This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
RPS3 / Small ribosomal subunit protein uS3 · IHC design guide
Design Immunohistochemistry for RPS3
Plan chromogenic IHC for RPS3 in paraffin sections using the catalog antibody’s documented conditions (datasheet A01542-2). Compare staining with the generally cytoplasmic tissue pattern while accounting for the HPA assessment of uncertain reliability (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RPS3 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 | Predominantly cytoplasmic in tissue sections (HPA tissue IHC) | |
| Staining pattern | General cytoplasmic staining across tissues (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A01542-2) | |
| Positive control | Cerebellum+4 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 A01542-2) | |
| Caveat | Tissue staining has uncertain RNA concordance (HPA tissue IHC) | |
| Regulation | DNA damage can increase nuclear staining (UniProt) | |
| Isoform / epitope | Two isoforms; check antibody epitope coverage (UniProt) |
Section 1
Recommended RPS3 IHC & IF Protocols
Compare the catalog antibody’s citrate pH 6 IHC protocol (datasheet: A01542-2) with published RPS3 IHC protocols (PMC6393244; PMC9280127).
| Sample | Paraffin-embedded human tonsil tissues; fixative not specified (datasheet A01542-2) |
| Fixation | Image fixative and duration unreported (datasheet A01542-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A01542-2) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01542-2) |
| Primary antibody | Rabbit anti-RPS3, 0.5-1μg/ml (datasheet A01542-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01542-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01542-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RPS3-positive staining in purkinje cells of cerebellum (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate pH 6 retrieval (datasheet: A01542-2); PMC6393244 used pH 6, while PMC9280127 used pH 8.
Section 2
What Is the Expected RPS3 Staining Pattern?
RPS3 is mainly cytoplasmic in normal cells, with smaller nuclear amounts; it has no transmembrane segment (UniProt P23396). In paraffin sections, expect cytoplasmic staining in several cell types, including cerebellar Purkinje cells, cortical neurons, and esophageal squamous epithelial cells (HPA tissue IHC). Treat tissue comparisons as guides rather than fixed cutoffs: HPA rates the tissue IHC profile Uncertain because antibody staining and RNA expression show medium consistency (HPA tissue IHC).
| Clear cytoplasmic staining in Purkinje cells or cortical neurons, with readable tissue structure. | This fits HPA's High staining in those cells and its general cytoplasmic profile (HPA tissue IHC). Compare the signal with neighboring cells and the negative control on the same staining run (standard IHC practice). |
| Predominantly nuclear staining in otherwise normal tissue, with little cytoplasmic signal. | Review this pattern before calling it RPS3: normal cells contain mainly cytoplasmic RPS3, though some nuclear signal is possible (UniProt P23396). Nuclear accumulation can accompany apoptosis or DNA damage, so the compartment alone cannot establish an artefact (UniProt P23396). |
| Strong staining in adipocytes or skeletal muscle myocytes while expected positive cells are weak. | HPA reports RPS3 as Not detected in those cell types (HPA tissue IHC). Such staining raises concern for cross-reactivity or endogenous detection activity (standard IHC practice); HPA's Uncertain tissue reliability means these observations are checks, not absolute exclusions (HPA tissue IHC). |
| Diffuse color covers cells and extracellular spaces, obscuring cell boundaries. | That distribution is difficult to score as the general cytoplasmic expression reported by HPA (HPA tissue IHC). Review blocking, washing, detection reagents, and the negative control for nonspecific background or endogenous activity (standard IHC practice). |
| No signal in a section containing identifiable Purkinje cells, cortical neurons, or esophageal squamous epithelial cells. | These cells show High staining in HPA tissue IHC, so their absence of signal calls for a run-level check (HPA tissue IHC). Verify tissue identity, reagent performance, and the catalog antibody's IHC-P instructions before interpreting other negative cells (standard IHC practice). |
💡Expected RPS3 appearanceA persuasive positive result is predominantly cytoplasmic staining in identifiable High-staining cells such as Purkinje cells or cortical neurons (HPA tissue IHC; UniProt P23396); broad extracellular color or strong signal confined to HPA Not detected cell types warrants review (HPA tissue IHC; standard IHC practice).
