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RHOG / Rho-related GTP-binding protein RhoG · IHC design guide
Design Immunohistochemistry for RHOG
Plan RHOG chromogenic IHC around cytoplasmic staining in a subset of immune cells (HPA tissue IHC). Bone marrow hematopoietic cells show high staining (HPA tissue IHC); the catalog antibody starts at 5 μg/mL, with IHC validation reported in mouse tissue (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RHOG 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 staining in immune cells (HPA tissue IHC); membrane localization at the molecular level (UniProt) | |
| Staining pattern | A subset of immune cells shows cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Bone marrow+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 | Staining varies across immune-cell subsets (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | No isoforms; mature chain spans residues 1–188 (UniProt) |
Section 1
Recommended RHOG IHC & IF Protocols
The catalog antibody protocol is followed by one published RHOG IHC protocol for paraffin sections (PMC4232683: Methods).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A03485); 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-RHOG, 5 μg/mL (datasheet A03485) |
| 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 | RHOG-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in a subset of immune cells. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 heat retrieval: it matches the page’s retrieval setting and the published protocol (page antigen retrieval; PMC4232683: Methods).
Section 2
What Is the Expected RHOG Staining Pattern?
RHOG staining in paraffin sections should be cytoplasmic in a subset of immune cells, with possible peripheral accentuation because UniProt places RHOG at the cell membrane without a transmembrane segment (HPA: tissue IHC profile; UniProt P84095: subcellular location and topology). HPA rates its tissue IHC evidence Enhanced, while describing only medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability).
| Strong cytoplasmic staining in bone marrow hematopoietic cells or tonsil germinal center cells. | This fits two high staining compartments; score the named cells rather than treating every cell in the section as positive (HPA: High in both compartments). |
| Predominantly nuclear staining in cells otherwise expected to be positive. | This conflicts with the reported cytoplasmic IHC pattern and cell membrane annotation; check whether the signal follows nuclei in a no-primary control (HPA: tissue IHC profile; UniProt P84095: cell membrane; standard IHC practice). |
| Strong staining in bronchial respiratory epithelium or esophageal squamous epithelium. | HPA reports RHOG as undetected in those cell types; investigate cross-reactivity or endogenous detection activity before calling the signal RHOG (HPA: Not detected in both compartments; standard IHC practice). |
| Color spreads across tissue and empty areas without clear cell boundaries. | This is inconsistent with a selected-cell cytoplasmic pattern; assess nonspecific staining and detection background with appropriate controls (HPA: tissue IHC profile; standard IHC practice). |
| No staining in bone marrow hematopoietic cells on the test section. | A negative result there conflicts with HPA's high staining report; verify tissue preservation, retrieval, primary antibody and detection performance before interpreting another section as RHOG-negative (HPA: High in bone marrow hematopoietic cells; standard IHC practice). |
💡Expected RHOG appearanceCall a result positive when selected bone marrow hematopoietic cells show strong cytoplasmic staining, possibly accentuated at the edge; isolated strong epithelial or nuclear staining warrants investigation (HPA: High in bone marrow hematopoietic cells and cytoplasmic immune-cell profile; UniProt P84095: cell membrane; standard IHC practice).
