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RPS20 / Small ribosomal subunit protein uS10 · IHC design guide
Design Immunohistochemistry for RPS20
Plan RPS20 chromogenic IHC on paraffin sections using the reported general cytoplasmic tissue pattern (HPA tissue IHC). Compare high-staining glandular or neuronal cells with skeletal myocytes, where staining was not detected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RPS20 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 | General cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in glandular and neuronal cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Skeletal muscle |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Skeletal myocytes are not detected (HPA tissue IHC) | |
| Regulation | Specific expression regulators unannotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope effect undetermined (UniProt) |
Section 1
Recommended RPS20 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet) is accompanied by published RPS20 IHC protocols for renal carcinoma and glioblastoma tissue (PMC9773015; PMC4624638).
| Sample | Paraffin-embedded human kidney tissue; fixative not specified (datasheet M06634) |
| Fixation | Image fixative and duration unreported (datasheet M06634); 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 monoclonal (clone FEH-18) anti-RPS20, 1:50 (datasheet M06634) |
| 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 | RPS20-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval rule); adjust using the glioblastoma protocol’s pressure-cooker retrieval if needed (PMC4624638: methods).
Section 2
What Is the Expected RPS20 Staining Pattern?
RPS20 is a cytoplasmic component of the 40S ribosomal subunit and has no transmembrane segment (UniProt P60866). Expect cytoplasmic staining in many cell types, including appendix glandular cells, bone marrow hematopoietic cells and cortical neurons, where HPA reports High staining (HPA tissue IHC). HPA describes the overall pattern as general cytoplasmic expression and rates its IHC staining Approved, supported by external characterization data (HPA tissue IHC).
| Cytoplasmic staining appears in appendix glandular cells or bone marrow hematopoietic cells. | This matches the expected compartment (UniProt P60866; HPA tissue IHC). HPA reports High staining in both cell populations, making them useful positive references for slide review (HPA tissue IHC). |
| Signal is mainly nuclear or outlines the plasma membrane, with little cytoplasmic staining. | That distribution conflicts with UniProt's cytoplasmic assignment and HPA's general cytoplasmic IHC profile (UniProt P60866; HPA tissue IHC). Treat it as a possible staining artefact and assess controls before calling it RPS20. |
| Skeletal muscle myocytes stain strongly while expected positive cells stain weakly. | HPA reports RPS20 as Not detected in skeletal muscle myocytes (HPA tissue IHC). Unexpected signal may reflect cross-reactivity or endogenous chromogenic activity (general IHC practice); check controls before assigning target expression. |
| Brown signal spreads across tissue, empty spaces or slide background without clear cell boundaries. | A broad deposit cannot establish the reported cytoplasmic cell pattern (HPA tissue IHC). In chromogenic IHC, nonspecific binding, endogenous detection activity or excess detection reagent can cause background (general IHC practice). |
| No stain appears in appendix glandular cells or bone marrow hematopoietic cells. | Both are reported High by HPA, so an absent signal calls the staining run into question (HPA tissue IHC). Review tissue preservation, antibody application, retrieval and detection controls as general IHC checks; this record does not establish RPS20 fixation sensitivity. |
💡Expected RPS20 appearanceCall the stain positive when cytoplasmic signal is clear in HPA High cells such as appendix glandular cells; isolated nuclear, membrane-rim or acellular deposit is suspect (UniProt P60866; HPA tissue IHC).