| Tissue and cell identity (HPA tissue IHC) | HPA reports High staining in Purkinje cells, cortical and hippocampal neurons, cervical and salivary glandular cells, and esophageal and oral squamous epithelial cells; it reports Not detected in adipocytes and skeletal muscle myocytes (HPA tissue IHC). Score the named cells, not an entire tissue as uniformly positive. |
| Strength of the tissue evidence (HPA tissue IHC) | HPA describes general cytoplasmic expression but rates the tissue IHC profile Uncertain, citing medium consistency between antibody staining and RNA expression; its listed antibody, HPA063339, is also Uncertain for IHC (HPA tissue IHC; HPA antibodies). Use the pattern as an expectation requiring controls. |
| Cell state and compartment (UniProt P23396) | RPS3 is mainly cytoplasmic in normal cells, can translocate to nuclei during apoptosis or after DNA damage, and can accumulate in mitochondria with increased ROS (UniProt P23396). These reported changes qualify a compartment call; a nuclear or punctate result needs cellular context before interpretation. |
| Protein features versus assay behavior (UniProt P23396) | RPS3 has 2 isoforms, a KH type-2 domain, reported modified residues, and no transmembrane segment (UniProt P23396). These features do not identify the catalog antibody's epitope or establish isoform selectivity or retrieval requirements; assess those properties from antibody-specific validation. |
| Fixation evidence (HPA tissue IHC; UniProt P23396) | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells are blank. | A run or detection failure is possible when cells reported as High by HPA are unstained (HPA tissue IHC; standard IHC practice). | Confirm cell identity, inspect the run control, and check retrieval, antibody dilution, and detection against the catalog antibody's IHC-P instructions (standard IHC practice). No RPS3-specific retrieval condition is supplied. |
| Signal appears mostly nuclear. | Normal RPS3 is mainly cytoplasmic, but nuclear translocation is reported with apoptosis or DNA damage (UniProt P23396). | Examine cell morphology and cytoplasmic signal, then compare a suitable control section processed in the same run (standard IHC practice). Avoid assigning a cause to nuclear staining from compartment alone. |
| Strong color appears in HPA Not detected cells. | Cross-reactivity or endogenous detection activity may contribute (standard IHC practice); HPA's tissue profile is Uncertain (HPA tissue IHC). | Check a negative reagent control and compare staining in HPA High cells on the run (standard IHC practice; HPA tissue IHC). Treat any disagreement as a validation question rather than automatic proof of absence. |
| Diffuse background makes cytoplasmic scoring difficult. | Incomplete blocking, washing, or control of endogenous detection activity can produce background in chromogenic IHC (standard IHC practice). | Review the negative control, blocking and washing steps, and detection reagents; score only signal resolved within identifiable cells (standard IHC practice). HPA describes a general cytoplasmic pattern (HPA tissue IHC). |
| Staining differs markedly between tissue regions. | Cell composition matters: HPA lists High, Low, and Not detected results for different named cells (HPA tissue IHC). | Identify and score the relevant cell type in each region, then compare sections within the same run (standard IHC practice). Do not apply one intensity threshold to every cell type in the section. |
| Q: Should IF/ICC show the same compartment pattern? | HPA ICC-IF reports supported localization to the cytosol and endoplasmic reticulum, while the tissue IHC profile is Uncertain (HPA ICC-IF; HPA tissue IHC). | A: Use the cytosol and endoplasmic reticulum observation to interpret IF/ICC images (HPA ICC-IF); evaluate chromogenic paraffin sections against their own cell-specific IHC pattern and controls (HPA tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing RPS3 staining
A01542-2 · IHCIHC analysis of RPS3 using anti-RPS3 antibody (A01542-2). RPS3 was detected in paraffin-embedded section of human tonsil tissues. Heat media
A01542-2 · IHCIHC analysis of RPS3 using anti-RPS3 antibody (A01542-2). RPS3 was detected in paraffin-embedded section of human tonsil tissues. Heat media
M01542 · IHCImmunohistochemical analysis of paraffin-embedded human colon cancer, using RPS3 Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000149273-RPS3/tissue/cerebral+cortex#imid_17640835
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000149273-RPS3/tissue/colon#imid_17640769
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000149273-RPS3/tissue/liver#imid_17640770
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000149273-RPS3/tissue/kidney#imid_17640763
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA063339; green = RPS3, blue = nucleus, red = microtubules.
