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RGS7 / Regulator of G protein signaling 7 · IHC design guide
Design Immunohistochemistry for RGS7
Plan chromogenic RGS7 IHC around high cerebellar GLUC-cell staining and medium neuronal staining in caudate and cortex (HPA tissue IHC). This guide covers FFPE handling, brain-region controls and interpretation of the cytoplasmic/membrane pattern (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RGS7 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 | Cytoplasm/membrane in cerebellar GLUC cells (HPA tissue IHC) | |
| Staining pattern | High in cerebellar GLUC cells; medium in caudate and cortical neurons (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A04305) | |
| Positive control | Cerebellum+2 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
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 A04305) | |
| Caveat | Low hippocampal staining may complicate interpretation (HPA tissue IHC) | |
| Regulation | Brain-specific expression (HPA tissue IHC) | |
| Isoform / epitope | Five isoforms; check antibody epitope coverage (UniProt) |
Section 1
Recommended RGS7 IHC & IF Protocols
The catalog antibody’s paraffin IHC protocol is complemented by three published chromogenic RGS7 IHC protocols using tissue microarrays or brain sections (PMC5766496; PMC5127842; PMC4060866).
| Sample | Paraffin-embedded human Gastric adenocarcinoma tissue; fixative not specified (datasheet A04305) |
| Fixation | Image fixative and duration unreported (datasheet A04305); 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 A04305); 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-RGS7, 1:50 recommended; image 1:200 (datasheet A04305) |
| 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 | RGS7-positive staining in gLUC cells - cytoplasm/membrane of cerebellum (HPA tissue IHC: High). HPA tissue profile: Brain-specific expression. No signal in the no-primary control. |
💡Decision noteStart with heat retrieval in Tris-EDTA pH 9.0 for the catalog antibody (datasheet A04305); a published tissue microarray protocol also uses high pH retrieval (PMC5766496).
Section 2
What Is the Expected RGS7 Staining Pattern?
RGS7 should stain mainly neuronal cytoplasm, with membrane staining possible: HPA reports High cytoplasmic/membranous staining in cerebellar GLUC cells and Medium staining in caudate and cerebral cortex neuronal cells (HPA tissue IHC). HPA rates the tissue pattern Enhanced for consistency with RNA expression (HPA tissue IHC). RGS7 has no transmembrane segment, but interacting proteins can promote membrane localization (UniProt P49802 topology and subunit).
| Cerebellar GLUC cells show strong cytoplasmic staining, with some membrane accentuation; neuronal cells stain in caudate or cortex. | This fits the reported tissue pattern: cerebellar GLUC cells are High in cytoplasm/membrane, while caudate and cortical neuronal cells are Medium (HPA tissue IHC). Membrane staining is plausible because PKD1 or RGS7BP interaction promotes RGS7 membrane localization (UniProt P49802 subcellular location). |
| Staining is predominantly nuclear, with little cytoplasmic or membrane signal in expected positive cells. | A predominantly nuclear pattern conflicts with UniProt's cytoplasmic and membrane locations and HPA's cerebellar cytoplasm/membrane observation (UniProt P49802 subcellular location; HPA tissue IHC). Review the control pattern and antibody-dependent signal before calling nuclei RGS7-positive (general IHC practice). |
| Strong chromogen appears in adipocytes, glandular cells, or respiratory epithelium. | These cells were Not detected in the corresponding HPA adipose, adrenal or appendix, and bronchus tissue examples (HPA tissue IHC). Treat staining as unexpected; antibody cross-reactivity or endogenous detection activity are possibilities to investigate, not established explanations (general IHC practice). |
| Color spreads across multiple compartments or tissue regions without a clear cell-associated pattern. | Diffuse background cannot establish RGS7 localization or cell-specific expression (general IHC practice). Compare a no-primary control with the expected neuronal pattern; nonspecific antibody binding, incomplete blocking, or detection background may contribute (general IHC practice). |
| No convincing staining appears in a cerebellar section expected to contain GLUC cells. | HPA reports High staining in cerebellar GLUC cells, so this is a failed or discrepant positive-control result rather than evidence that RGS7 is universally absent (HPA tissue IHC). Check that the relevant cells are present, then review the IHC detection workflow (general IHC practice). |
💡Expected RGS7 appearanceCall a section positive when cerebellar GLUC cells show High cytoplasmic/membranous staining, or caudate/cortical neuronal cells show Medium staining; dominant nuclear or strong staining in HPA Not detected cell populations is suspect (HPA tissue IHC; UniProt P49802 subcellular location).
