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RPS13 / Small ribosomal subunit protein uS15 · IHC design guide
Design Immunohistochemistry for RPS13
RPS13 shows cytoplasmic staining in several tissues, but the tissue IHC evidence is rated uncertain (HPA tissue IHC). This guide helps plan paraffin-section controls and interpret staining using the catalog antibody’s documented IHC conditions (datasheet A06221-3).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RPS13 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 several tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in several tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06221-3) | |
| Positive control | Kidney+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 | Low staining–RNA consistency; verify the pattern (HPA tissue IHC) | |
| Regulation | No expression regulator specified (UniProt) | |
| Isoform / epitope | 0 annotated isoforms; epitope effects unknown (UniProt) |
Section 1
Recommended RPS13 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A06221-3) is accompanied by one published gastric tissue IHC protocol (PMC3822796).
| Sample | Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet A06221-3) |
| Fixation | Image fixative and duration unreported (datasheet A06221-3); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A06221-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06221-3) |
| Primary antibody | Rabbit anti-RPS13, 2-5 μg/ml (datasheet A06221-3) |
| Primary incubation | Overnight at 4 °C (datasheet A06221-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06221-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RPS13-positive staining in cells in glomeruli of kidney (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet: A06221-3). The published protocol does not report retrieval (PMC3822796).
Section 2
What Is the Expected RPS13 Staining Pattern?
RPS13 is a cytoplasmic and nucleolar ribosomal protein with no transmembrane segment (UniProt P62277: localization and topology). In paraffin IHC, expect cytoplasmic staining in several tissues, including glomerular cells, germinal center cells, splenic white pulp cells, and stomach glandular cells (HPA: tissue IHC). Treat the pattern as provisional: HPA rates tissue IHC reliability Uncertain because antibody staining has low consistency with RNA expression (HPA: reliability).
| Distinct cytoplasmic staining in glomerular cells or germinal center cells, with limited background (HPA: High in these cells). | This fits the reported tissue IHC distribution and UniProt cytoplasmic localization (HPA: tissue IHC; UniProt P62277: localization). It supports an interpretable result, although HPA rates the tissue pattern Uncertain (HPA: reliability). |
| Staining is confined to an unexpected compartment while cytoplasmic signal is absent (HPA: cytoplasmic tissue IHC). | Suspect a localization or detection artifact and review controls. UniProt also lists the nucleolus, so nucleolar staining alone cannot establish an artifact (UniProt P62277: localization). HPA tissue IHC chiefly reports cytoplasmic expression (HPA: profile). |
| Strong staining appears in adipocytes or bronchial respiratory epithelial cells (HPA: Not detected in these cells). | Consider cross-reactivity or endogenous detection activity; check a no-primary control and compare staining with the expected cells. These are reported negatives for the sampled cell types, not proof that every cell in either tissue lacks RPS13 (HPA: tissue IHC). |
| Color covers tissue and empty spaces without a recognizable cellular pattern. | Interpret this as diffuse background, not a positive RPS13 result. Check the no-primary control, blocking, washing, and chromogenic detection as general IHC troubleshooting; HPA's uncertain tissue reliability makes weak patterns especially difficult to assign (HPA: reliability). |
| No signal appears in glomerular cells, germinal center cells, or stomach glandular cells (HPA: High in these cells). | First suspect an unsuccessful IHC run or insufficient detection and verify the positive-control section, retrieval, and antibody conditions. HPA's High designations identify useful comparison cells but do not guarantee staining in every specimen (HPA: tissue IHC; reliability Uncertain). |
💡Expected RPS13 appearanceCall a result provisionally positive when cytoplasmic staining is clear and relatively strong in HPA High cells, such as glomerular or germinal center cells; diffuse color or strong signal in HPA Not detected cell types warrants control review (HPA: tissue IHC; reliability Uncertain).
