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RGS10 / Regulator of G protein signaling 10 · IHC design guide
Design Immunohistochemistry for RGS10
Plan chromogenic IHC for RGS10 using the cytoplasmic and nuclear tissue pattern (HPA tissue IHC). This guide identifies lymphoid tissues as high-staining references (HPA tissue IHC) and covers fixation, retrieval, and staining controls.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RGS10 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 and nuclear staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic and nuclear; most abundant in lymphoid tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A05197-1) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Epididymis+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Forskolin promotes nuclear translocation (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended RGS10 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A05197-1) and one published RGS10 paraffin-section protocol (PMC11326775) provide starting conditions.
| Sample | Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A05197-1) |
| Fixation | Image fixative and duration unreported (datasheet A05197-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A05197-1); 20 min, 95–100 °C (standard) |
| 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-RGS10, 1:100-1:300 (datasheet A05197-1) |
| 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 | RGS10-positive staining in lymphoid tissue of appendix (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic and nuclear expression, most abundant in lymphoid tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA retrieval at pH 9.0 (datasheet: A05197-1). The published method does not specify retrieval (PMC11326775: methods).
Section 2
What Is the Expected RGS10 Staining Pattern?
RGS10 can stain the cytoplasm and nucleus, consistent with a protein lacking a transmembrane segment (UniProt O43665: localization and topology). In paraffin sections, expect the strongest representative staining in lymphoid tissue, including lymph node germinal center cells, alongside other HPA high populations (HPA: tissue IHC). HPA rates its tissue IHC evidence Enhanced, while noting only medium consistency between antibody staining and RNA expression (HPA: reliability description).
| Cytoplasmic and nuclear stain appears in lymph node germinal center cells or appendix lymphoid tissue. | This fits the reported compartments and high staining in these populations (UniProt O43665: localization; HPA: tissue IHC). Score the named cell population and its compartment rather than calling the entire section uniformly positive (standard IHC practice). |
| Stain appears only on cell borders or in extracellular material, without a convincing intracellular pattern. | Recheck localization: UniProt lists cytosol and nucleus and no transmembrane segment (UniProt O43665: localization and topology). Border staining alone does not establish RGS10; compare morphology, controls and the specific population before accepting it (standard IHC practice). |
| Strong stain dominates a population listed as not detected, such as skeletal muscle myocytes. | That conflicts with the reported IHC observation for that population (HPA: not detected in skeletal muscle myocytes). Consider antibody cross-reactivity or endogenous detection activity, then inspect a reagent control (standard IHC practice). HPA not detected is not proof that expression is impossible. |
| Color spreads diffusely across cells and surrounding tissue, obscuring nuclear and cytoplasmic boundaries. | The distribution cannot be scored reliably against the reported intracellular pattern (HPA: tissue IHC; UniProt O43665: localization). Review reagent controls, blocking and detection background before interpreting intensity (standard IHC practice). |
| No convincing stain appears in lymph node germinal center cells or bone marrow hematopoietic cells. | Both populations are reported High, so an absent result calls for an assay check (HPA: tissue IHC). Confirm that the chosen population is present, then review the antibody, retrieval and detection controls (standard IHC practice); tissue positivity is not guaranteed in every specimen. |
💡Expected RGS10 appearanceA credible positive shows discernible cytoplasmic and/or nuclear stain in named HPA High cells, especially lymphoid populations; isolated extracellular color or dominant stain in a reported not-detected population needs control review (HPA: tissue IHC; UniProt O43665: localization; standard IHC practice).