| Tissue and cell selection | Cerebellar GLUC cells provide the strongest listed positive example; caudate and cortical neuronal cells are Medium, while hippocampal neuronal cells are Low (HPA tissue IHC). Compare the same cell types when judging intensity, because these reported levels differ (HPA tissue IHC). |
| Intracellular distribution | UniProt lists cytosol/cytoplasm and membrane, and reports that PKD1 or RGS7BP interaction promotes membrane location (UniProt P49802 subcellular location). RGS7 has no transmembrane segment, so membrane accentuation does not imply a membrane-spanning protein (UniProt P49802 topology). |
| Antibody validation | CAB017561 is listed as IHC Enhanced, and HPA describes the tissue result as highly consistent with RNA expression (HPA antibodies; HPA tissue IHC). These assessments support the reported pattern; evaluate unexpected staining with controls rather than assuming every colored structure identifies RGS7 (general IHC practice). |
| Isoforms and processing | UniProt lists 5 isoforms, no signal peptide or propeptide, and a chain spanning residues 1–495 (UniProt P49802). The supplied sources do not map the antibody epitope or establish which isoforms its IHC staining detects (HPA antibodies; UniProt P49802). |
| IF/ICC Q: What localization should I expect? | A: HPA reports mainly supported cytosolic localization in ICC-IF, with mitochondrial localization marked uncertain; its ICC antibody status is Supported (HPA subcellular ICC-IF; HPA antibodies). Interpret the mitochondrial observation cautiously and do not transfer its certainty to paraffin-section IHC (HPA subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| Cerebellar positive control has no detectable chromogen. | The result conflicts with HPA's High GLUC-cell observation; cell sampling or an IHC workflow failure is possible (HPA tissue IHC; general IHC practice). | Confirm GLUC cells are represented, then review primary-antibody application, detection reagents and chromogen development using routine IHC controls (general IHC practice). |
| Only nuclei stain in a neuronal-rich field. | Predominant nuclear localization conflicts with the supplied RGS7 locations and the reported cerebellar cell pattern (UniProt P49802 subcellular location; HPA tissue IHC). | Compare the cerebellar positive control and a no-primary control; reassess whether the nuclear signal follows the primary antibody or the detection system (general IHC practice). |
| Adipose or bronchial cells stain strongly. | HPA reports adipocytes in adipose tissue and respiratory epithelial cells in bronchus as Not detected (HPA tissue IHC). Cross-reactivity or endogenous detection activity is possible (general IHC practice). | Run a no-primary control and review the detection block; score those cells separately from expected neuronal staining (general IHC practice). |
| Background obscures cell borders and cytoplasm. | Nonspecific binding or background from the chromogenic detection workflow may obscure localization (general IHC practice). | Check the no-primary control, blocking and washing steps; interpret a positive result only where the cell-associated pattern remains clear (general IHC practice). |
| Cortex looks weaker than cerebellum. | That contrast can match the reported levels: cortical neuronal cells are Medium and cerebellar GLUC cells are High (HPA tissue IHC). | Compare like cell types and confirm the cortical neuronal signal is cell-associated before changing the assay based on intensity alone (HPA tissue IHC; general IHC practice). |
| Punctate signal is called mitochondrial RGS7 in a tissue section. | HPA marks mitochondrial localization uncertain in ICC-IF; it does not establish that interpretation for tissue IHC (HPA subcellular ICC-IF). | Describe the observed IHC pattern without assigning an organelle; assess any mitochondrial claim separately with an appropriate localization control (general IHC practice). |
Experiment evidence
IHC & IF images showing RGS7 staining
A04305 · IHCImmunohistochemical analysis of paraffin-embedded human Gastric adenocarcinoma. 1, Antibody was diluted at 1:200(4° overnight). 2, Tris-EDTA
A04305-1 · IFIF analysis of RGS7 using anti-RGS7 antibody (A04305-1). RGS7 was detected in an immunocytochemical section of SiHa cells. Enzyme antigen re
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000182901-RGS7/tissue/cerebral+cortex#imid_19944501
HPA · HippocampusHuman Protein Atlas IHC, Hippocampus. Source: https://www.proteinatlas.org/ENSG00000182901-RGS7/tissue/hippocampal+formation#imid_19944486
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000182901-RGS7/tissue/colon#imid_19944411
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000182901-RGS7/tissue/liver#imid_19944428
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody CAB017561; green = RGS7, blue = nucleus, red = microtubules.
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody CAB017561; green = RGS7, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RGS7 IHC & IF
🧪Run cerebellum first; GLUC cells should show cytoplasmic and membrane staining (HPA: High in cerebellar GLUC cells). Use adipose tissue as a negative comparison, focusing on adipocytes (HPA: Not detected in adipocytes). On the cerebellum slide, cells without GLUC staining should remain at background level; the supplied HPA row does not identify a specific internal negative cell type.
