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RPS26 / Small ribosomal subunit protein eS26 · IHC design guide
Design Immunohistochemistry for RPS26
Plan chromogenic RPS26 IHC around cytoplasmic staining in several tissues (HPA tissue IHC). Use the catalog antibody’s paraffin-section protocol (datasheet A06454), and interpret tissue differences cautiously because HPA rates its staining reliability as uncertain (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RPS26 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 glandular cells and cholangiocytes (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06454) | |
| Positive control | Gallbladder+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); optimize as needed. | |
| Caveat | Tissue staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Low tissue specificity; no regulator annotated (HPA tissue IHC; UniProt) | |
| Isoform / epitope | 0 isoforms annotated; chain spans residues 2–115 (UniProt) |
Section 1
Recommended RPS26 IHC & IF Protocols
The catalog antibody IHC-P protocol (datasheet A06454) is paired with one published mouse embryo IHC protocol (PMC5333091).
| Sample | Paraffin-embedded human kidney tissue; fixative not specified (datasheet A06454) |
| Fixation | Image fixative and duration unreported (datasheet A06454); 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 A06454); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06454) |
| Primary antibody | Rabbit anti-RPS26, 1:50 recommended; image 2 μg/ml (datasheet A06454) |
| Primary incubation | Overnight at 4 °C (datasheet A06454) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06454) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RPS26-positive staining in glandular cells of gallbladder (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 A06454). The published protocol specifies high-pressure retrieval without naming a buffer (PMC5333091).
Section 2
What Is the Expected RPS26 Staining Pattern?
RPS26 is a 40S ribosomal protein detected on cytosolic polysomes and in ribosomes associated with rough endoplasmic reticulum; it has no transmembrane segment (UniProt P62854). In paraffin-section IHC, expect cytoplasmic staining, including in gallbladder glandular cells, liver cholangiocytes and pancreatic exocrine glandular cells (HPA: High in each). Treat intensity comparisons cautiously: HPA rates tissue IHC reliability Uncertain because staining and RNA expression have low consistency (HPA: tissue IHC).
| Cytoplasmic staining in gallbladder glandular cells, cholangiocytes or pancreatic exocrine glandular cells. | This matches the reported High IHC staining in those cell types (HPA: tissue IHC). Cytoplasmic localisation also fits RPS26 on cytosolic polysomes and rough-ER-associated ribosomes (UniProt P62854). Judge the pattern by cell type as well as intensity because the tissue IHC assessment is Uncertain (HPA: tissue IHC). |
| Strong chromogenic signal confined to nuclei, with little cytoplasmic staining. | A nuclear-only IHC pattern does not match the reported cytoplasmic tissue profile (HPA: tissue IHC). Check morphology, counterstain and detection controls before interpreting it as RPS26 (general IHC practice). Nucleolar signal is an additional ICC-IF location, so that IF finding alone does not validate nuclear-only chromogenic IHC (HPA: subcellular). |
| Strong staining in adipocytes, cardiomyocytes or cells in kidney glomeruli. | Those cell types were Not detected in the supplied tissue IHC profile (HPA: tissue IHC). Unexpected staining could reflect cross-reactivity or endogenous detection activity (general IHC practice); inspect controls and tissue morphology. These HPA observations are comparison points, not proof that every specimen of those tissues lacks RPS26 (HPA: IHC reliability Uncertain). |
| Uniform colour over cells, stroma and empty spaces, without a recognisable cytoplasmic pattern. | Diffuse staining obscures the cell-specific cytoplasmic profile reported for RPS26 (HPA: tissue IHC). Assess background with a primary-omission control and review blocking, washing and detection chemistry (general IHC practice). Background intensity alone cannot be scored as target expression when cellular boundaries and localisation are unclear (general IHC practice). |
| No stain in gallbladder glandular cells, cholangiocytes or pancreatic exocrine glandular cells. | Absence in these reported High groups warrants a technical check before calling the sample negative (HPA: tissue IHC). Verify that the cells are present and assess the primary antibody and detection controls (general IHC practice). The HPA tissue pattern is Uncertain, so a single negative section cannot resolve expression on its own (HPA: tissue IHC). |
💡Expected RPS26 appearanceA credible positive is cell-associated cytoplasmic chromogen, potentially High in gallbladder glandular cells, cholangiocytes or pancreatic exocrine glandular cells (HPA: tissue IHC); uniform background or nuclear-only colour is suspect (general IHC practice; HPA: tissue IHC).
