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RASD2 / GTP-binding protein Rhes · IHC design guide
Design Immunohistochemistry for RASD2
Plan chromogenic RASD2 IHC-P around the cytoplasmic tissue pattern (HPA tissue IHC). Start the catalog antibody at 2.5 μg/mL (datasheet: IHC-P), using seminal vesicle glandular cells as a high-staining reference and salivary gland glandular cells as a not-detected comparator (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RASD2 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 | Cytoplasmic tissue staining (HPA tissue IHC); cell membrane is a molecular location (UniProt) | |
| Staining pattern | Cytoplasmic staining in glandular cells across several tissues (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Seminal vesicle+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); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A06806) | |
| Caveat | Brain-enriched RNA may not predict IHC intensity (HPA tissue IHC) | |
| Regulation | Staining-linked regulation is not specified (UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain spans residues 1–263 (UniProt) |
Section 1
Recommended RASD2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by published RASD2 IHC methods for renal carcinoma (PMC13394055), thyroid cancer (PMC12199591), and mouse brain (PMC11746134).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A06806); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-RASD2, 2.5 μg/mL (datasheet A06806) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RASD2-positive staining in glandular cells of seminal vesicle (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteFor paraffin sections, start with citrate pH 6.0 at 95–98 °C for 20 min (page retrieval setting); renal tissue has a published pH 9.0 alternative (PMC13394055).
Section 2
What Is the Expected RASD2 Staining Pattern?
RASD2 is annotated at the cell membrane without a transmembrane segment (UniProt Q96D21 topology). In tissue IHC, expect predominantly cytoplasmic staining in selected cells, including seminal vesicle glandular cells and several glandular or epithelial populations (HPA tissue IHC). HPA rates the tissue pattern “Enhanced,” while reporting medium consistency with RNA data and pending external verification (HPA tissue IHC).
| Strong chromogenic staining in seminal vesicle glandular cells, with weaker staining in nearby cells (HPA: High in seminal vesicle glandular cells). | This matches the clearest listed tissue positive. Assess the stained cell population and its cytoplasmic pattern together; intensity alone cannot establish antibody specificity (HPA tissue IHC; general IHC practice). |
| Predominantly isolated nuclear staining, without a convincing cytoplasmic pattern. | Treat this as a compartment mismatch and investigate artefact. UniProt places RASD2 at the cell membrane, and HPA describes cytoplasmic tissue expression; neither source supports a nuclear IHC pattern (UniProt Q96D21; HPA tissue IHC). |
| Prominent staining in a cell population listed as undetected, such as adipocytes or salivary gland glandular cells (HPA tissue IHC). | Consider cross-reactivity or endogenous detection activity before calling these cells RASD2 positive. HPA’s negative calls apply to the named cell populations, not necessarily to every cell in those tissues (HPA tissue IHC; general IHC practice). |
| Widespread, poorly bounded color across cells and surrounding tissue, obscuring cell-specific staining. | Interpret the field as diffuse background until controls clarify it. Compare with a control lacking primary antibody and review blocking, washing, and detection conditions (general IHC practice); HPA reports a cell-associated tissue pattern (HPA tissue IHC). |
| No convincing staining in seminal vesicle glandular cells (HPA: High in seminal vesicle glandular cells). | An absent signal in this listed positive limits interpretation of other negatives. Check tissue preservation, antibody and detection controls, and the assay’s retrieval conditions before concluding that RASD2 is absent (HPA tissue IHC; general IHC practice). |
💡Expected RASD2 appearanceCall a result consistent with RASD2 when seminal vesicle glandular cells show high, predominantly cytoplasmic staining (HPA tissue IHC); isolated nuclear color or staining confined to an HPA-listed undetected cell population is suspect (UniProt Q96D21; HPA tissue IHC).