| Choice of tissue compartment | Bone marrow hematopoietic cells, splenic red pulp cells and tonsil germinal center cells are high staining examples; lymph node non-germinal center cells are medium (HPA: tissue IHC levels). |
| Antibody evidence | HPA039871 is a rabbit polyclonal antibody with Enhanced IHC validation, meaning its pattern was supported by independent antibodies or orthogonal data; this does not establish every new run's specificity (HPA: antibody validation; standard IHC practice). |
| Localization and topology | UniProt assigns RHOG to the cell membrane and lists no transmembrane segment; HPA describes cytoplasmic staining in a subset of immune cells, so a crisp membrane rim is not required for a positive IHC call (UniProt P84095: location and topology; HPA: tissue IHC profile). |
| Processing and epitope position | UniProt lists a chain at residues 1–188 and a propeptide at 189–191; without an antibody epitope map, those annotations cannot predict which compartment will stain or whether processing affects detection (UniProt P84095: processing). |
| Endogenous detection activity | Background from endogenous detection activity can mimic specific chromogenic signal; a no-primary control helps assess it, especially when unexpected cells stain (standard IHC practice). |
| IF/ICC: is the IHC pattern an IF localization reference? | Use the membrane annotation as biological context, but HPA supplies no main ICC-IF location or cell-line images here; it does not provide an IF pattern to validate against (UniProt P84095: cell membrane; HPA: subcellular record). |
| Situation | Likely cause | Next action |
|---|---|---|
| Bone marrow hematopoietic cells are blank. | The expected high staining compartment is absent, or the IHC detection workflow has failed (HPA: High in bone marrow hematopoietic cells; standard IHC practice). | Confirm the annotated cell population, then review the run's retrieval, primary antibody and detection controls before scoring a negative result (standard IHC practice). |
| Tonsil germinal center staining is weak. | The observed intensity falls below HPA's high category, though intensity can vary between IHC runs (HPA: High in tonsil germinal center cells; standard IHC practice). | Check staining of a known positive compartment in the same run and review the antibody's documented IHC conditions before changing interpretation (HPA: high staining compartments; standard IHC practice). |
| Bronchial respiratory epithelium stains strongly. | That cell type is reported as undetected, raising concern for nonspecific primary staining or endogenous detection activity (HPA: Not detected in bronchial respiratory epithelial cells; standard IHC practice). | Compare a no-primary control and the tissue's expected immune-cell staining; investigate persistent epithelial signal before assigning it to RHOG (HPA: cytoplasmic immune-cell profile; standard IHC practice). |
| Nuclei dominate the signal. | Nuclear dominance conflicts with the reported cytoplasmic IHC profile and membrane annotation (HPA: tissue IHC profile; UniProt P84095: cell membrane). | Inspect the counterstain and no-primary control, then review antibody specificity and detection background (standard IHC practice). |
| Diffuse color obscures cell boundaries. | The section cannot be assessed against HPA's subset-specific cytoplasmic pattern while background dominates (HPA: tissue IHC profile; standard IHC practice). | Check no-primary and detection controls, then optimize routine blocking or wash conditions as indicated by those controls (standard IHC practice). |
| Staining varies sharply among immune-cell groups. | HPA reports high staining in some named compartments and medium staining in lymph node non-germinal center cells; mixed-cell averages can hide that difference (HPA: tissue IHC levels). | Identify and score the annotated cell populations separately, keeping the HPA compartment labels with each comparison (HPA: tissue IHC levels; standard IHC practice). |
Experiment evidence
IHC & IF images showing RHOG staining
A03485 · IHCImmunohistochemistry of RHOG in mouse kidney tissue with RHOG antibody at 5 μg/ml.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000177105-RHOG/tissue/cerebral+cortex#imid_10272333
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000177105-RHOG/tissue/colon#imid_10272243
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000177105-RHOG/tissue/liver#imid_10272252
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000177105-RHOG/tissue/kidney#imid_10272245
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RHOG IHC & IF
🧪Run bone marrow first and expect hematopoietic cells to stain (HPA: High in bone marrow hematopoietic cells). Use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipocytes); on the marrow slide, assess background in cells outside the listed hematopoietic population without assuming they are RHOG-negative.
Positive control tissue: Bone marrow (Hematopoietic cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for RHOG; derive a cell-line control from the positive tissue's cell type (Hematopoietic cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; an irrelevant antibody matched to the primary antibody’s host species, isotype, and clonality; and RHOG knockout tissue or a peptide-block control if the immunizing peptide is available (standard IHC practice). Quench endogenous peroxidase in bone marrow before chromogenic detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected mouse-kidney IHC caption does not state a fixative (A03485 IHC caption: fixative not stated). Antigen-retrieval dependence is unreported; optimize retrieval on matched paraffin sections and watch for marrow peroxidase background (standard IHC practice). The supplied evidence does not establish whether frozen sections or IF are easier; HPA lists no cell lines with RHOG ICC-IF images (HPA subcellular: none).
HPA tissue IHC evidence for RHOG
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Skin | Fibrohistiocytic cells | High | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | High | Protein (IHC) | HPA → |
| Tonsil | Germinal center cells | High | Protein (IHC) | HPA → |
| Lymph node | Non-germinal center cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Colon | Endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RHOG IHC Tips
Troubleshoot RHOG staining in paraffin sections by checking retrieval, cell identity, staining compartment, and controls before comparing signal between samples.