Positive control tissue: Cerebellum (GLUC cells - cytoplasm/membrane, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RGS7 in Rh30, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and isotype controls, with the isotype matched to the primary antibody’s host species, immunoglobulin class and clonality; use RGS7 knockout tissue or a validated peptide-blocking control if available (standard IHC practice). For chromogenic detection, quench endogenous peroxidase and inspect the cerebellum control slide for background staining (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the fixative is unreported in the selected A04305 paraffin-section caption (A04305 tissue-IHC caption). That caption uses Tris-EDTA at pH 9.0 for retrieval in gastric adenocarcinoma; retrieval conditions for cerebellum need optimization (A04305 tissue-IHC caption). The supplied evidence does not establish whether frozen sections or IF are easier; neural tissue autofluorescence can complicate IF interpretation (standard IF practice).
HPA tissue IHC evidence for RGS7
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 |
|---|---|---|---|---|
| Cerebellum | GLUC cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RGS7 IHC Tips
Troubleshoot RGS7 staining in paraffin sections by checking retrieval, compartment, cell type and controls before comparing signal intensity.
What retrieval should I use if RGS7 staining is weak in paraffin sections?
Start with heat-mediated Tris-EDTA retrieval at pH 9.0 (datasheet A04305). The selected paraffin-section image used that buffer, a 1:200 primary antibody dilution overnight at 4°C, and a 1:200 secondary antibody for 45 minutes at room temperature (caption A04305). If staining is weak, check heating consistency and section attachment before testing a different retrieval buffer on adjacent sections (standard IHC practice). Compare a candidate condition against the stated method using the same detection settings and a neuronal region with reported RGS7 staining, such as cerebellum (caption A04305; HPA tissue IHC).
Could fixation explain inconsistent RGS7 staining across paraffin blocks?
The selected paraffin-section caption does not state a fixative, and target-specific fixation sensitivity is unknown (caption A04305). Record each block’s fixative and fixation time, then compare matched sections with identical pH 9.0 retrieval and antibody conditions (datasheet A04305; standard IHC practice). Excessive fixation or variable processing can change antigen accessibility in paraffin sections, so assess morphology alongside signal rather than treating an intensity shift alone as altered RGS7 expression (standard IHC practice). Use a positive-control section in each run; the HPA reports high staining in cerebellar GLUC-cell cytoplasm and membrane (HPA tissue IHC).
Which staining compartments are plausible for RGS7 in tissue sections?
Evaluate cytoplasmic and membrane-associated staining because RGS7 is annotated in the cytosol and at membranes, with no transmembrane segment (UniProt P49802 localisation and topology). Interactions with PKD1 or RGS7BP can promote membrane localisation, so a membrane rim is plausible but should be judged against cellular morphology (UniProt P49802 interactions; standard IHC practice). HPA reports high cytoplasmic/membrane staining in cerebellar GLUC cells and mainly cytosolic subcellular localisation; its additional mitochondrial localisation is marked uncertain (HPA tissue IHC; HPA subcellular). If the signal appears exclusively nuclear or follows tissue edges, review controls and section quality before assigning it to RGS7 (UniProt P49802 localisation; standard IHC practice).
Can one RGS7 antibody report every isoform and phosphorylation state?
RGS7 has 5 annotated isoforms, but the supplied record does not map the catalog antibody’s epitope onto their sequences (UniProt P49802 isoforms; caption A04305). Ask for epitope coordinates and compare them with the intended isoforms before describing staining as total RGS7 (standard IHC practice). The protein has annotated phosphosites at residues 229, 241, 243 and 434; their presence alone does not establish an effect on antibody binding (UniProt P49802 modified residues). If two antibodies give different patterns, compare their mapped epitopes and stain adjacent sections under matched retrieval and detection conditions (standard IHC practice).
How should I cross-check chromogenic RGS7 staining with multiplex IF?
Use IF as a separate validation experiment, pairing RGS7 with a marker that identifies the expected neuronal cell population in the sampled region (HPA tissue IHC; standard IF practice). HPA reports neuronal staining in caudate and cerebral cortex, while cerebellar GLUC cells show high cytoplasmic/membrane staining (HPA tissue IHC). Select a fluorophore channel with low measured tissue autofluorescence and include single-stain and secondary-only controls before interpreting overlap (standard IF practice). Because RGS7 has no transmembrane segment and is mainly cytosolic, permeabilise sufficiently for an intracellular epitope; if the antibody epitope is unknown, establish its accessibility empirically (UniProt P49802 topology; HPA subcellular; standard IF practice).
How can I reduce diffuse or punctate background in RGS7 IHC?