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA063339; green = RPS3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RPS3 IHC & IF
🧪Run cerebellum first and score Purkinje cells, where RPS3 staining is High (HPA: High in Purkinje cells). Use adipose tissue as a negative comparator, focusing on adipocytes (HPA: Not detected in adipocytes); on the cerebellum slide, use cells without specific signal to judge background without assigning a named cell type as RPS3-negative.
Positive control tissue: Cerebellum (Purkinje cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RPS3 in MCF-7, Rh30, U2OS, with annotated localisation: Endoplasmic reticulum (supported), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a concentration-matched rabbit IgG control matched to the primary antibody’s isotype and clonality where known (selected-SKU caption: rabbit primary), and an RPS3 knockout specimen or validated peptide-block control. Block endogenous peroxidase and check endogenous biotin background in cerebellum when using the caption’s biotin-based DAB detection (selected-SKU caption: biotinylated secondary, SABC and DAB).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption does not state a fixative (selected-SKU caption: fixative unreported). The reported IHC procedure uses heat retrieval in citrate buffer at pH 6 for 20 minutes; this supports starting with that condition but does not establish that retrieval is required (selected-SKU caption: citrate retrieval). Can IF/ICC be used instead? HPA reports supported cytosol and endoplasmic reticulum localization in ICC-IF images, but the supplied evidence does not establish that IF or frozen sections are easier than paraffin IHC; check cerebellar neuronal autofluorescence if using IF (HPA: subcellular ICC-IF).
HPA tissue IHC evidence for RPS3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — 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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | Follicle cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Chondrocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RPS3 IHC Tips
Use the matched paraffin-section protocol as the starting point, then interpret RPS3 staining by compartment and cell type (datasheet A01542-2; UniProt P23396; HPA tissue IHC).
What retrieval should I use if RPS3 staining is weak in paraffin sections?
Start with heat-mediated citrate retrieval at pH 6 for 20 minutes (datasheet A01542-2). The matched paraffin-section image used this condition before overnight incubation with 1 µg/ml antibody, so keep those variables together for the first comparison (datasheet A01542-2). If staining remains weak, compare a modestly longer retrieval or an alternative buffer on adjacent sections while holding detection and exposure constant (standard IHC practice). Watch for tissue damage and rising background, and judge improvement by cytoplasmic staining in the expected cells rather than by darker DAB alone (HPA tissue IHC; standard IHC practice).
Could fixation explain weak or uneven RPS3 staining?
The selected image documents paraffin embedding but gives no fixative, so RPS3-specific fixation sensitivity is unknown (datasheet A01542-2). Record the fixative and fixation interval for each specimen, then compare sections processed alike before changing retrieval or antibody concentration (standard IHC practice). Include a section with expected cytoplasmic staining in the same run to distinguish specimen handling from a failed staining run (HPA tissue IHC; standard IHC practice). Do not infer fixation tolerance from RPS3 localisation, its modified residues, or the HPA tissue pattern; none measures how this antibody performs after a particular fixation condition (UniProt P23396; HPA tissue IHC; datasheet A01542-2).
