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RPS12 / Small ribosomal subunit protein eS12 · IHC design guide
Design Immunohistochemistry for RPS12
Plan chromogenic RPS12 IHC in paraffin sections using the catalog antibody's documented workflow (datasheet A01040-3). Expect a general cytoplasmic pattern, with high staining in selected epithelial and lymph node germinal center cells; interpret intensity in light of medium consistency with RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RPS12 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 tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in epithelial and germinal center cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01040-3) | |
| Positive control | Bronchus+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 | Staining has medium consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Low tissue specificity; no regulator reported (HPA tissue IHC; UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain spans residues 2–132 (UniProt) |
Section 1
Recommended RPS12 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet: A01040-3); one published RPS12 IHC protocol uses citrate retrieval (PMC12291262).
| Sample | Paraffin-embedded human thyroid cancer tissue; fixative not specified (datasheet A01040-3) |
| Fixation | Image fixative and duration unreported (datasheet A01040-3); 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 A01040-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01040-3) |
| Primary antibody | Rabbit anti-RPS12, 1:50 recommended; image 1:100 (datasheet A01040-3) |
| Primary incubation | Overnight at 4 °C (datasheet A01040-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01040-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RPS12-positive staining in basal cells of bronchus (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 (datasheet: A01040-3). If optimizing retrieval, compare the published citrate condition (PMC12291262).
Section 2
What Is the Expected RPS12 Staining Pattern?
RPS12 is a cytoplasmic protein that also localizes to the nucleus and nucleolus (UniProt P25398). It has no transmembrane segment (UniProt P25398 topology). In paraffin tissue sections, expect mainly cytoplasmic staining, including strong signal in bronchial basal cells and several epithelial and glandular cell populations (HPA tissue IHC: High). The tissue IHC profile is Approved, with medium consistency between antibody staining and RNA expression (HPA tissue IHC: reliability).
| Strong cytoplasmic staining in bronchial basal cells or duodenal glandular cells, with recognizable cell boundaries. | This fits the reported high staining in those cell populations (HPA tissue IHC: High in bronchial basal cells and duodenal glandular cells). Judge intensity within the named cells; neighboring cells need not stain equally. Cytoplasmic predominance also fits the reported tissue profile (HPA tissue IHC: general cytoplasmic expression). |
| Nuclear or nucleolar signal accompanies cytoplasmic staining; alternatively, staining is predominantly membranous or extracellular. | Nuclear and nucleolar localization is plausible (UniProt P25398: nucleus, nucleolus). A predominantly membranous or extracellular pattern is inconsistent with the reported cytoplasmic localization and absence of a transmembrane segment (HPA tissue IHC: cytoplasmic; UniProt P25398 topology). Check controls before calling it an artefact. |
| Strong staining appears in adipocytes or cardiomyocytes while a reported high staining population is weak. | Those specific cell populations are reported as not detected, whereas several epithelial and glandular populations stain strongly (HPA tissue IHC: adipocytes and cardiomyocytes Not detected; bronchial basal cells High). Consider cross-reactivity or endogenous detection activity; the mismatch alone does not identify its cause. |
| Brown deposit spreads across stroma, empty spaces, or many unrelated cell types, obscuring cytoplasm. | A diffuse deposit is hard to reconcile with cell-resolved cytoplasmic staining (HPA tissue IHC: general cytoplasmic expression). Review a no-primary control for detection background, then assess blocking and washes (general chromogenic IHC practice). Do not score deposit outside identifiable cells as RPS12-positive. |
| No signal is visible in bronchial basal cells or cervical squamous epithelial cells. | Both are reported high staining populations, so absence of signal calls for a technical check (HPA tissue IHC: High in bronchial basal cells and cervical squamous epithelial cells). The HPA profile has medium consistency with RNA data; a single blank section does not establish true absence (HPA tissue IHC: reliability). |
💡Expected RPS12 appearanceCall a convincing positive result cell-resolved cytoplasmic staining in a reported high staining population, such as bronchial basal cells (HPA tissue IHC: High; general cytoplasmic expression); treat predominantly membranous or extracellular deposit as suspect (UniProt P25398 topology; HPA tissue IHC: cytoplasmic).