| Tissue and cell choice | HPA reports High staining in appendix and breast glandular cells, bone marrow hematopoietic cells and cortical neurons, but Not detected in skeletal muscle myocytes (HPA tissue IHC). Compare named cell populations rather than whole-section darkness. |
| Cytoplasmic target location | RPS20 is a cytoplasmic 40S subunit protein without a transmembrane segment (UniProt P60866). Interpret signal by its cellular compartment; this annotation does not predict a membrane pattern. |
| IHC evidence strength | HPA rates its tissue IHC profile Approved because external characterization supports the staining (HPA tissue IHC). That rating describes the reported antibody evidence; it does not validate every antibody or establish a fixation effect. |
| Isoforms and processing | UniProt lists 2 isoforms and annotates a chain spanning residues 2–119, with no signal peptide or propeptide (UniProt P60866). The payload gives no epitope mapping, so isoform recognition cannot be assigned. |
| IF/ICC cross-check — where should signal appear? | HPA reports supported cytosol and endoplasmic-reticulum localization in ICC-IF images from A-431, U-251MG and U2OS (HPA subcellular ICC-IF). Use this as compartment context; evaluate IHC with the tissue IHC profile. |
| Situation | Likely cause | Next action |
|---|---|---|
| An expected positive tissue is blank. | Appendix glandular cells and bone marrow hematopoietic cells are reported High, so a blank result may indicate an assay failure (HPA tissue IHC). | Confirm the intended cells are present, then review antibody application, retrieval and detection controls (general IHC practice). No RPS20-specific retrieval or fixation sensitivity is supplied. |
| Staining appears mainly in nuclei. | A dominant nuclear pattern differs from the reported cytoplasmic localization (UniProt P60866; HPA tissue IHC). | Check the positive reference and a control lacking primary antibody; reassess localization before scoring (general IHC practice). |
| Skeletal muscle myocytes appear positive. | HPA reports these myocytes as Not detected; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Compare a control lacking primary antibody and confirm signal belongs to myocytes rather than neighboring cells (general IHC practice). |
| Diffuse brown background obscures cells. | Nonspecific antibody binding or endogenous detection activity can obscure the cytoplasmic pattern (general IHC practice; HPA tissue IHC). | Review blocking, washing and detection controls, then adjust routine IHC conditions using the antibody's instructions (general IHC practice). |
| Only some tissue compartments stain. | HPA reports different levels across named cells, including High in cortical neurons and Low in kidney tubule cells (HPA tissue IHC). | Identify and score the relevant cell population against its HPA entry; avoid treating all cells in an organ as one reference (HPA tissue IHC). |
| The slide shows a sharp membrane rim. | RPS20 has no annotated transmembrane segment, and HPA describes general cytoplasmic IHC expression (UniProt P60866; HPA tissue IHC). | Inspect controls and verify that cytoplasmic staining accompanies the rim before calling it target signal (general IHC practice). |
Experiment evidence
IHC & IF images showing RPS20 staining
M06634 · IHCImmunohistochemical analysis of paraffin-embedded human kidney, using RPS20 Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000008988-RPS20/tissue/cerebral+cortex#imid_1260668
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000008988-RPS20/tissue/colon#imid_1260602
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000008988-RPS20/tissue/liver#imid_1260587
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000008988-RPS20/tissue/kidney#imid_1260604
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003570; green = RPS20, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003570; green = RPS20, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RPS20 IHC & IF
🧪Run appendix first: its glandular cells should stain (HPA: Appendix glandular cells, High). Use skeletal muscle myocytes as the negative tissue (HPA: Skeletal muscle myocytes, Not detected); no validated internal negative cell population is identified within the appendix slide (HPA: appendix row identifies glandular cells only).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Skeletal muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RPS20 in A-431, U-251MG, U2OS, with annotated localisation: Endoplasmic reticulum (supported), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, an isotype control matched to the primary antibody’s host species and clonality, and an RPS20 knockout sample or validated peptide-block control (standard IHC practice). Quench endogenous peroxidase in appendix sections and check endogenous biotin if using avidin–biotin detection (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative for the selected paraffin-section kidney image is unreported (M06634 tissue-IHC caption). Retrieval dependency is unreported; optimise antigen retrieval for the catalog antibody on paraffin sections (standard IHC practice). Whether frozen sections or IF/ICC are easier is unreported; appendix inflammatory cells may contribute endogenous peroxidase background (standard IHC practice).
HPA tissue IHC evidence for RPS20
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — External characterization data supports antibody staining.). 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 | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RPS20 IHC Tips
Troubleshoot RPS20 staining in paraffin sections by checking retrieval, compartment, cell identity and controls before comparing signal intensity.