| Tissue and cell selection | Appendix lymphoid tissue and lymph node germinal center cells are High; several glandular populations are Low or not detected (HPA: tissue IHC). Identify the cell population before comparing signal intensity across sections (standard IHC practice). |
| Localization and stimulation | UniProt lists cytosol and nucleus and says forskolin promotes phosphorylation and nuclear translocation (UniProt O43665: localization). Nuclear predominance may depend on treatment; it should not be required of untreated tissue. |
| Antibody validation | HPA021305 and CAB004559 each have Enhanced IHC status (HPA: antibody validation). HPA describes the tissue profile as having medium antibody–RNA consistency (HPA: reliability description); neither label establishes specificity in every specimen. |
| Isoform interpretation | UniProt lists three RGS10 isoforms (UniProt O43665: isoforms). The supplied evidence gives no epitope mapping, so staining alone cannot distinguish isoforms or establish that every isoform is detected. |
| IF/ICC Q: What localization is reported? | A: HPA reports supported nucleoplasmic localization and additional nuclear bodies in ICC-IF (HPA: subcellular). This cell-image observation can inform interpretation; it is not an IHC protocol or a requirement that every paraffin section show nuclear bodies. |
| Detection background | Endogenous enzyme activity or nonspecific reagent binding can produce chromogenic signal (standard IHC practice). A reagent control helps separate that background from cell-associated staining; the supplied sources do not quantify either effect for RGS10. |
| Situation | Likely cause | Next action |
|---|---|---|
| The high-expression reference tissue looks blank. | The expected population may be absent from the field, or the IHC assay may have failed (HPA: High in lymph node germinal center and bone marrow hematopoietic cells; standard IHC practice). | Locate the named cells on a counterstained section, then check antibody, retrieval and detection steps with appropriate controls (standard IHC practice). Do not infer biological absence from one blank field. |
| A reported not-detected population stains strongly. | Cross-reactivity or endogenous chromogenic activity is possible (standard IHC practice); HPA reports no detection in skeletal muscle myocytes and ovarian stroma cells (HPA: tissue IHC). | Check tissue identity and morphology, compare a reagent control, and assess staining with an independently validated antibody if available (standard IHC practice). Treat the HPA category as a comparison, not an absolute exclusion. |
| Color is diffuse and compartment boundaries cannot be scored. | Background from blocking, washing or detection may obscure the intracellular pattern (standard IHC practice; UniProt O43665: localization). | Inspect reagent controls and background outside cells; adjust blocking, washing or detection conditions using standard IHC practice before assigning an RGS10 score. |
| Stain is confined to an apparently extracellular deposit. | That pattern does not match the cytosolic and nuclear localization or lack of a transmembrane segment (UniProt O43665: localization and topology). | Check morphology and reagent controls, then seek reproducible intracellular staining in an HPA High cell population before calling the deposit RGS10 (HPA: tissue IHC; standard IHC practice). |
| A nucleus-positive field differs from a mainly cytoplasmic field. | Both compartments are reported, and forskolin can promote nuclear translocation (UniProt O43665: localization). Differences alone cannot establish treatment history or a technical fault. | Record nuclear and cytoplasmic scores separately and compare like cell populations and known treatment conditions (standard IHC practice). Avoid inferring stimulation solely from stain location. |
| Nuclear bodies are absent in paraffin IHC. | HPA's supported nuclear-body observation comes from ICC-IF images, while tissue IHC is summarized as general cytoplasmic and nuclear expression (HPA: subcellular; HPA: tissue IHC). | Judge the paraffin result against its tissue IHC pattern and controls (HPA: tissue IHC; standard IHC practice). Do not require ICC-IF nuclear bodies to validate chromogenic IHC. |
Experiment evidence
IHC & IF images showing RGS10 staining
A05197-1 · IHCImmunohistochemical analysis of paraffin-embedded human tonsil. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA,pH9.0 was used
A05197 · IHCImmunohistochemistry (IHC) analyzes of RGS10 pAb in paraffin-embedded human tonsil carcinoma tissue at 1:50.showing cytoplasmic and nucleus
A05197 · IHCImmunohistochemistry (IHC) analyzes of RGS10 pAb in paraffin-embedded human stomach carcinoma tissue at 1:50.showing cytoplasmic and nucleus
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000148908-RGS10/tissue/cerebral+cortex#imid_2246640
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000148908-RGS10/tissue/colon#imid_2246566
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000148908-RGS10/tissue/liver#imid_2246583
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000148908-RGS10/tissue/kidney#imid_2246568
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA021305; green = RGS10, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA021305; green = RGS10, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RGS10 IHC & IF
🧪Run appendix first and score staining in its lymphoid tissue (HPA: High in appendix lymphoid tissue). Run epididymis as the negative tissue and assess its glandular cells (HPA: Not detected in epididymis glandular cells); on the appendix slide, use morphologically distinct unstained cells to gauge background without assuming they are RGS10-negative.
Positive control tissue: Appendix (Lymphoid tissue, HPA High)
Negative control tissue: Epididymis (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RGS10 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host and immunoglobulin class; use a confirmed RGS10 knockout sample as a biological negative if available (standard IHC practice). Block endogenous peroxidase and check background in the appendix section before interpreting chromogenic staining (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected SKU A05197-1 tissue-IHC caption does not state its fixative (selected-SKU caption: fixative unreported). That caption uses pH 9.0 Tris-EDTA retrieval on a paraffin section, but provides no comparison establishing retrieval dependence (selected-SKU caption: Tris-EDTA, pH 9.0). ICC-IF images exist for A-431, U-251MG and U2OS (HPA: ICC-IF image cell lines); the supplied evidence does not establish that IF or frozen sections are easier for this target.