| Choice of comparison cells | Use named cells within a tissue: duodenal glandular cells are reported High, while adipocytes are Not detected (HPA tissue IHC). A negative call for cholangiocytes must not be extended to every cell in liver (HPA tissue IHC: cholangiocytes Not detected). |
| Strength of tissue evidence | The tissue IHC profile is Approved but has medium consistency with RNA expression (HPA tissue IHC: reliability). Interpret an unexpected result alongside cell identity, localization, and controls; do not treat an HPA staining category as an absolute threshold for every specimen. |
| Antibody validation | HPA006124 is listed as IHC Approved; HPA006365 is listed as ICC Supported, with no IHC status supplied for it (HPA antibodies). The supplied validation record does not label the tissue result Enhanced (HPA antibodies). Keep IHC and ICC evidence separate. |
| Protein localization and processing | Cytoplasmic and nucleolar signal can fit RPS12 biology (UniProt P25398: cytoplasm, nucleus, nucleolus). No transmembrane segment or signal peptide is annotated, and the listed chain spans residues 2–132 (UniProt P25398 topology and processing); a surface-only pattern warrants scrutiny. |
| IF/ICC Q&A: What pattern is expected? | Mainly cytosolic fluorescence, with additional Golgi apparatus and vesicle localization, is reported in ICC-IF (HPA subcellular: Cytosol Supported; Golgi apparatus and Vesicles Approved). This is localization context for IF/ICC; the IHC interpretation here relies on the tissue IHC record. |
| Situation | Likely cause | Next action |
|---|---|---|
| The reported high staining cells are blank. | The run may have lost signal during retrieval, antibody incubation, or detection (general chromogenic IHC practice); these data do not establish RPS12-specific fixation or retrieval sensitivity. | Check a reported high staining cell population on the same run, such as bronchial basal cells (HPA tissue IHC: High). Review the catalog antibody's IHC-P instructions and run controls before changing a condition (general IHC practice). |
| Nearly every area of the section turns brown. | Endogenous detection activity or nonspecific reagent binding can produce widespread color (general chromogenic IHC practice). This appearance cannot be assigned to RPS12 from color alone. | Compare a no-primary control and inspect the detection reagent and blocking steps (general chromogenic IHC practice). Reassess whether identifiable high staining cells retain stronger cytoplasmic signal (HPA tissue IHC: general cytoplasmic expression). |
| Strong color appears in adipocytes or cardiomyocytes. | Those named cells are reported Not detected (HPA tissue IHC). Cross-reactivity, background, or specimen variation could explain a discrepancy; the HPA tissue profile has medium consistency (HPA tissue IHC: reliability). | Confirm cell identity, compare a no-primary control, and inspect a reported high staining population in the same run (general IHC practice; HPA tissue IHC: High in bronchial basal cells). |
| Only cell edges or extracellular material stain. | That distribution conflicts with the reported cytoplasmic tissue profile and no-transmembrane topology (HPA tissue IHC: general cytoplasmic expression; UniProt P25398 topology). Nonspecific deposit is possible. | Review the counterstain and cell boundaries, compare the no-primary control, and reassess primary antibody concentration using its IHC-P instructions (general chromogenic IHC practice). |
| The section has widespread haze but little cell-resolved staining. | Excess primary reagent, incomplete blocking, or inadequate washing can obscure localization (general chromogenic IHC practice). Haze does not match the reported general cytoplasmic pattern (HPA tissue IHC). | Check blocking, washes, and detection exposure against the validated IHC-P workflow (general IHC practice). Score only interpretable cells after background is controlled. |