How should I adjust retrieval when RPS20 staining is weak in paraffin sections?
Start with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule). Keep section thickness, cooling time and detection conditions consistent while comparing a weak slide with a previously successful run (standard IHC practice). If staining remains weak, compare a modestly longer retrieval with the starting condition on adjacent sections, watching for tissue damage or rising background (standard IHC practice). Score cytoplasmic signal in the same cell population rather than treating any dark deposit as recovery, because RPS20 is cytoplasmic and part of the 40S subunit (UniProt P60866).
Could fixation explain inconsistent RPS20 staining between paraffin blocks?
Target-specific fixation sensitivity is unknown: the selected kidney image describes paraffin-embedded tissue but does not state its fixative (selected M06634 caption). Compare blocks with documented processing histories, then stain adjacent sections together using the same citrate pH 6.0 retrieval for 20 min (page retrieval rule; standard IHC practice). If a poorly staining block also shows weak control staining or damaged morphology, investigate its processing and detection before attributing the result to RPS20 (standard IHC practice). Record fixation and processing differences in the report; neither the tissue staining profile nor protein annotations establish a fixation effect (HPA tissue IHC; UniProt P60866).
Where should convincing RPS20 chromogenic signal appear?
Expect predominantly cytoplasmic staining, because RPS20 is annotated as cytoplasmic and a component of the 40S ribosomal subunit (UniProt P60866). Subcellular imaging also supports cytosol and endoplasmic reticulum localisation, so a fine cytoplasmic pattern is plausible (HPA subcellular). Compare staining within identifiable cells: HPA reports high signal in appendix glandular cells and low signal in kidney tubule cells (HPA tissue IHC). Predominantly nuclear or sharply membrane-limited deposits warrant a review of morphology, counterstain and detection controls before being assigned to RPS20 (UniProt P60866 topology; standard IHC practice).
How can isoforms or modified residues affect interpretation of this antibody?
RPS20 has 2 annotated isoforms, but the supplied evidence does not locate this antibody’s epitope within either sequence (UniProt P60866; selected M06634 caption). The record lists modifications at residues including 2, 9, 34 and 75; their effect on staining cannot be assigned without epitope mapping (UniProt P60866). If blocks disagree, compare staining after identical retrieval and detection before proposing isoform selection or modification-dependent masking (standard IHC practice). Treat an apparent compartment shift cautiously because the supplied localisation evidence points to cytoplasm, cytosol and endoplasmic reticulum (UniProt P60866; HPA subcellular).
How should an IF follow-up test a puzzling chromogenic RPS20 pattern?
Use IF as a separate follow-up and retain the chromogenic section as the tissue reference; the selected M06634 caption documents paraffin-section IHC only (selected M06634 caption). Multiplex RPS20 with a marker for the expected cell population, chosen for the actual specimen, to separate cell identity from apparent intensity (standard IF practice). Select fluorophores after checking tissue autofluorescence and include single-color controls for bleed-through (standard IF practice). Because RPS20 lacks a transmembrane segment and is associated with cytosol and endoplasmic reticulum, optimise permeabilisation for intracellular access while preserving morphology (UniProt P60866 topology; HPA subcellular; standard IF practice).
What should I check when RPS20 DAB staining looks diffuse or granular?
Compare the suspect section with a no-primary control and inspect whether deposits track tissue folds, cut edges or damaged areas (standard IHC practice). Verify that the peroxidase block worked before interpreting DAB signal, particularly where endogenous enzyme activity could mimic staining (standard chromogenic IHC practice). RPS20’s expected general cytoplasmic distribution can make genuine staining look widespread, so judge individual intact cells and their compartments rather than overall slide darkness (HPA tissue IHC; UniProt P60866). If background persists, adjust blocking, washing or antibody concentration one variable at a time while keeping retrieval at citrate pH 6.0 initially (standard IHC practice; page retrieval rule).