| Ribosomal location | RPS26 is part of the 40S subunit and occurs on cytosolic polysomes and rough-ER-associated ribosomes (UniProt P62854). Interpret an intracellular cytoplasmic pattern in light of this distribution; the record reports no transmembrane segment (UniProt P62854). |
| Strength of tissue evidence | HPA calls the IHC tissue profile Uncertain because antibody staining has low consistency with RNA expression; RNA shows low tissue specificity (HPA: tissue IHC). Use listed High and Not detected groups as observed comparators, rather than absolute expression rules (HPA: tissue IHC). |
| IF/ICC: where is signal expected? | Mainly endoplasmic reticulum and cytosol, with additional nucleolar localisation (HPA: subcellular ICC-IF). This is an IF localisation reference, not an IHC protocol: HPA043961 is ICC Approved but IHC Uncertain, while HPA055803 is ICC Approved with no IHC status listed (HPA: antibodies). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected cytoplasmic stain is absent in a reported High group (HPA: tissue IHC). | Missing target cells or an unsuccessful staining run are possible (general IHC practice). | Confirm cell identity and section quality, then review primary-antibody and detection controls before interpreting absence (general IHC practice; HPA: IHC reliability Uncertain). |
| Colour appears across the whole section, including spaces without cells. | Nonspecific background or detection chemistry may obscure localisation (general IHC practice). | Compare a primary-omission control and review blocking and washing; score only interpretable, cell-associated cytoplasmic signal (general IHC practice; HPA: cytoplasmic tissue profile). |
| Nuclear-only chromogen dominates the section. | The pattern conflicts with HPA's cytoplasmic tissue profile; its cause is unresolved (HPA: tissue IHC). | Check counterstain and detection controls, then reassess whether the stain follows cytoplasm in identifiable cells (general IHC practice; HPA: tissue IHC). |
| A reported Not detected cell group stains strongly (HPA: tissue IHC). | Cross-reactivity or endogenous detection activity is possible (general IHC practice). | Check primary-omission and detection controls and compare with a reported High cell group; avoid treating one discordant specimen as definitive (general IHC practice; HPA: IHC reliability Uncertain). |
| Intensity differs between two listed tissue groups. | The supplied profile spans High, Medium, Low and Not detected observations, and its IHC reliability is Uncertain (HPA: tissue IHC). | Record cell type, compartment and intensity separately; compare equivalent cells under the same scoring criteria before drawing an expression conclusion (general IHC practice; HPA: tissue IHC). |
| An IF image shows nucleolar signal alongside cytosol or ER signal. | Nucleoli are an additional ICC-IF location; ER and cytosol are the main locations (HPA: subcellular ICC-IF). | Interpret the IF compartment pattern using the separate IF/ICC guide; do not use it to override the cytoplasmic paraffin IHC profile (HPA: subcellular ICC-IF; HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing RPS26 staining
A06454 · IHCIHC analysis of RPS26 using anti-RPS26 antibody (A06454). RPS26 was detected in a paraffin-embedded section of human kidney tissue. Heat med
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000197728-RPS26/tissue/cerebral+cortex#imid_12170345
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000197728-RPS26/tissue/colon#imid_12170271
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000197728-RPS26/tissue/liver#imid_12170288
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000197728-RPS26/tissue/kidney#imid_12170273
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043961; green = RPS26, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043961; green = RPS26, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RPS26 IHC & IF
🧪Run gallbladder first: glandular cells should stain (HPA: High in gallbladder glandular cells). Use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipocytes); on the gallbladder slide, treat non-glandular cells as internal background comparators only, because their RPS26 status is not supplied (HPA: gallbladder glandular cells High).
Positive control tissue: Gallbladder (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RPS26 in A-431, U-251MG, U2OS, with annotated localisation: Endoplasmic reticulum (enhanced), Cytosol (enhanced) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls (selected A06454 caption: rabbit primary antibody; standard IHC practice), plus a matched RPS26 knockout specimen as a biological specificity control (standard IHC practice). Block endogenous peroxidase before HRP/DAB detection and assess any gallbladder pigment separately from chromogen signal (selected A06454 caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A06454 paraffin-section caption does not state the fixative (selected A06454 caption). Heat retrieval in EDTA at pH 8.0 is documented, but whether staining depends on that retrieval condition is unreported (selected A06454 caption). Frozen sections have no supplied protocol, while ICC-IF images show endoplasmic reticulum and cytosol localization with additional nucleolar signal; neither establishes an easier alternative to paraffin IHC (HPA: subcellular ICC-IF; selected A06454 caption).