| Compartment and topology | UniProt assigns RASD2 to the cell membrane and lists no transmembrane segment; HPA tissue IHC describes cytoplasmic expression. Evaluate tissue staining against both annotations without requiring a sharp membrane rim in every positive cell (UniProt Q96D21 topology; HPA tissue IHC). |
| Tissue and cell selection | HPA lists high staining in seminal vesicle glandular cells, medium staining in several other named populations, and low staining in caudate neuronal cells. Choose and score the named cells rather than treating an entire tissue as uniformly positive (HPA tissue IHC). |
| Strength of the tissue evidence | HPA assigns “Enhanced” tissue IHC reliability and lists HPA005839 as IHC Enhanced, while describing medium agreement with RNA data and pending external verification. Use the reported pattern as a reference, with controls for the specimen under study (HPA tissue IHC; HPA antibodies). |
| Processing and epitope interpretation | UniProt annotates a chain spanning residues 1–263, a propeptide at 264–266, and a modified cysteine at residue 263. The payload gives no antibody epitope, so it cannot predict how these features affect this assay’s staining (UniProt Q96D21 processing). |
| Can IF/ICC show the same localisation? | HPA reports approved plasma membrane and cytosol localisation in ICC-IF, with images from U2OS cells; HPA005839 is ICC Approved. This supports a localisation comparison, while the tissue intensity calls on this page come from IHC (HPA subcellular; HPA antibodies; HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive tissue is blank. | The assay may have failed, or the section may lack evaluable seminal vesicle glandular cells; HPA reports those cells as high (HPA tissue IHC). | Verify cell identity and section quality, then review the antibody, retrieval, and detection controls used for this IHC run (general IHC practice). |
| Only nuclei are strongly colored. | The pattern conflicts with membrane annotation and cytoplasmic tissue staining; nonspecific staining or a detection artefact is possible (UniProt Q96D21; HPA tissue IHC). | Compare a control lacking primary antibody, inspect counterstain and tissue pigment, and require a plausible cytoplasmic cell pattern before scoring (general IHC practice). |
| An HPA-listed undetected cell population stains strongly. | Cross-reactivity or endogenous detection activity is possible; HPA lists adipocytes and salivary gland glandular cells as undetected (HPA tissue IHC; general IHC practice). | Identify the exact stained cells and compare controls lacking primary antibody. Resolve any background before interpreting the unexpected signal as RASD2 (general IHC practice). |
| Most of the section has diffuse color. | Background from blocking, washing, or detection can obscure the cell-associated pattern reported by HPA (general IHC practice; HPA tissue IHC). | Review blocking and washes, inspect a control lacking primary antibody, and score only cells whose signal remains distinguishable from background (general IHC practice). |
| Caudate neurons stain weakly despite brain-enriched RNA. | HPA lists caudate neuronal staining as low; its brain-enriched RNA category does not specify high protein staining in those cells (HPA tissue IHC). | Use the cell-specific IHC observation when judging the slide, and compare a listed stronger positive before treating weak caudate staining as assay failure (HPA tissue IHC; general IHC practice). |
| An ICC-IF image appears more membrane-defined than the tissue IHC slide. | HPA approves plasma membrane and cytosol localisation in ICC-IF, while its tissue IHC summary describes cytoplasmic expression (HPA subcellular; HPA tissue IHC). | Judge the chromogenic section against the tissue IHC cell pattern. Use ICC-IF only as supporting localisation evidence, without transferring its appearance directly to tissue scoring (HPA tissue IHC; HPA subcellular). |
Experiment evidence
IHC & IF images showing RASD2 staining
A06806 · IHCImmunohistochemistry of TEM2 in human colon tissue with TEM2 antibody at 2.5 μg/mL.
A06806 · IFImmunofluorescence of TEM2 in Human Colon cells with TEM2 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000100302-RASD2/tissue/cerebral+cortex#imid_1871900
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000100302-RASD2/tissue/colon#imid_1871834
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000100302-RASD2/tissue/liver#imid_1871835
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000100302-RASD2/tissue/kidney#imid_1871820
HPA · ICC-IF U2OSHuman Protein Atlas immunofluorescence, U2OS, antibody HPA005839; green = RASD2, blue = nucleus, red = microtubules.
HPA · ICC-IF U2OSHuman Protein Atlas immunofluorescence, U2OS, antibody HPA005839; green = RASD2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RASD2 IHC & IF
🧪Run seminal vesicle first; its glandular cells should stain (HPA: High in seminal vesicle glandular cells). Use adipose tissue as a negative comparator (HPA: Not detected in adipocytes), and treat nonglandular cells on the positive slide as internal negative comparators only where they lack staining above background.
Positive control tissue: Seminal vesicle (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RASD2 in U2OS, with annotated localisation: Plasma membrane (approved), Cytosol (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and clonality-matched isotype controls; use RASD2 knockout material or a cognate peptide block, if available, to assess specificity. Quench endogenous peroxidase for chromogenic IHC, and assess background in seminal-vesicle luminal material separately from glandular-cell staining (HPA: High in seminal vesicle glandular cells).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence; the selected A06806 tissue-IHC caption does not state a fixative (selected A06806 caption: fixative not stated). Antigen-retrieval dependence is unreported, and the supplied evidence does not establish whether frozen sections or IF are easier than paraffin IHC. For seminal vesicle, score glandular cells separately from luminal material (HPA: High in seminal vesicle glandular cells).