| Cell-type choice and evidence strength | Glomerular cells, germinal center cells, splenic white pulp cells, and stomach glandular cells are reported High; lung macrophages and cardiomyocytes are Medium (HPA: tissue IHC). Choose a reported High population for comparison, while retaining the Uncertain reliability caveat (HPA: reliability). |
| Compartment and topology | RPS13 is assigned to the cytoplasm and nucleolus and has no transmembrane segment (UniProt P62277: localization and topology). HPA tissue IHC summarizes cytoplasmic expression; assess compartment and cell identity together before interpreting nuclear or diffuse color (HPA: tissue IHC). |
| Maturation and variants | UniProt lists a 2–151 chain, no signal peptide or propeptide, and 0 annotated isoforms (UniProt P62277: processing and isoforms). These annotations provide no basis here for predicting a secreted staining pattern or different isoform-specific tissue patterns. |
| Antibody validation | The listed antibody HPA005985 is rated Uncertain for IHC and Approved for ICC (HPA: antibody validation). Neither rating upgrades the HPA tissue pattern to a confirmed IHC result; compare slide morphology and controls before attributing an unexpected pattern to RPS13. |
| IF/ICC Q&A: should its pattern match paraffin IHC? | HPA reports endoplasmic reticulum localization in ICC-IF images from HEK293 and U2OS, while tissue IHC is chiefly cytoplasmic (HPA: subcellular; tissue IHC). UniProt also lists the nucleolus (UniProt P62277: localization). Interpret each assay within its own evidence and controls. |
| Situation | Likely cause | Next action |
|---|---|---|
| High cells lack staining while the section is otherwise readable (HPA: High in glomerular or germinal center cells). | The IHC workflow or detection may have failed; the HPA High designation does not guarantee every specimen will stain (HPA: reliability Uncertain). | Review the run's positive control, primary-antibody conditions, antigen retrieval, and detection reagents using general paraffin IHC practice. |
| Adipocytes stain strongly (HPA: Not detected in adipocytes). | Cross-reactivity or endogenous detection activity is possible; HPA tissue IHC has Uncertain reliability (HPA: tissue IHC; reliability). | Inspect the no-primary control, assess cellular morphology, and compare with a reported High cell population on a separately controlled section (HPA: tissue IHC). |
| Diffuse chromogen obscures cell boundaries. | Nonspecific binding, residual detection reagent, or excessive color development can cause background in chromogenic IHC (general IHC practice). | Check no-primary staining, blocking, washes, and development time; score RPS13 only where cellular localization remains distinguishable (general IHC practice). |
| Signal appears only in nuclei or nucleoli. | UniProt includes nucleolar localization, whereas HPA describes chiefly cytoplasmic tissue IHC (UniProt P62277: localization; HPA: profile). | Check counterstain and detection controls, then describe the observed compartment without calling it confirmed tissue IHC localization (HPA: reliability Uncertain). |
| Different cell types stain with similar intensity across the section. | A broad signal may obscure the reported High, Medium, and Not detected cell-type distinctions (HPA: tissue IHC). | Compare defined cell populations and the no-primary control; adjust general IHC detection conditions if background prevents cell-level assessment. |
| IHC and ICC-IF images suggest different compartments. | HPA describes cytoplasmic tissue IHC but endoplasmic reticulum localization in ICC-IF; its tissue IHC reliability is Uncertain (HPA: tissue IHC; subcellular; reliability). | Report the assay and cell context with each observation. Use the ICC-IF result as context, not as a required paraffin IHC pattern (HPA: tissue IHC; subcellular). |
Experiment evidence
IHC & IF images showing RPS13 staining
A06221-3 · IHCIHC analysis of RPS13 using anti-RPS13 antibody (A06221-3). RPS13 was detected in a paraffin-embedded section of human colon adenocarcinoma
A06221-3 · IHCIHC analysis of RPS13 using anti-RPS13 antibody (A06221-3). RPS13 was detected in a paraffin-embedded section of human endometrial carcinoma
A06221-1 · IHCImmunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue, using RPS13 Antibody. The picture on the right is blocked
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000110700-RPS13/tissue/cerebral+cortex#imid_1848448
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000110700-RPS13/tissue/colon#imid_1844766
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000110700-RPS13/tissue/liver#imid_1844759
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000110700-RPS13/tissue/kidney#imid_1844768
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA005985; green = RPS13, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA005985; green = RPS13, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RPS13 IHC & IF
🧪Run tonsil first: germinal center cells should stain (HPA: High in tonsil germinal center cells). Use adipose tissue as the negative comparator, assessing its adipocytes specifically (HPA: Not detected in adipocytes); within the tonsil slide, cells judged negative should show counterstain without specific chromogen, confirmed against the no-primary control (standard IHC practice).