How should I assess RPS3 staining outside the cytoplasm?
Expect predominantly cytoplasmic staining in normal cells, with smaller nuclear amounts described for RPS3 (UniProt P23396). Nuclear accumulation can accompany apoptosis or DNA damage, while mitochondrial accumulation is reported with increased reactive oxygen species; these associations alone do not establish the state of a stained section (UniProt P23396). The HPA tissue profile describes general cytoplasmic expression, whereas its cell-imaging record supports cytosol and endoplasmic reticulum localisation (HPA tissue IHC; HPA subcellular). Score cytoplasmic and nuclear staining separately, inspect cell morphology, and confirm an unusual compartment pattern with appropriate controls before calling it biological redistribution (standard IHC practice).
Can this stain distinguish RPS3 isoforms or modified forms?
RPS3 has 2 annotated isoforms and multiple modified residues, including sites near its N terminus (UniProt P23396). The supplied antibody caption gives a working IHC concentration but no epitope map or isoform-specific validation, so a stained section cannot identify which isoform or modification produced the signal (datasheet A01542-2; UniProt P23396). Its KH type-2 domain spans residues 21–92, but that domain annotation does not locate this antibody's binding site (UniProt P23396; datasheet A01542-2). For an isoform or modification claim, obtain epitope information and test a suitably specific reagent or orthogonal assay alongside the IHC pattern (standard IHC practice).
How should I investigate RPS3 localisation by multiplex IF?
Plan IF as a separate validation experiment: the selected antibody evidence describes chromogenic staining of paraffin sections, not an IF result for this antibody (datasheet A01542-2). Pair RPS3 with a marker identifying the expected cell population, such as the glandular cells reported as strongly stained in salivary gland, and choose spectrally separated fluorophores after checking tissue autofluorescence (HPA tissue IHC; standard IF practice). RPS3 has no annotated transmembrane segment, but this antibody's epitope is unspecified; establish intracellular epitope access with a controlled permeabilisation comparison (UniProt P23396; datasheet A01542-2; standard IF practice). Include single-label and secondary-only controls before interpreting apparent overlap or nuclear signal (standard IF practice).
How can I separate diffuse RPS3 staining from chromogenic background?
The matched paraffin-section workflow used 10% goat-serum block, a biotinylated secondary, streptavidin–biotin detection, and DAB (datasheet A01542-2). Apply a peroxidase block and compare no-primary and detection-only controls to identify enzyme or detection background in the chromogenic workflow (standard IHC practice). If diffuse colour persists, check whether background follows tissue edges or appears in cells expected to have little staining, such as adipocytes in the HPA profile (HPA tissue IHC; standard IHC practice). Titrate detection and primary incubation separately, preserving the matched 1 µg/ml overnight condition as the reference rather than treating every darker cell as positive (datasheet A01542-2; standard IHC practice).
What should I score when comparing RPS3 IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and compartment before viewing groups, because RPS3 is generally cytoplasmic but may redistribute to the nucleus under particular conditions (HPA tissue IHC; UniProt P23396; standard IHC practice). Within each region, record the percentage of positive cells and an intensity-based H-score, with cytoplasmic and nuclear results kept separate (standard IHC practice). Normalise positive-cell counts to the number of evaluable cells, or report positive-cell density per mm² of viable annotated tissue when cell counts are impractical (standard IHC practice). Use identical retrieval, DAB development, imaging settings, and scoring thresholds across sections, and exclude damaged or necrotic areas (standard IHC practice).
When is an unexpected RPS3-positive cell or compartment credible?
A credible pattern should localise within intact cells and agree with an independently identified cell population; HPA reports high staining in several neuronal, glandular, and squamous epithelial populations (HPA tissue IHC; standard IHC practice). Predominantly cytoplasmic staining fits the reported baseline, while strong nuclear staining merits a check for apoptosis or DNA-damage context rather than an automatic positive call (UniProt P23396). Staining confined to cut edges, necrotic areas, or no-primary controls suggests a processing or detection artefact (standard IHC practice). Interpret unexpected positives cautiously because HPA rates its tissue IHC reliability Uncertain, and check compartment, morphology, and controls together (HPA tissue IHC; standard IHC practice).