Check a no-primary control for detection-system staining, and inspect the section for pigment, folds and precipitate before changing the RGS7 antibody concentration (standard IHC practice). For peroxidase-based chromogenic detection, apply an endogenous peroxidase block and compare the resulting background with the no-primary section (standard IHC practice). Titrate the primary around the caption’s 1:200 condition while keeping pH 9.0 retrieval and detection settings constant (caption A04305; datasheet A04305; standard IHC practice). HPA reports RGS7 as not detected in adipocytes of adipose tissue, providing a tissue-pattern comparison rather than proof that every stained cell is background (HPA tissue IHC).
What should I score when comparing RGS7 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; RGS7 can appear in the cytoplasm or at membranes (UniProt P49802 localisation; standard IHC practice). For a defined population, report the percentage of positive cells and an intensity-based H-score, using the same positivity threshold across slides (standard IHC practice). If cell counts vary greatly, also report positive-cell density per mm² of viable, assessable tissue and exclude folds or necrosis from the measured area (standard IHC practice). Normalise counts to the number of eligible cells, or density to the measured tissue area, and run a shared control section to monitor batch variation (standard IHC practice).
How do I distinguish credible RGS7 staining from artefact?
A credible pattern should fit the sampled cells and cytoplasmic or membrane distribution; HPA reports high staining in cerebellar GLUC cells and medium neuronal staining in caudate and cerebral cortex (HPA tissue IHC; UniProt P49802 localisation). Treat isolated nuclear staining, section-edge enhancement, necrotic-area staining and signal reproduced in a no-primary control as reasons to investigate artefact (UniProt P49802 localisation; standard IHC practice). Check endogenous peroxidase activity and pigment when a chromogenic deposit appears in unexpected cells (standard IHC practice). The selected image shows a paraffin-embedded gastric adenocarcinoma section, but its caption alone does not establish cell-specific RGS7 expression there (caption A04305).
Boster reagents
Best RGS7 / Regulator of G protein signaling 7 IHC Antibodies
Anti-RGS7 antibodies have IHC data from human paraffin-embedded gastric adenocarcinoma (A04305 image caption) and IF/ICC data from SiHa cells (A04305-1 image caption); both list Human, Mouse, and Rat reactivity (catalog).
Real IHC data
Anti-RGS7 Antibody
Cat # A04305
Real IF data
Anti-RGS7 Antibody ®
Cat # A04305-1
A04305 will render with its IHC image of paraffin-embedded human gastric adenocarcinoma (A04305 image caption); its listed applications include IHC (catalog). A04305-1 will render with its IF image of SiHa cells (A04305-1 image caption); its listed applications include IF and ICC (catalog).
Which to pick: For paraffin-section IHC, choose A04305: it lists IHC and is polyclonal (catalog), and its image caption shows paraffin-embedded human gastric adenocarcinoma but does not report the fixative (A04305 image caption). For IF/ICC, choose A04305-1: it lists both applications (catalog) and has a SiHa cell IF image (A04305-1 image caption). For cross-species planning, both list Human, Mouse, and Rat reactivity (catalog); the A04305-1 IF/ICC dilution entry specifies Human (catalog dilution entry).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P49802 (RGS7_HUMAN, Regulator of G protein signaling 7).
- Human Protein Atlas. RGS7 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. RGS7 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the mitochondria..
- Human Protein Atlas. RGS7 antibody validation summary (1 antibodies).
- RGS7 is recurrently mutated in melanoma and promotes migration and invasion of human cancer cells. Scientific reports 2018 — PMC5766496.
- Cellular and Subcellular Localization of the RGS7/Gβ5/R7BP Complex in the Cerebellar Cortex. Frontiers in neuroanatomy 2016 — PMC5127842.
- Association of Rgs7/Gβ5 complexes with Girk channels and GABAB receptors in hippocampal CA1 pyramidal neurons. Hippocampus 2013 — PMC4060866.
- Sensitivity and kinetics of signal transmission at the first visual synapse differentially impact visually-guided behavior. eLife 2015 — PMC4412108.
- PubMed PMID:10339594 — UniProt-cited evidence.
- PubMed PMID:8548815 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RGS7 (Regulator of G protein signaling 7). Expected localisation Cytoplasm/membrane in cerebellar GLUC cells (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A04305). Validated IHC antibody: A04305 at 1:50 recommended; image 1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (GLUC cells - cytoplasm/membrane); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human Gastric adenocarcinoma tissue; fixative not specified (datasheet A04305); 2. Fixation: Image fixative and duration unreported (datasheet A04305); 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 A04305); 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-RGS7, 1:50 recommended; image 1:200 (datasheet A04305); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04305); Caveat: Low hippocampal staining may complicate interpretation (HPA tissue IHC); Isoform / epitope: Five isoforms; check antibody epitope coverage (UniProt); Regulation: Brain-specific expression (HPA tissue IHC). Evidence: UniProt P49802, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A04305, PMC5766496, PMC5127842. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RGS7/immunohistochemistry-ihc-guide.