Positive control tissue: Kidney (Cells in glomeruli, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RPS13 in HEK293, U2OS, with annotated localisation: Endoplasmic reticulum (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a concentration-matched rabbit IgG isotype control matched to the primary antibody’s clonality, and an RPS13-knockout sample as a biological negative (selected SKU caption: rabbit primary; standard IHC controls). In tonsil, quench endogenous peroxidase and check the no-primary slide for residual DAB signal in inflammatory cells (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A06221-3 paraffin-section caption does not state the fixative (selected SKU caption). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required (selected SKU caption). Neither frozen sections nor IF is established as easier; for tonsil IHC, check inflammatory-cell peroxidase background before scoring (standard IHC practice).
HPA tissue IHC evidence for RPS13
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Kidney | Cells in glomeruli | High | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
| Spleen | Cells in white pulp | High | Protein (IHC) | HPA → |
| Stomach | Glandular cells | High | Protein (IHC) | HPA → |
| Tonsil | Germinal center cells | High | 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 → |
| Cervix | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RPS13 IHC Tips
Troubleshoot RPS13 staining in paraffin sections by checking retrieval, compartment, controls, and scoring before interpreting differences between specimens.
What retrieval should I start with for RPS13 in paraffin sections?
Start with heat mediated retrieval in EDTA at pH 8.0 (datasheet A06221-3). The catalog antibody detected RPS13 in a paraffin section of human colon adenocarcinoma after this retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet A06221-3). If staining is weak, compare a shorter and longer heating interval on matched sections while keeping the EDTA buffer and detection conditions constant (standard IHC practice). Include a known positive section and inspect tissue preservation, because stronger retrieval can increase background or damage morphology (standard IHC practice).
Could fixation explain weak or uneven RPS13 staining?
Target specific fixation sensitivity is unknown: the selected paraffin section caption does not state its fixative (datasheet A06221-3). Record fixation method and duration for each specimen, then compare sections processed together with the same EDTA pH 8.0 retrieval and antibody incubation (datasheet A06221-3; standard IHC practice). If staining differs across blocks, examine morphology and use a matched positive control before attributing the difference to RPS13 abundance (standard IHC practice). Do not infer fixation tolerance from the reported tissue staining pattern or from RPS13 localisation and modifications; neither establishes a fixation response (HPA: tissue IHC; UniProt P62277).
Which compartments should show convincing RPS13 staining?
Assess cytoplasmic staining first: tissue IHC describes cytoplasmic expression in several tissues, while UniProt also places RPS13 in the nucleus and nucleolus (HPA: tissue IHC; UniProt P62277). RPS13 has no transmembrane segment, so a sharp cell surface outline alone is discordant with its annotated topology (UniProt P62277). HPA subcellular imaging instead reports an approved endoplasmic reticulum location, creating a compartment discrepancy that warrants caution when reading paraffin sections (HPA: subcellular). Compare the suspected signal with adjacent morphology, a positive section, and a no primary control before assigning a compartment (standard IHC practice).
How can epitope uncertainty affect RPS13 IHC?
The record lists 0 isoforms and a processed chain spanning residues 2–151, but supplies no antibody epitope coordinates (UniProt P62277; datasheet A06221-3). Several modifications occur near the amino terminus, including alternatives at residue 27 and phosphorylation at residue 30; their effect on this antibody is unknown (UniProt P62277). If staining changes between specimens, compare retrieval and fixation records before invoking epitope masking by a particular modification (standard IHC practice). Confirm unexpected patterns with an independently validated antibody against a mapped, distinct epitope if one is available, and retain the same tissue controls (standard IHC practice).
How should I investigate RPS13 by IF when tissue IHC is ambiguous?
Use IF as a separate validation experiment, because this page's selected antibody evidence is paraffin section chromogenic IHC (datasheet A06221-3). Multiplex RPS13 with a marker for the expected cell population, such as a germinal center cell marker when examining the HPA reported lymph node signal (HPA: High in lymph node germinal center cells; standard IF practice). Choose a far red fluorophore and include an unstained tissue control to assess autofluorescence in other channels (standard IF practice). Permeabilise cells for intracellular access, then inspect cytoplasm and nucleoli; RPS13 lacks a transmembrane segment and has annotated cytoplasmic and nucleolar locations (UniProt P62277; standard IF practice).
How can I reduce diffuse brown staining without losing RPS13 signal?
