspect matched controls and morphology before assigning that deposit to RXRA (standard IHC practice; HPA subcellular: nucleoplasm).
Can this antibody distinguish RXRA isoforms or phosphorylation states in tissue?
The supplied record lists 2 RXRA isoforms, but gives no antibody epitope or isoform-specific reactivity for A01299-1 (UniProt P19793 isoforms; datasheet A01299-1). It also lists several phosphorylated residues, including positions 21, 27, 56, 70 and 82, without establishing how those modifications affect this antibody (UniProt P19793 modified residues). Treat chromogenic staining as antibody-associated RXRA signal, not a readout of a particular isoform or phosphorylation state (UniProt P19793 isoforms; standard IHC interpretation). If that distinction matters, obtain documented epitope and reactivity information and validate the intended comparison with appropriate controls before interpreting tissue differences (standard IHC practice).
How can I compare RXRA with a cell-type marker by immunofluorescence?
On a separate IF/ICC workflow, pair RXRA with a validated marker for the cell type being examined and assess overlap within segmented cells (standard IF practice). Choose spectrally separated fluorophores and inspect unstained tissue for autofluorescence before assigning a weak RXRA signal, particularly in the channel selected for RXRA (standard IF practice). RXRA has no transmembrane segment and is mainly nucleoplasmic, so use permeabilisation suitable for access to an intracellular nuclear epitope while preserving nuclear morphology (UniProt P19793 topology; HPA subcellular; standard IF practice). HPA lists A-431 and U2OS ICC/IF images, but the supplied evidence does not establish an IF protocol or IF validation for A01299-1 (HPA subcellular; datasheet A01299-1).
What should I check when RXRA DAB staining is diffuse?
Separate diffuse deposit from cell-associated signal by inspecting nuclei, blank areas and section edges at the same microscope settings (standard IHC practice; HPA tissue IHC: ubiquitous nuclear expression). The selected caption used 10% goat serum, 1 μg/ml primary antibody overnight at 4°C, a biotinylated secondary, an avidin–biotin complex and DAB (datasheet A01299-1). Review primary omission and reagent controls, peroxidase blocking, secondary binding and DAB development when deposit appears without convincing nuclear structure (standard chromogenic IHC practice). Because that detection scheme uses biotin, evaluate endogenous biotin as a possible contributor to background in the specimen being tested (datasheet A01299-1; standard IHC practice).
How should I score RXRA staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and nuclear scoring rules before comparing slides, since the reported tissue pattern is broadly nuclear (HPA tissue IHC: ubiquitous nuclear expression; standard IHC practice). Record the percentage of positive nuclei and, where intensity is reproducible, a nuclear H-score using fixed intensity categories; report cytoplasmic staining separately (standard IHC practice; UniProt P19793 subcellular). Normalize positive-cell counts or density per mm² to the evaluated viable tissue area and report the number of eligible cells (standard IHC practice). Keep retrieval, detection, counterstain, imaging and thresholds consistent across groups, and exclude damaged edges or necrotic regions by a stated rule (standard IHC practice).
When is an apparent RXRA-positive cell likely an artefact?
A true-positive call is more convincing when chromogen follows intact nuclei in an identifiable cell population, consistent with the reported ubiquitous nuclear tissue pattern (HPA tissue IHC; standard IHC interpretation). HPA reports high staining in adrenal glandular cells, bone-marrow hematopoietic cells and bronchial respiratory epithelial cells, but its tissue reliability remains Supported with external verification pending (HPA tissue IHC). Question signal confined to torn edges, necrotic areas or extracellular material, and check peroxidase and primary-omission controls for reagent-derived deposit (standard IHC practice). Cytoplasmic signal warrants separate assessment because UniProt also reports cytoplasmic and mitochondrial RXRA; compartment alone cannot establish specificity (UniProt P19793 subcellular; standard IHC interpretation).
Boster reagents
Best RXRA / Retinoic acid receptor RXR-alpha IHC Antibodies
The IHC-validated RXRA antibody has paraffin-section images from human, mouse and rat tissues (A01299-1 IHC captions); IF imaging covers A431 cells (A01299-1 IF caption).
Real IHC data
Anti-Retinoid X Receptor alpha/RXRA Antibody ®
Cat # A01299-1
A01299-1 will render with its own IHC figure from a mouse liver paraffin section (A01299-1 IHC figure caption). Its catalog also documents IHC in rat liver, human liver and human liver cancer tissue, plus IF in A431 cells (A01299-1 image captions).
Which to pick: For tissue IHC, choose A01299-1: its paraffin-section captions document human, mouse and rat samples; the fixative is unreported (A01299-1 IHC captions). For IF/ICC, A01299-1 has an A431 cell image, while M01299-2 is a rabbit monoclonal listed for IF/ICC without an accompanying image (A01299-1 IF caption; M01299-2 catalog). For cross-species tissue IHC, A01299-1 has image evidence across all three listed species (A01299-1 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P19793 (RXRA_HUMAN, Retinoic acid receptor RXR-alpha).
- Human Protein Atlas. RXRA tissue IHC expression (reliability: Supported).
- Human Protein Atlas. RXRA subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the Golgi apparatus..
- Human Protein Atlas. RXRA antibody validation summary (2 antibodies).
- TEAD4 and RXRA Regulate the Function of Nucleus Pulposus Cells in Intervertebral Disc Degeneration Via the TNF-α/NF-κB Pathway: An Integrated Analysis of Single-Cell RNA-Seq, Bulk RNA-Seq, and In Vitro Validation. Applied biochemistry and biotechnology 2026 — PMC12995935.
- Gaseous Air Pollutant Exposure and Atherosclerosis: A Systematic Study Integrating Multi-Omics and Network Toxicology. International journal of molecular sciences 2026 — PMC13513811.
- PubMed PMID:2159111 — UniProt-cited evidence.
- PubMed PMID:18619963 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RXRA (Retinoic acid receptor RXR-alpha). Expected localisation Ubiquitous nuclear tissue staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A01299-1). Validated IHC antibody: A01299-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular 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 mouse liver tissue; fixative not specified (datasheet A01299-1); 2. Fixation: Image fixative and duration unreported (datasheet A01299-1); 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: Citrate pH 6 , 20 min (datasheet A01299-1); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01299-1); 8. Primary antibody: Rabbit anti-RXRA, 0.5-1μg/ml (datasheet A01299-1); 9. Primary incubation: Overnight at 4 °C (datasheet A01299-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01299-1); Caveat: Widespread nuclear staining limits tissue specificity (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences are unspecified (UniProt); Regulation: Staining-intensity regulation is unreported (UniProt). Evidence: UniProt P19793, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A01299-1, PMC12995935. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RXRA/immunohistochemistry-ihc-guide.