HPA tissue IHC evidence for RASD2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Seminal vesicle | Glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuropil | Medium | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RASD2 IHC Tips
Troubleshoot RASD2 chromogenic IHC in paraffin sections using the page retrieval setting, the catalog tissue image, and compartment and cell type controls.
How should I adjust retrieval when RASD2 staining is weak or uneven?
Start with citrate pH 6.0 heat retrieval at 95–98 °C for 20 min (page antigen retrieval setting). If staining is weak, first check that sections remained covered by buffer and that the vessel reached the specified temperature; compare runs using the same heating and cooling procedure. Then compare a shorter and a longer retrieval time on adjacent sections while keeping antibody concentration and chromogenic development constant (general IHC practice). Examine tissue integrity and background alongside signal, because excessive heating can damage morphology or increase nonspecific staining (general IHC practice). Score the expected glandular compartment in colon rather than treating stronger stain anywhere as improvement (HPA: medium in colon glandular cells).
Can fixation explain a weak RASD2 signal in paraffin sections?
Target-specific fixation sensitivity for RASD2 is unknown from the supplied evidence; the catalog colon image gives no fixative (A06806 tissue-IHC caption: fixative not stated). Record each specimen’s fixative, fixation duration, section thickness, and processing history before comparing staining across blocks (general IHC practice). For a practical comparison, stain adjacent sections from differently processed blocks in one run with the same pH 6.0 retrieval and antibody conditions (page antigen retrieval setting; general IHC practice). Include a tissue and cell compartment expected to stain, such as colon glandular cells, to distinguish a weak run from a weak specimen (HPA: medium in colon glandular cells). Do not assign a fixation mechanism without a controlled comparison.
Is cytoplasmic RASD2 staining credible when I expect membrane localisation?
Interpret a cytoplasmic pattern in context: tissue IHC reports cytoplasmic expression, while cell imaging places RASD2 at the plasma membrane and in the cytosol (HPA tissue IHC: cytoplasmic expression; HPA subcellular: approved plasma membrane and cytosol). Its annotated location is the cell membrane, with no transmembrane segment and a modified cysteine near the processed C terminus (UniProt Q96D21: subcellular location, topology, processing, modified residue). On serial sections, compare staining within cells against morphology and the distribution of the expected cell type (general IHC practice). Diffuse signal across every compartment or a sharp section-edge gradient warrants review of blocking, development, and section handling before it is called localisation (general IHC practice).
How can I troubleshoot an epitope-dependent staining discrepancy?
Check the antibody’s stated immunogen or epitope against the processed RASD2 chain, amino acids 1–263, before interpreting a negative paraffin section (UniProt Q96D21: processing). The supplied record annotates 0 isoforms, 0 glycosylation sites, and a cysteine methyl ester at residue 263 (UniProt Q96D21: isoforms, glycosylation, modified residue). These annotations alone do not establish whether fixation, retrieval, or the modification masks this antibody’s epitope. Compare antibody concentrations and the page’s citrate pH 6.0 retrieval on adjacent sections, holding detection constant (page antigen retrieval setting; general IHC practice). If epitope information is unavailable, report that limitation and use a separately validated antibody or orthogonal expression evidence to investigate disagreement (general IHC practice).
What should I consider before checking the IHC pattern by IF?
Treat IF as a separate validation experiment: the supplied catalog image documents tissue IHC, while the subcellular IF record reports imaging in U2OS cells (A06806 tissue-IHC caption; HPA subcellular: U2OS). For tissue multiplexing, pair RASD2 with a validated marker identifying the expected glandular cells, then ask whether signals occupy the same cells rather than merely overlapping adjacent structures (HPA: medium in colon glandular cells; general IF practice). Choose spectrally separated fluorophores after inspecting unstained tissue for autofluorescence, and include single-label controls to assess bleed-through (general IF practice). Test permeabilised and minimally permeabilised conditions when epitope accessibility is uncertain, because RASD2 is reported in cytosol and at the plasma membrane, but the antibody’s epitope side is unspecified (HPA subcellular: approved cytosol and plasma membrane; general IF practice).
How do I reduce diffuse or granular background in RASD2 IHC?