HPA tissue IHC evidence for RPS26
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low 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 |
|---|---|---|---|---|
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | High | Protein (IHC) | HPA → |
| Pancreas | Exocrine glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RPS26 IHC Tips
Troubleshoot RPS26 staining in paraffin sections by checking retrieval, cytoplasmic localisation, assay controls, and cell-specific scoring (datasheet A06454; UniProt P62854; HPA tissue IHC).
How should I optimise retrieval when RPS26 staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for the catalog antibody (datasheet A06454). The kidney section shown for A06454 was paraffin embedded, but its caption gives no heating time or fixative, so establish a local retrieval time series, such as 10–20 minutes, while keeping section thickness and detection constant (datasheet A06454; standard IHC practice). If staining remains weak, compare a citrate buffer at pH 6.0 as a fallback on matched sections and inspect morphology alongside signal (standard IHC practice). Include a no-primary control and score cytoplasmic staining in the same cell population before selecting a condition (UniProt P62854 subcellular location; standard IHC practice).
Could fixation explain weak or uneven RPS26 staining?
Target-specific RPS26 fixation sensitivity is unknown: the A06454 kidney caption identifies a paraffin section but does not state its fixative (datasheet A06454). Record fixative, time before fixation, and fixation duration for every specimen, then compare similarly processed sections before attributing a difference to biology (standard IHC practice). For a new series, use a consistent local fixation procedure and pilot retrieval on matched sections; do not infer an RPS26-specific fixation response from its cytoplasmic location or phosphorylation (UniProt P62854; standard IHC practice). Check section adhesion, preservation, and regional staining before increasing primary antibody concentration beyond the caption's 2 µg/ml (datasheet A06454; standard IHC practice).
Where should convincing RPS26 signal appear in a tissue section?
Expect predominantly cytoplasmic signal because RPS26 is a 40S ribosomal component detected on cytosolic polysomes and associated with rough endoplasmic reticulum (UniProt P62854). HPA tissue IHC describes cytoplasmic expression in several tissues, although its antibody staining has uncertain reliability (HPA tissue IHC). A fine cytoplasmic pattern can fit the target, while isolated sharp nuclear staining warrants checking counterstain, background, and antibody specificity before interpretation (UniProt P62854; standard IHC practice). HPA cell imaging additionally reports nucleolar localisation, so a nucleolar component should be evaluated with appropriate controls rather than dismissed solely by compartment (HPA subcellular; standard IHC practice).
Could epitope accessibility or RPS26 processing change my result?
RPS26 has no annotated isoforms, signal peptide, propeptide, or transmembrane segment; its annotated mature chain spans residues 2–115 (UniProt P62854). A phosphoserine is recorded at residue 54, but the supplied evidence does not map the A06454 epitope or establish phosphorylation-dependent staining (UniProt P62854; datasheet A06454). Compare matched retrieval conditions and antibody concentrations before assigning a signal difference to epitope masking, and keep the same detection settings across that comparison (standard IHC practice). If a compartment or tissue pattern is unexpected, verify it with an independent antibody recognising a documented different epitope when available (standard IHC practice).
How can I assess RPS26 by IF alongside a cell identity marker?
For the separate IF/ICC workflow, multiplex RPS26 with a validated marker of the cell type being evaluated, such as cholangiocytes where tissue IHC reports high staining (HPA tissue IHC; standard IF practice). Select spectrally separated fluorophores and place the weaker signal in a channel with lower tissue autofluorescence; include single-stain and no-primary controls (standard IF practice). Permeabilise sufficiently to expose cytosolic and rough-ER-associated ribosomes, while titrating detergent because the A06454 epitope is unmapped and RPS26 has no transmembrane segment (UniProt P62854; datasheet A06454; standard IF practice). HPA cell imaging places RPS26 mainly in ER and cytosol, with additional nucleolar signal; confirm any such pattern under the chosen fixation conditions (HPA subcellular; standard IF practice).
What should I check when DAB appears throughout the section?