How should I quantify RPS20 staining across regions with different cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, then record the percentage of positive intact cells and intensity using a consistent H-score scheme (standard IHC practice). Normalise counts to the number of eligible cells, or report positive-cell density per mm² when region area is the comparison unit (standard IHC practice). Keep section processing, retrieval and image thresholds consistent across samples, and score folds, edges and necrotic areas separately or exclude them by a predefined rule (standard IHC practice). Report cell type with each score because HPA lists high staining in appendix glandular cells but low staining in kidney tubule cells (HPA tissue IHC).
How can I distinguish true RPS20 staining from a misleading positive?
A credible positive shows staining in intact cells with a predominantly cytoplasmic distribution, consistent with RPS20 annotation and HPA’s general tissue profile (UniProt P60866; HPA tissue IHC). Check cell identity: high appendix glandular-cell staining and low kidney tubule-cell staining are reference patterns, while skeletal-muscle myocytes are reported as undetected (HPA tissue IHC). Treat isolated nuclear or membrane-rim signal, edge enhancement and deposits over necrotic tissue as suspect until morphology and controls support them (UniProt P60866 topology; standard IHC practice). Compare a no-primary control and the peroxidase-block result to identify detection artefacts before concluding that an unexpected pattern reflects RPS20 (standard chromogenic IHC practice).
Boster reagents
Best RPS20 / Small ribosomal subunit protein uS10 IHC Antibodies
The catalog lists M06634 for IHC and IF/ICC in human, mouse, and rat (catalog applications/reactivity). Its image shows IHC on paraffin-embedded human kidney; no IF image is supplied (catalog image captions).
Real IHC data
Anti-RPS20/Ribosomal Protein S20 Rabbit Monoclonal Antibody
Cat # M06634
M06634 renders with an IHC image of paraffin-embedded human kidney (catalog IHC image caption). It lists IHC and IF/ICC applications and human, mouse, and rat reactivity; no IF image is supplied (catalog applications/reactivity; catalog IF images).
Which to pick: Choose M06634 for paraffin-section tissue IHC based on its human kidney image; the fixative is unreported (catalog IHC image caption). For IF/ICC, M06634 lists both applications, though no IF image is supplied (catalog applications; catalog IF images). For human, mouse, or rat work, M06634 is the listed rabbit monoclonal option, clone FEH-18 (catalog reactivity; catalog host/clone).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P60866 (RS20_HUMAN, Small ribosomal subunit protein uS10).
- Human Protein Atlas. RPS20 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. RPS20 subcellular location (ICC-IF): Localized to the endoplasmic reticulum and cytosol..
- Human Protein Atlas. RPS20 antibody validation summary (1 antibodies).
- Biochemical and clinical effects of RPS20 expression in renal clear cell carcinoma. Oncology reports 2023 — PMC9773015.
- Ribosomal Proteins RPS11 and RPS20, Two Stress-Response Markers of Glioblastoma Stem Cells, Are Novel Predictors of Poor Prognosis in Glioblastoma Patients. PloS one 2015 — PMC4624638.
- Third-party punishment-like behavior in a rat model. Scientific reports 2024 — PMC11436845.
- New Pathogenic Germline Variants in Very Early Onset and Familial Colorectal Cancer Patients. Frontiers in genetics 2020 — PMC7541943.
- PubMed PMID:8479924 — UniProt-cited evidence.
- PubMed PMID:11875025 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RPS20 (Small ribosomal subunit protein uS10). Expected localisation General cytoplasmic staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: M06634 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Skeletal muscle. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human kidney tissue; fixative not specified (datasheet M06634); 2. Fixation: Image fixative and duration unreported (datasheet M06634); 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 monoclonal (clone FEH-18) anti-RPS20, 1:50 (datasheet M06634); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Skeletal myocytes are not detected (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope effect undetermined (UniProt); Regulation: Specific expression regulators unannotated (UniProt). Evidence: UniProt P60866, HPA tissue IHC · Approved, HPA ICC-IF · endoplasmic reticulum and cytosol., Datasheet M06634, PMC9773015, PMC4624638. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RPS20/immunohistochemistry-ihc-guide.