Boster reagents
Best RPS3 / Small ribosomal subunit protein uS3 IHC Antibodies
Anti-RPS3 antibodies have paraffin-section IHC images from human tonsil, placenta and colon cancer, and mouse brain (catalog image captions). Both list IF/ICC applications, but neither has an IF image (catalog).
Real IHC data
Anti-RPS3 Antibody ®
Cat # A01542-2
Real IHC data
Anti-RPS3/Ribosomal Protein S3 Rabbit Monoclonal Antibody
Cat # M01542
A01542-2 lists IHC and IF/ICC applications and human, mouse and rat reactivity; its IHC captions show paraffin sections of human tonsil and placenta and mouse brain (catalog). M01542 lists the same applications and species reactivity; its IHC caption shows paraffin-embedded human colon cancer (catalog).
Which to pick: For tissue IHC, choose A01542-2 for its documented citrate pH 6 retrieval on paraffin sections; choose M01542 if its rabbit monoclonal format and human colon cancer example suit the study (A01542-2 image captions; M01542 catalog and image caption). Both list IF/ICC, though neither supplies an IF image; the listed IF dilutions are 2 μg/ml for A01542-2 and 1:50 for M01542 (catalog). Both list human, mouse and rat reactivity, but A01542-2 has the mouse tissue IHC example; neither product’s IHC caption reports the fixative (catalog; image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P23396 (RS3_HUMAN, Small ribosomal subunit protein uS3).
- Human Protein Atlas. RPS3 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. RPS3 subcellular location (ICC-IF): Localized to the endoplasmic reticulum and cytosol..
- Human Protein Atlas. RPS3 antibody validation summary (1 antibodies).
- RNA-binding protein RPS3 contributes to hepatocarcinogenesis by post-transcriptionally up-regulating SIRT1. Nucleic acids research 2019 — PMC6393244.
- IKKβ phosphorylation regulates RPS3 nuclear translocation and NF-κB function during infection with Escherichia coli strain O157:H7. Nature immunology 2011 — PMC3062687.
- PELI1: key players in the oncogenic characteristics of pancreatic Cancer. Journal of experimental & clinical cancer research : CR 2024 — PMC10962118.
- RPS3 Promotes the Metastasis and Cisplatin Resistance of Adenoid Cystic Carcinoma. Frontiers in oncology 2022 — PMC9280127.
- PubMed PMID:2129557 — UniProt-cited evidence.
- PubMed PMID:1712897 — UniProt-cited evidence.
- PubMed PMID:11875025 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RPS3 (Small ribosomal subunit protein uS3). Expected localisation Predominantly cytoplasmic in tissue sections (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A01542-2). Validated IHC antibody: A01542-2 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissues; fixative not specified (datasheet A01542-2); 2. Fixation: Image fixative and duration unreported (datasheet A01542-2); 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: Citrate pH 6 , 20 min (datasheet A01542-2); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01542-2); 8. Primary antibody: Rabbit anti-RPS3, 0.5-1μg/ml (datasheet A01542-2); 9. Primary incubation: Overnight at 4 °C (datasheet A01542-2). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01542-2); Caveat: Tissue staining has uncertain RNA concordance (HPA tissue IHC); Isoform / epitope: Two isoforms; check antibody epitope coverage (UniProt); Regulation: DNA damage can increase nuclear staining (UniProt). Evidence: UniProt P23396, HPA tissue IHC · Uncertain, HPA ICC-IF · endoplasmic reticulum and cytosol., Datasheet A01542-2, PMC6393244, PMC9280127. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RPS3/immunohistochemistry-ihc-guide.