| Nuclear staining appears stronger than expected. | RPS12 can localize to the nucleus and nucleolus (UniProt P25398), while the reported tissue IHC profile is generally cytoplasmic (HPA tissue IHC). Nuclear signal alone is therefore insufficient to label a section false positive. | Examine whether cytoplasmic staining remains in reported high staining cells and whether the no-primary control is clean (HPA tissue IHC: High; general IHC practice). Record nuclear and cytoplasmic compartments separately. |
Experiment evidence
IHC & IF images showing RPS12 staining
A01040-3 · IHCIHC analysis of RPS12 using anti-RPS12 antibody (A01040-3). RPS12 was detected in a paraffin-embedded section of human thyroid cancer tissue
A01040-3 · IHCIHC analysis of RPS12 using anti-RPS12 antibody (A01040-3). RPS12 was detected in a paraffin-embedded section of human liver cancer tissue.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000112306-RPS12/tissue/cerebral+cortex#imid_2100936
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000112306-RPS12/tissue/colon#imid_2100862
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000112306-RPS12/tissue/liver#imid_2100863
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000112306-RPS12/tissue/kidney#imid_2100864
HPA · ICC-IF SK-MEL-30Human Protein Atlas immunofluorescence, SK-MEL-30, antibody HPA006365; green = RPS12, blue = nucleus, red = microtubules.
HPA · ICC-IF SK-MEL-30Human Protein Atlas immunofluorescence, SK-MEL-30, antibody HPA006365; green = RPS12, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RPS12 IHC & IF
🧪Run bronchus first and require staining in basal cells (HPA: High in bronchus basal cells); use skeletal muscle myocytes as the negative tissue (HPA: Not detected in skeletal muscle myocytes). Internal negative cells on the bronchus slide should show only background chromogen, but the supplied HPA rows do not establish a specific bronchial cell population as target negative (HPA: bronchus basal cells High).
Positive control tissue: Bronchus (Basal cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RPS12 in SK-MEL-30, U-251MG, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; rabbit IgG matched to the primary antibody’s isotype, clonality, and concentration; and, if available, an RPS12-knockout specimen or cognate-peptide block (standard IHC practice; hero caption: rabbit anti-RPS12 antibody). Quench endogenous peroxidase and inspect inflammatory cells for residual signal before interpreting DAB staining in bronchus (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A01040-3 paraffin-section caption does not state the fixative (hero caption: fixative not stated). The reported IHC conditions use heat retrieval in EDTA at pH 8.0 and primary antibody at 1:100 overnight at 4°C; the caption does not establish whether retrieval is required (hero caption). HPA has ICC-IF images showing mainly cytosolic signal, but the supplied evidence does not establish whether frozen sections or IF are easier than paraffin IHC; in bronchus, check for endogenous peroxidase signal from inflammatory cells (HPA: cytosol supported; standard chromogenic IHC practice).
HPA tissue IHC evidence for RPS12
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Bronchus | Basal cells | High | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RPS12 IHC Tips
These questions address RPS12 staining in paraffin sections with chromogenic IHC; the immunofluorescence entry covers the separate IF/ICC application.
What should I change when RPS12 staining is weak after antigen retrieval?
Begin with heat-mediated retrieval in EDTA at pH 8.0 for paraffin RPS12 IHC (datasheet A01040-3). If staining is weak, check that sections were fully deparaffinised and that the retrieval solution covered them throughout heating (standard IHC practice). Compare the suspect run with a concurrently processed positive section; high staining in bronchial basal cells is a documented reference pattern (HPA tissue IHC: High in bronchial basal cells). The selected image used 1:100 primary antibody overnight at 4°C, so avoid attributing weak signal to retrieval before checking primary incubation and detection (caption A01040-3). Keep retrieval conditions identical across sections intended for scoring (standard IHC practice).
Could fixation explain variable RPS12 staining between paraffin blocks?