How should I optimize retrieval when RHOG staining is weak in paraffin sections?
Use citrate pH 6.0 HIER at 95–98 °C for 20 min as the starting retrieval condition for RHOG paraffin IHC (page retrieval rule). Compare retrieved and unretrieved sections from the same block, keeping antibody concentration and chromogen development constant so that changes in staining can be attributed to retrieval (standard IHC practice). Assess signal in identifiable immune cell populations alongside tissue morphology, since RHOG staining is reported in a subset of immune cells (HPA: tissue IHC profile). If signal remains weak, test a more intensive retrieval condition as a fallback on matched sections and reject conditions that increase background or damage morphology (standard IHC practice).
Could fixation explain weak or uneven RHOG staining in my paraffin sections?
RHOG-specific sensitivity to fixation is unknown from the supplied evidence; record the fixative and fixation history for every block before attributing weak staining to fixation. The catalog antibody has a mouse kidney tissue-IHC image at 5 µg/ml, but its caption does not state the fixative (selected A03485 tissue-IHC caption). Compare sections with documented processing histories under the same retrieval, antibody incubation, and chromogen development conditions to identify a processing-associated difference (standard IHC practice). Include a tissue with identifiable hematopoietic cells as a biological reference, while recognizing that their high staining in HPA does not establish a fixation protocol (HPA: High in bone marrow hematopoietic cells).
Should RHOG staining be cytoplasmic, membranous, or nuclear in IHC?
Evaluate RHOG primarily as cytoplasmic staining with possible membrane-associated accentuation, rather than using isolated nuclear staining as the expected pattern (HPA: cytoplasmic expression in a subset of immune cells; UniProt P84095: cell membrane). RHOG has no transmembrane segment, and activation-dependent phospholipid binding provides a basis for membrane association without implying a continuous membrane rim in every fixed cell (UniProt P84095: topology and function). Examine cell borders and cytoplasm at matched magnification, recording each compartment separately rather than combining them into one score (standard IHC practice). Treat a nuclear-only pattern as requiring independent validation, particularly if the expected immune cell population is unstained (HPA: tissue IHC profile; standard IHC practice).
How do epitope placement and RHOG processing affect interpretation of IHC staining?
Check the antibody's documented immunogen or epitope before interpreting discordant RHOG stains; the supplied record does not specify an epitope for the catalog antibody. The record lists 0 isoforms and a mature chain spanning residues 1–188, with a propeptide at 189–191 (UniProt P84095: isoforms and processing). It also annotates modifications at residues 39, 138, 180, and 188, but their effects on this antibody's paraffin-section binding are untested here (UniProt P84095: modified residues). If antibodies give different compartment patterns, compare their documented epitopes and staining in the same identified cell population before assigning a biological explanation (standard IHC practice).
How can IF help verify a questionable RHOG IHC pattern?
Use IF on a matched specimen to examine RHOG alongside a validated marker identifying the immune cell population of interest; the expected tissue-IHC pattern involves a subset of immune cells (HPA: tissue IHC profile). Select spectrally separated fluorophores and assess unstained tissue in each channel, because tissue autofluorescence can mimic a weak cellular signal (standard IF practice). Choose permeabilisation based on the documented antibody epitope: an intracellular-facing epitope needs access through the plasma membrane, while RHOG itself has no transmembrane segment (UniProt P84095: topology; standard IF practice). Compare IF localisation with IHC cautiously, since no RHOG ICC/IF image or IF-specific fixation condition is supplied here (HPA: subcellular record).
What controls distinguish RHOG staining from diffuse chromogenic background?
Run a no-primary control through the same detection and chromogen steps, then inspect whether diffuse color persists in the cells and compartments being scored (standard IHC practice). Apply the routine endogenous-peroxidase block for a peroxidase-based assay, and compare DAB development times on matched sections so that excess reaction product is not mistaken for RHOG (standard IHC practice). Inspect section edges, folds, damaged areas, and pigment separately from intact cells, since these regions can complicate chromogenic interpretation (standard IHC practice). Use identified hematopoietic cells as a positive biological reference and adipocytes as a reported negative cell type, while accounting for specimen-specific variation (HPA: High in bone marrow hematopoietic cells; Not detected in adipocytes).