The A06454 kidney example used 10% goat serum, 2 µg/ml primary antibody overnight at 4°C, and an HRP/DAB detection system (datasheet A06454). Check a no-primary section first, then assess whether background follows endogenous peroxidase activity, secondary antibody binding, or excessive DAB development (standard IHC practice). Apply a peroxidase block and optimise blocking, washes, primary concentration, and chromogen exposure on matched sections; these are general IHC controls, not demonstrated RPS26-specific remedies (standard IHC practice). Compare background with the expected cytoplasmic distribution and avoid scoring diffuse deposits or staining confined to damaged section edges as RPS26 (UniProt P62854; standard IHC practice).
How should I score RPS26 across specimens with different cellular composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic compartment before scoring, because RPS26 is a ribosomal protein with a predominantly cytoplasmic distribution (UniProt P62854; HPA tissue IHC). For chromogenic sections, report the percentage of positive target cells and an H-score using intensity grades 0–3, or positive-cell density per mm² of viable tissue (standard IHC practice). Normalise counts to the number of evaluable cells in the same annotated compartment, and apply identical thresholds, exposure, and region-selection rules across sections (standard IHC practice). Report cell types separately: HPA records high staining in cholangiocytes but no detection in kidney glomerular cells, with overall uncertain IHC reliability (HPA tissue IHC).
How can I distinguish true RPS26 staining from an artefact?
A credible pattern includes cytoplasmic staining in morphologically intact cells, consistent with RPS26 on cytosolic and rough-ER-associated ribosomes (UniProt P62854). Cross-check cell identity: HPA reports high cholangiocyte staining but no detection in kidney glomerular cells, and rates its tissue IHC reliability uncertain (HPA tissue IHC). Flag staining limited to tissue edges, necrotic areas, or blood-rich regions, and compare no-primary and peroxidase-block controls before calling it specific DAB signal (standard IHC practice). Strong isolated nuclear staining or a uniform precipitate warrants reassessing localisation, retrieval, and detection rather than assigning a new RPS26 distribution from one section (UniProt P62854; standard IHC practice).
Boster reagents
Best RPS26 / Small ribosomal subunit protein eS26 IHC Antibodies
A06454 has real IHC data from a human paraffin-embedded kidney section (A06454 image caption). IF/ICC is listed as an application; human, mouse and rat are listed as reactive species (A06454 catalog).
Real IHC data
Anti-RPS26 Antibody
Cat # A06454
A06454 will render with its own IHC figure showing RPS26 staining in a human paraffin-embedded kidney section (A06454 image caption). The catalog lists IHC, IF and ICC applications and human, mouse and rat reactivity; it provides no IF image (A06454 catalog).
Which to pick: Choose A06454 for paraffin-section IHC: its figure documents heat retrieval in EDTA at pH 8.0, overnight primary incubation at 4°C and chromogenic detection in human kidney (A06454 image caption). A06454 is also the listed choice for IF/ICC and for evaluating mouse or rat samples because it is polyclonal and those applications and species appear in its catalog entry; the supplied figure demonstrates human IHC only (A06454 catalog; A06454 image caption). The IHC caption does not report the tissue fixative (A06454 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P62854 (RS26_HUMAN, Small ribosomal subunit protein eS26).
- Human Protein Atlas. RPS26 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. RPS26 subcellular location (ICC-IF): Mainly localized to the endoplasmic reticulum and cytosol. In addition localized to the nucleoli..
- Human Protein Atlas. RPS26 antibody validation summary (2 antibodies).
- Genome-wide analyses of non-syndromic cleft lip with palate identify 14 novel loci and genetic heterogeneity. Nature communications 2017 — PMC5333091.
- PubMed PMID:8464749 — UniProt-cited evidence.
- PubMed PMID:7945460 — UniProt-cited evidence.
- PubMed PMID:9602156 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RPS26 (Small ribosomal subunit protein eS26). Expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06454). Validated IHC antibody: A06454 at 1:50 recommended; image 2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Gallbladder (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human kidney tissue; fixative not specified (datasheet A06454); 2. Fixation: Image fixative and duration unreported (datasheet A06454); 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 A06454); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06454); 8. Primary antibody: Rabbit anti-RPS26, 1:50 recommended; image 2 μg/ml (datasheet A06454); 9. Primary incubation: Overnight at 4 °C (datasheet A06454). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); optimize as needed.; Caveat: Tissue staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 0 isoforms annotated; chain spans residues 2–115 (UniProt); Regulation: Low tissue specificity; no regulator annotated (HPA tissue IHC; UniProt). Evidence: UniProt P62854, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the endoplasmic reticulum and cytosol. In…, Datasheet A06454, PMC5333091. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RPS26/immunohistochemistry-ihc-guide.