HPA tissue IHC evidence for RGS10
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 |
|---|---|---|---|---|
| Appendix | Lymphoid tissue | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | Goblet cells | High | Protein (IHC) | HPA → |
| Cerebellum | Granular cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Prostate | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RGS10 IHC Tips
Troubleshoot RGS10 staining in paraffin-section chromogenic IHC using the catalog antibody’s tonsil image, reported tissue patterns, and appropriate section controls.
Which retrieval condition should I start with for RGS10 in paraffin sections?
Start with heat-mediated Tris-EDTA at pH 9.0 for RGS10 paraffin-section IHC (datasheet A05197-1). The demonstrated human tonsil preparation used antibody at 1:200 overnight at 4°C, so keep antibody exposure constant while comparing retrieval times on adjacent sections (caption A05197-1; standard IHC practice). If staining remains weak, adjust heating time and assess tissue morphology before testing another buffer as a fallback (standard IHC practice). Select the condition that preserves interpretable nuclear and cytoplasmic staining while primary-omission controls stay clean (HPA: general cytoplasmic and nuclear expression; standard IHC practice).
Could fixation explain weak or uneven RGS10 staining?
Target-specific fixation sensitivity is unknown because the selected paraffin-embedded tonsil caption does not state its fixative (caption A05197-1). Record each sample’s fixative and fixation duration, then compare 2 similarly processed adjacent sections under the same retrieval and detection conditions (standard IHC practice). If staining differs, assess section quality and processing history before assigning the difference to RGS10 antigen loss (standard IHC practice). Keep the antibody at the demonstrated 1:200 condition for the initial comparison, and include a consistently processed positive-control section so changes in tissue handling remain distinguishable from assay drift (caption A05197-1; standard IHC practice).
Should RGS10 staining be nuclear, cytoplasmic, or both?
Score nuclear and cytoplasmic staining separately because RGS10 is reported in cytosol and nucleus, while tissue IHC shows general cytoplasmic and nuclear expression (UniProt O43665 localisation; HPA: tissue profile). A nuclear signal is plausible, but its presence alone does not demonstrate pathway activation or a specific treatment response (UniProt O43665: forskolin-associated nuclear translocation; standard IHC interpretation). Compare the 2 compartments within the same identified cell population and retain a hematoxylin counterstain to define nuclei (standard IHC practice). Treat exclusively rim-like staining cautiously because the record reports no transmembrane segment, and check it against primary-omission and morphology controls (UniProt O43665 topology; standard IHC practice).
How can isoforms or epitope position affect RGS10 IHC interpretation?
RGS10 has 3 annotated isoforms, but the supplied antibody caption does not identify its epitope or establish which isoforms it recognizes (UniProt O43665 isoforms; caption A05197-1). Map the antibody immunogen to the isoform sequences before calling staining isoform-specific, and check whether it overlaps the RGS domain at residues 41–156 (UniProt O43665 domains; standard antibody-validation practice). Reported phosphoserines at 24, 41, and 176 provide positions to examine during epitope review, not evidence that phosphorylation alters this antibody’s binding (UniProt O43665 modified residues). Resolve uncertain patterns with an independent epitope antibody or a validated loss-of-target control under matched IHC processing (standard IHC practice).
How should companion IF help interpret a chromogenic RGS10 result?
Use companion IF as a localization check, scoring RGS10 alongside a validated marker for the expected cell population, such as the non-germinal-center population highlighted in tonsil IHC (HPA: high in tonsil non-germinal-center cells; standard IF practice). Choose 2 well-separated fluorophore channels after checking unstained tissue for autofluorescence, and include single-color controls to assess spectral bleed-through (standard IF practice). Because RGS10 is intracellular and has no transmembrane segment, permeabilize sufficiently to reach cytosolic and nuclear epitopes while preserving morphology (UniProt O43665 localisation and topology; standard IF practice). Compare compartment patterns with the chromogenic section, but establish IF fixation and antibody conditions independently because the selected IHC caption reports neither an IF protocol nor a fixative (caption A05197-1; standard IF practice).
What should I check when RGS10 DAB staining appears widespread?
First compare the stained section with primary-omission and detection-only controls to identify nonspecific reagent deposition or residual endogenous peroxidase activity (standard chromogenic IHC practice). A peroxidase block and controlled DAB development are general workflow steps; neither is evidence of RGS10 expression (standard chromogenic IHC practice). Recheck antibody concentration against the documented 1:200 tonsil condition, then optimize blocking, washing, and detection time one variable at a time (caption A05197-1; standard IHC practice). Interpret diffuse staining against cell identity and morphology, since tissue IHC reports abundant lymphoid expression but no detection in skeletal-muscle myocytes (HPA: tissue profile; HPA: skeletal muscle myocytes not detected).