Target-specific RPS12 sensitivity to fixation is unknown because the selected paraffin-section caption does not state a fixative (caption A01040-3). Record each block's fixative and processing history before comparing staining, since fixation and processing can alter epitope accessibility in IHC (standard IHC practice). Process a reference section alongside suspect blocks using the same EDTA pH 8.0 retrieval and detection workflow (datasheet A01040-3; standard IHC practice). If staining varies, assess tissue preservation and retrieval consistency before assigning a biological difference (standard IHC practice). RPS12 topology and modified-residue annotations do not establish a fixation effect (UniProt P25398 topology and modified residues).
How should I assess cytoplasmic, nucleolar, or punctate RPS12 staining?
Expect a predominantly cytoplasmic tissue pattern, while keeping nucleolar localisation biologically plausible for RPS12 (HPA tissue IHC: general cytoplasmic expression; UniProt P25398 subcellular location). Its participation in the nucleolar small subunit processome supports examining nucleoli carefully, but does not make every nuclear DAB deposit specific (UniProt P25398 function; standard IHC interpretation). Compare cytoplasmic and nucleolar staining within intact cells and across similarly processed sections (standard IHC practice). Golgi and vesicular locations have been reported by ICC/IF, so puncta merit comparison with the overall cellular pattern rather than automatic rejection (HPA subcellular: Golgi apparatus and vesicles). Check punctate signal against counterstain, tissue integrity, and a detection control (standard IHC practice).
Can an isoform or membrane barrier explain unexpected RPS12 staining?
The supplied record lists 0 isoforms, no transmembrane segment, and a chain spanning residues 2–132 (UniProt P25398 isoforms, topology and processing). These annotations provide no basis for assigning different IHC patterns to named RPS12 isoforms (UniProt P25398 isoforms). The record notes N-acetylalanine at residue 2 and N6-succinyllysine at residue 129; without a mapped antibody epitope, their effect on binding is unknown (UniProt P25398 modified residues; caption A01040-3). When staining shifts compartment or disappears, review retrieval, tissue preservation, and antibody controls before invoking epitope masking (standard IHC practice). A membrane-facing epitope explanation is unsupported by the supplied topology (UniProt P25398 topology).
How can IF help assess an ambiguous RPS12 IHC pattern?
Use the separate IF/ICC workflow to compare RPS12 with a marker identifying the cell population under examination; bronchial basal cells are one IHC reference population (HPA tissue IHC: High in bronchial basal cells; standard IF practice). Select fluorophores after inspecting tissue autofluorescence, and include single-colour controls when assessing overlap (standard IF practice). Because RPS12 has no transmembrane segment and is reported mainly in the cytosol, permeabilise cells sufficiently for intracellular antibody access while preserving morphology (UniProt P25398 topology; HPA subcellular: cytosol supported; standard IF practice). Assess any nucleolar or punctate signal against compartment markers and controls, since nucleolar localisation and additional Golgi or vesicular localisation are reported (UniProt P25398 subcellular location; HPA subcellular). Do not treat an IF pattern alone as validation of the paraffin IHC signal (standard assay validation practice).
How do I distinguish RPS12 signal from diffuse DAB background?
First examine a section processed without primary antibody to identify staining from the detection system or tissue itself (standard IHC practice). Quench endogenous peroxidase before HRP/DAB development and check whether brown deposits follow tissue edges, damaged areas, or pigment rather than intact cells (standard IHC practice). The selected paraffin example used 10% goat serum block, 1:100 primary overnight at 4°C, and a peroxidase-linked secondary for 30 minutes at 37°C (caption A01040-3). If background persists, compare washes and antibody concentration systematically while holding EDTA pH 8.0 retrieval constant (datasheet A01040-3; standard IHC practice). Interpret diffuse cytoplasmic staining against cell-resolved controls, since general cytoplasmic expression is reported (HPA tissue IHC).