How should I quantify RHOG staining across paraffin-section specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because HPA reports cytoplasmic staining in a subset of immune cells (HPA: tissue IHC profile). Within the same population, report the percentage of positive cells and, when intensity categories are reproducible, an H-score calculated as the sum of each intensity score multiplied by its percentage of cells (standard IHC scoring practice). For infiltrates, also report positive-cell density per mm² of evaluable tissue and normalise comparisons to the relevant cell population or tissue area (standard IHC quantification practice). Hold retrieval, antibody concentration, imaging settings, and DAB development constant across specimens; document exclusions such as folds and necrotic regions (standard IHC practice).
When is a positive RHOG IHC result biologically convincing?
A convincing result places reproducible cytoplasmic or membrane-associated signal in identifiable cells, especially immune cell populations consistent with the reported tissue pattern (HPA: tissue IHC profile; UniProt P84095: cell membrane). High staining is reported in bone marrow hematopoietic cells, whereas adipocytes are reported as not detected; use these as interpretive references, not absolute specimen-level controls (HPA: tissue IHC). Question a nuclear-only signal, uniform staining across unrelated cell types, or color restricted to cut edges and necrotic areas, and compare each finding with the no-primary section (UniProt P84095: localisation; standard IHC practice). If signal survives those checks, describe the stained cell population and compartment rather than inferring RHOG activity from protein staining alone (standard IHC interpretation practice).
Boster reagents
Best RHOG / Rho-related GTP-binding protein RhoG IHC Antibodies
A03485 has real RHOG IHC data from mouse kidney tissue (image caption: mouse kidney). Its listed reactivity covers human, mouse and rat; the pictured IHC result is from mouse tissue (catalog: reactivity; image caption).
Real IHC data
Anti-RHOG Antibody
Cat # A03485
A03485 will render with an IHC image of mouse kidney tissue stained at 5 μg/mL (image caption). It lists IHC-P and human, mouse and rat reactivity, while the catalog specifies IHC validation in mouse samples (catalog: applications, reactivity and validation).
Which to pick: Choose A03485 for paraffin-section IHC: IHC-P is listed, and its own image shows mouse kidney staining at 5 μg/mL (catalog: IHC-P; image caption: mouse kidney, 5 μg/mL). No IF/ICC application or image is supplied for A03485, so there is no IF/ICC pick here; the fixative used for the pictured tissue is unreported (catalog: applications and images; image caption). For work across species, A03485 is the only listed choice and is described as reactive with human, mouse and rat, but its stated IHC validation is in mouse samples (catalog: reactivity and validation).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P84095 (RHOG_HUMAN, Rho-related GTP-binding protein RhoG).
- Human Protein Atlas. RHOG tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. RHOG subcellular location (ICC-IF): Highest expression in HEL: 215.5 nTPM.
- Human Protein Atlas. RHOG antibody validation summary (1 antibodies).
- The small GTPase RhoG mediates glioblastoma cell invasion. Molecular cancer 2012 — PMC3557187.
- Host response profile of human brain proteome in toxoplasma encephalitis co-infected with HIV. Clinical proteomics 2014 — PMC4232683.
- SGEF is a potential prognostic and therapeutic target for lung adenocarcinoma. World journal of surgical oncology 2018 — PMC5816374.
- Immunological and Functional Characterization of RhoGDI3 and Its Molecular Targets RhoG and RhoB in Human Pancreatic Cancerous and Normal Cells. PloS one 2016 — PMC5104321.
- PubMed PMID:1620121 — UniProt-cited evidence.
- PubMed PMID:8464478 — UniProt-cited evidence.
- PubMed PMID:8810274 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RHOG (Rho-related GTP-binding protein RhoG). Expected localisation Cytoplasmic staining in immune cells (HPA tissue IHC); membrane localization at the molecular level (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A03485 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic 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 A03485); 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-RHOG, 5 μg/mL (datasheet A03485); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining varies across immune-cell subsets (HPA tissue IHC); Isoform / epitope: No isoforms; mature chain spans residues 1–188 (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt P84095, HPA tissue IHC · Enhanced, HPA cell-line RNA · Highest expression in HEL: 215.5 nTPM, Datasheet A03485, PMC4232683. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RHOG/immunohistochemistry-ihc-guide.