How should I quantify RGS10 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region of interest before scoring, then record nuclear and cytoplasmic RGS10 separately because both compartments are reported (HPA: general cytoplasmic and nuclear expression; standard IHC practice). For an H-score, multiply the percentage of cells at each intensity from 0–3 by that intensity and sum to a 0–300 score (standard IHC scoring practice). Alternatively report percent positive cells among eligible cells or positive-cell density per mm² of evaluable tissue, stating the threshold used (standard IHC scoring practice). Normalize comparisons to the same cell type, evaluable area, staining run, and positive-control performance, especially when tissue composition differs (standard IHC practice).
How can I distinguish true RGS10 staining from artefact?
Expect a cell-associated nuclear or cytoplasmic pattern, and check whether the stained population matches the annotated tissue pattern rather than assuming every brown deposit is RGS10 (UniProt O43665 localisation; HPA: tissue profile; standard IHC interpretation). In tonsil, non-germinal-center cells are reported high, while skeletal-muscle myocytes are reported not detected, providing context for positive and low-expression comparisons (HPA: tonsil; HPA: skeletal muscle). Inspect 2 control types—an appropriate positive tissue and a primary-omission section—alongside the sample (standard IHC practice). Edge staining, necrotic areas, pigment, and residual endogenous-enzyme signal require morphological and control review before inclusion in a score (standard chromogenic IHC practice).
Boster reagents
Best RGS10 / Regulator of G protein signaling 10 IHC Antibodies
Two anti-RGS10 antibodies have IHC figures from human paraffin sections (catalog IHC image captions); one also lists IF and human, mouse, and rat reactivity (A05197-1 applications/reactivity).
Real IHC data
Anti-RGS10 Antibody
Cat # A05197-1
Real IHC data
Anti-RGS10 Antibody
Cat # A05197
A05197-1 shows IHC in paraffin-embedded human tonsil (A05197-1 IHC image caption) and lists IF plus human, mouse, and rat reactivity (A05197-1 applications/reactivity). A05197 shows IHC in paraffin-embedded human tonsil carcinoma and stomach carcinoma (A05197 IHC image captions) and lists human reactivity (A05197 reactivity).
Which to pick: For tissue IHC, choose A05197-1 for its human tonsil example or A05197 for its human carcinoma examples (catalog IHC image captions); both captions show paraffin sections, but neither reports the fixative (catalog IHC image captions). For IF/ICC planning, choose polyclonal A05197-1 because it lists IF; ICC validation and an IF figure are unreported (A05197-1 dilution_raw; applications; if_image_alts). For mouse or rat samples, choose A05197-1 based on its listed reactivity (A05197-1 reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O43665 (RGS10_HUMAN, Regulator of G protein signaling 10).
- Human Protein Atlas. RGS10 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. RGS10 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nuclear bodies..
- Human Protein Atlas. RGS10 antibody validation summary (2 antibodies).
- RGS10 deficiency facilitates distant metastasis by inducing epithelial-mesenchymal transition in breast cancer. eLife 2024 — PMC11326775.
- Short stature and combined immunodeficiency associated with mutations in RGS10. Science signaling 2021 — PMC8522579.
- RGS10 deficiency ameliorates the severity of disease in experimental autoimmune encephalomyelitis. Journal of neuroinflammation 2016 — PMC4736282.
- Microglia Exhibit Distinct Heterogeneity Rather than M1/M2 Polarization within the Early Stage of Acute Ischemic Stroke. Aging and disease 2023 — PMC10676790.
- PubMed PMID:8774883 — UniProt-cited evidence.
- PubMed PMID:10791963 — UniProt-cited evidence.
- PubMed PMID:11443111 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RGS10 (Regulator of G protein signaling 10). Expected localisation Cytoplasmic and nuclear staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A05197-1). Validated IHC antibody: A05197-1 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Lymphoid tissue); negative Epididymis. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A05197-1); 2. Fixation: Image fixative and duration unreported (datasheet A05197-1); 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: Tris-EDTA pH 9.0 (datasheet A05197-1); 20 min, 95–100 °C (standard); 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-RGS10, 1:100-1:300 (datasheet A05197-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unspecified (UniProt); Regulation: Forskolin promotes nuclear translocation (UniProt). Evidence: UniProt O43665, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A05197-1, PMC11326775. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RGS10/immunohistochemistry-ihc-guide.