What is a defensible way to score RPS12 chromogenic staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; RPS12 tissue IHC is described as generally cytoplasmic (HPA tissue IHC: general cytoplasmic expression). For each comparable region, report the percentage of positive intact cells and a prespecified intensity score, or combine those measurements as an H-score (standard IHC scoring practice). Normalise cell counts to the number of evaluable cells, or report positive-cell density per mm² of evaluable tissue when area is the denominator (standard IHC quantification practice). Exclude folds, necrosis, and section edges using the same rules for every image (standard IHC quantification practice). Keep retrieval, DAB development, counterstaining, and image settings consistent across groups (standard IHC quantification practice).
When is an apparent RPS12-positive cell likely to be an artefact?
Give greatest weight to staining within intact cells that fits the broadly cytoplasmic tissue pattern; nucleolar signal is also plausible for RPS12 (HPA tissue IHC: general cytoplasmic expression; UniProt P25398 subcellular location). Check cell identity: HPA reports High staining in bronchial basal cells but RPS12 is not detected in adipocytes, so the same brown deposit has different contextual support (HPA tissue IHC: bronchus basal cells High; adipose tissue adipocytes Not detected). Treat edge staining, necrotic deposits, and signal reproduced without primary antibody as artefact candidates (standard IHC practice). Check endogenous peroxidase when DAB appears in an unexpected compartment or cell population (standard IHC practice). HPA rates tissue staining Approved with medium consistency against RNA expression, so corroborate surprising patterns before making a biological claim (HPA tissue IHC: reliability).
Boster reagents
Best RPS12 / Small ribosomal subunit protein eS12 IHC Antibodies
The catalog antibody has real IHC images from human paraffin sections of thyroid, liver, bladder, and clear cell renal cancers (catalog: A01040-3 IHC captions). No IF figure is supplied (catalog: if_image_alts).
Real IHC data
Anti-RPS12 Antibody ®
Cat # A01040-3
A01040-3 is listed for IHC and Human, Mouse, and Rat reactivity (catalog: A01040-3 applications/reactivity). Its images document human paraffin-section IHC in thyroid, liver, bladder, and clear cell renal cancers (catalog: A01040-3 IHC captions).
Which to pick: Choose A01040-3 for tissue IHC: its own captions document paraffin sections, EDTA retrieval at pH 8.0, and 1:100 primary antibody incubation; the fixative is unreported (catalog: A01040-3 IHC captions). For IF/ICC, A01040-3 lists a 1:50–400 dilution range, but supplies no IF image (catalog: A01040-3 dilution_raw/if_image_alts). For cross-species studies, A01040-3 lists Human, Mouse, and Rat reactivity, while its IHC images show human tissue only (catalog: A01040-3 reactivity/IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P25398 (RS12_HUMAN, Small ribosomal subunit protein eS12).
- Human Protein Atlas. RPS12 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. RPS12 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the Golgi apparatus and vesicles..
- Human Protein Atlas. RPS12 antibody validation summary (2 antibodies).
- Proteomics of the temporal cortex in semantic dementia reveals brain-region specific molecular pathology and regulation of the TDP-43-ANXA11 interactome. Acta neuropathologica communications 2025 — PMC12291262.
- PubMed PMID:1861993 — UniProt-cited evidence.
- PubMed PMID:11875025 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RPS12 (Small ribosomal subunit protein eS12). Expected localisation General cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01040-3). Validated IHC antibody: A01040-3 at 1:50 recommended; image 1:100 (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Basal cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human thyroid cancer tissue; fixative not specified (datasheet A01040-3); 2. Fixation: Image fixative and duration unreported (datasheet A01040-3); 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 A01040-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01040-3); 8. Primary antibody: Rabbit anti-RPS12, 1:50 recommended; image 1:100 (datasheet A01040-3); 9. Primary incubation: Overnight at 4 °C (datasheet A01040-3). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining has medium consistency with RNA expression (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain spans residues 2–132 (UniProt); Regulation: Low tissue specificity; no regulator reported (HPA tissue IHC; UniProt). Evidence: UniProt P25398, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the Golgi…, Datasheet A01040-3, PMC12291262. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RPS12/immunohistochemistry-ihc-guide.