Compare a no-primary control with the stained section to locate signal arising from secondary detection, endogenous peroxidase, or chromogen deposition (general chromogenic IHC practice). Apply a peroxidase block before horseradish peroxidase detection, use an appropriate protein block, and keep DAB development time consistent across sections (general chromogenic IHC practice). If diffuse staining remains, titrate the catalog antibody below the 2.5 µg/mL concentration shown in its colon image while holding retrieval and exposure fixed (A06806 tissue-IHC caption; general IHC practice). Judge improvement in the expected glandular cells and against a no-primary control, since colon glandular staining is reported but a uniformly stained section is not a useful specificity check (HPA: medium in colon glandular cells; general IHC practice).
What is a defensible way to quantify RASD2 chromogenic staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the region and cell population before scoring, then record the percentage of positive cells and an H-score from intensity categories 0–3 in the same compartment (general IHC scoring practice). For areas where individual cells cannot be resolved reliably, report positive area or optical density per mm² of viable tissue and state the thresholding method (general image analysis practice). Normalise cell counts to the number of evaluable cells, or area measures to viable tissue area, using identical acquisition and DAB settings across groups (general IHC practice). Keep glandular cells and neuropil in separate analyses because both have reported staining but are different measurement units (HPA: medium in colon glandular cells and cerebral cortex neuropil).
How can I distinguish a true RASD2 signal from artefact?
A plausible signal should fit the examined tissue and compartment: colon glandular cells have reported staining, and RASD2 is reported in cytosol and at the plasma membrane (HPA: medium in colon glandular cells; HPA subcellular: approved cytosol and plasma membrane). Compare the same run with a no-primary control and inspect section edges, folds, and necrotic areas before assigning positive cells (general IHC practice). Staining confined to damaged tissue or matching residual endogenous enzyme signal calls for technical investigation, including the peroxidase block, rather than a biological interpretation (general chromogenic IHC practice). Treat a mismatch with expected cell type or localisation cautiously because the tissue IHC profile has only medium staining–RNA consistency and awaits external verification (HPA tissue IHC: Enhanced reliability description).
Boster reagents
Best RASD2 / GTP-binding protein Rhes IHC Antibodies
A06806 has IHC and IF images from human colon samples (catalog image captions). Its listed reactivity covers human, mouse, and rat (catalog reactivity).
Real IHC data
Anti-TEM2 RASD2 Antibody
Cat # A06806
A06806 will render with an IHC image of human colon tissue at 2.5 μg/mL (A06806 IHC image caption). The same SKU lists IHC-P and IF applications and has an IF image of human colon cells at 20 μg/mL (A06806 applications; A06806 IF image caption).
Which to pick: For paraffin-section IHC, choose A06806: it lists IHC-P and shows human colon tissue staining; the image caption does not report a fixative (A06806 applications; A06806 IHC image caption). For IF/ICC, A06806 has an IF image of human colon cells, but ICC-specific validation is unreported (A06806 IF image caption; A06806 applications). For work across species, A06806 lists human, mouse, and rat reactivity; the supplied IHC and IF images show human samples only (A06806 reactivity; A06806 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96D21 (RHES_HUMAN, GTP-binding protein Rhes).
- Human Protein Atlas. RASD2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. RASD2 subcellular location (ICC-IF): Localized to the plasma membrane and cytosol..
- Human Protein Atlas. RASD2 antibody validation summary (1 antibodies).
- RASD2 Drives Renal Clear Cell Carcinoma Progression via RAF1 (Ser338) Phosphorylation. Cancer science 2026 — PMC13394055.
- Modulation of RASD2 by miRNA-485-5p Drives Thyroid Cancer Progression and Metastasis. The Kaohsiung journal of medical sciences 2025 — PMC12199591.
- Rasd2 regulates depression-like behaviors via DRD2 neurons in the prelimbic cortex afferent to nucleus accumbens core circuit. Molecular psychiatry 2025 — PMC11746134.
- PubMed PMID:10947988 — UniProt-cited evidence.
- PubMed PMID:15461802 — UniProt-cited evidence.
- PubMed PMID:10591208 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RASD2 (GTP-binding protein Rhes). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); cell membrane is a molecular location (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A06806 at 2.5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Seminal vesicle (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A06806); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-RASD2, 2.5 μg/mL (datasheet A06806); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A06806); Caveat: Brain-enriched RNA may not predict IHC intensity (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain spans residues 1–263 (UniProt); Regulation: Staining-linked regulation is not specified (UniProt). Evidence: UniProt Q96D21, HPA tissue IHC · Enhanced, HPA ICC-IF · plasma membrane and cytosol., Datasheet A06806, PMC13394055, PMC12199591. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RASD2/immunohistochemistry-ihc-guide.