First compare the test section with a no primary control, and block endogenous peroxidase before HRP and DAB detection (standard IHC practice). The selected caption used 10% goat serum, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase conjugated secondary for 30 minutes at 37°C (datasheet A06221-3). If background persists, adjust primary concentration or wash stringency on matched sections while retaining the EDTA pH 8.0 retrieval and a positive control (datasheet A06221-3; standard IHC practice). Exclude edge staining, deposits, and necrotic areas from any RPS13 interpretation (standard IHC practice).
How should I score RPS13 across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then use an H-score based on staining intensity and the percentage of positive cells (standard IHC practice). Record the percentage positive separately, and normalise counts or stained area to the number of evaluable cells or viable tissue area in mm² (standard IHC practice). Keep retrieval, chromogen development, imaging, and scoring thresholds consistent across the comparison set (standard IHC practice). Report compartment specific results because tissue IHC describes cytoplasmic expression, while UniProt also annotates nuclear and nucleolar RPS13; do not pool these patterns without stating the rule (HPA: tissue IHC; UniProt P62277).
When is an apparent RPS13 positive result convincing?
A convincing result has reproducible intracellular staining in the expected cell population and is absent from the no primary control (standard IHC practice). HPA reports high staining in lymph node germinal center cells and stomach glandular cells, but rates its tissue IHC reliability Uncertain because staining and RNA expression show low consistency (HPA: tissue IHC). Treat isolated membrane outlines cautiously because RPS13 has no transmembrane segment; compare unusual compartment patterns with its cytoplasmic, nuclear, and nucleolar annotations (UniProt P62277). Reject edge effects, necrotic staining, and residual endogenous peroxidase signal before interpreting a biological difference (standard IHC practice).
Boster reagents
Best RPS13 / Small ribosomal subunit protein uS15 IHC Antibodies
Two anti-RPS13 antibodies have paraffin-section IHC images from human carcinoma tissue (catalog captions). IF/ICC is listed for A06221-1, with no IF image supplied (catalog applications and images).
Real IHC data
Anti-RPS13 Antibody ®
Cat # A06221-3
Real IHC data
Anti-RPS13/Ribosomal Protein S13 Antibody
Cat # A06221-1
A06221-3 has IHC images from human colon adenocarcinoma, endometrial carcinoma, and prostate cancer paraffin sections (A06221-3 image captions). A06221-1 has an IHC image from a human breast carcinoma paraffin section, with peptide-blocked staining shown alongside it (A06221-1 image caption).
Which to pick: For human tissue IHC, choose A06221-3 if its documented EDTA retrieval at pH 8.0 and 2 μg/ml primary incubation suit the experiment (A06221-3 image captions); its caption identifies paraffin sections but does not report the fixative. For IF/ICC or mouse and rat samples, choose the polyclonal A06221-1 because those applications and species are listed (A06221-1 catalog); its IHC caption documents a human paraffin section but does not report the fixative, and no IF image is supplied (A06221-1 image caption and catalog images).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P62277 (RS13_HUMAN, Small ribosomal subunit protein uS15).
- Human Protein Atlas. RPS13 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. RPS13 subcellular location (ICC-IF): Localized to the endoplasmic reticulum..
- Human Protein Atlas. RPS13 antibody validation summary (1 antibodies).
- Human ribosomal protein S13 promotes gastric cancer growth through down-regulating p27(Kip1). Journal of cellular and molecular medicine 2011 — PMC3822796.
- PubMed PMID:8332508 — UniProt-cited evidence.
- PubMed PMID:8920921 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RPS13 (Small ribosomal subunit protein uS15). Expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06221-3). Validated IHC antibody: A06221-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in glomeruli); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet A06221-3); 2. Fixation: Image fixative and duration unreported (datasheet A06221-3); 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: EDTA pH 8.0 (datasheet A06221-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06221-3); 8. Primary antibody: Rabbit anti-RPS13, 2-5 μg/ml (datasheet A06221-3); 9. Primary incubation: Overnight at 4 °C (datasheet A06221-3). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Low staining–RNA consistency; verify the pattern (HPA tissue IHC); Isoform / epitope: 0 annotated isoforms; epitope effects unknown (UniProt); Regulation: No expression regulator specified (UniProt). Evidence: UniProt P62277, HPA tissue IHC · Uncertain, HPA ICC-IF · endoplasmic reticulum., Datasheet A06221-3, PMC3822796. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RPS13/immunohistochemistry-ihc-guide.