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RPL11 / Large ribosomal subunit protein uL5 · IHC design guide
Design Immunohistochemistry for RPL11
Plan paraffin-section IHC for RPL11 using the catalog antibody at 2–5 μg/ml (datasheet A02901-1). Assess cytoplasmic staining against tissue controls, while accounting for low consistency between antibody staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RPL11 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 in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02901-1) | |
| Positive control | Adrenal gland+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 conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Low staining–RNA consistency limits tissue predictions (HPA tissue IHC) | |
| Regulation | Expression regulator not annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended RPL11 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet A02901-1). Published IHC methods provide clinical-section and tissue-section examples (PMC12394147; PMC5693310).
| Sample | Paraffin-embedded human placenta tissue; fixative not specified (datasheet A02901-1) |
| Fixation | Image fixative and duration unreported (datasheet A02901-1); 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 A02901-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02901-1) |
| Primary antibody | Rabbit anti-RPL11, 2-5 μg/ml (datasheet A02901-1) |
| Primary incubation | Overnight at 4 °C (datasheet A02901-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02901-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RPL11-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet A02901-1); the published citrate pH 6.0 method used a different antibody (PMC5693310).
Section 2
What Is the Expected RPL11 Staining Pattern?
RPL11 is annotated in the nucleolus and cytoplasm, with no transmembrane segment (UniProt P62913). In tissue IHC, expect mainly cytoplasmic staining in many cell types; HPA reports medium staining in appendix glandular cells and bone marrow hematopoietic cells (HPA tissue IHC). Treat intensity as context dependent: the tissue IHC entry is Approved but reports low consistency between antibody staining and RNA expression (HPA tissue IHC).
| Cytoplasmic stain in appendix glandular cells or bone marrow hematopoietic cells. | This matches the reported medium signal in those cell types and the broader cytoplasmic tissue profile (HPA tissue IHC). Judge the named cells, rather than requiring uniform staining across the section (HPA tissue IHC). |
| Nucleolar or nuclear signal accompanies cytoplasmic stain. | Nucleolar localization is compatible with the UniProt annotation, but HPA tissue IHC chiefly describes cytoplasmic expression (UniProt P62913; HPA tissue IHC). Record each compartment separately; nuclear signal alone is insufficient to reproduce the reported tissue pattern (HPA tissue IHC). |
| Stain is confined to cell surfaces, lumens, or extracellular material. | These are outside the annotated nucleolar and cytoplasmic locations, and RPL11 has no transmembrane segment (UniProt P62913). Consider nonspecific deposition or detection artefact; inspect morphology and controls before scoring it as RPL11 (general IHC practice). |
| Strong signal appears in cardiomyocytes or adipocytes. | HPA lists those specific cell types as not detected; it does not label every cell in heart or adipose tissue negative (HPA tissue IHC). Confirm cell identity, then investigate cross-reactivity or endogenous detection activity (general IHC practice). |
| The entire slide is hazy, or appendix glandular cells show no signal. | Diffuse haze makes cell-level scoring unreliable (general IHC practice). Appendix glandular cells are reported at medium intensity, so a blank result there merits a technical check; HPA's low antibody–RNA consistency still limits any single negative inference (HPA tissue IHC). |
💡Expected RPL11 appearanceCall a result positive when identifiable cells show predominantly cytoplasmic signal, including medium staining in appendix glandular or bone marrow hematopoietic cells (HPA tissue IHC); isolated surface, luminal, or extracellular colour warrants artefact review (UniProt P62913; general IHC practice).
| Tissue and cell choice | HPA reports medium staining in several named populations, low staining in esophageal squamous cells and hepatocytes, and no detection in adipocytes or cardiomyocytes (HPA tissue IHC). Compare the same cell type across sections; a low-signal population is a weak positive control (general IHC practice). |
| Antibody evidence | The listed rabbit polyclonal HPA002734 is Approved for IHC; Enhanced validation is not reported, and antibody staining has low consistency with RNA expression (HPA tissue IHC; HPA antibodies). Interpret unexpected intensity with tissue morphology and controls rather than treating it as independently confirmed expression (general IHC practice). |
| Subcellular interpretation | UniProt places RPL11 in the nucleolus and cytoplasm and describes its role in the large ribosomal subunit and 5S RNP (UniProt P62913). HPA tissue IHC reports chiefly cytoplasmic staining, so record a nuclear component without requiring it for an IHC-positive call (HPA tissue IHC). |
| IF/ICC Q&A: Is an image-based RPL11 pattern established here? | No: the supplied HPA subcellular entry gives no main location or ICC-IF image-bearing cell line (HPA subcellular). UniProt's nucleolar and cytoplasmic annotation provides a biological expectation, but it is not an image-validated IF pattern in this record (UniProt P62913; HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in appendix glandular cells. | This conflicts with HPA's medium cell-level observation, although its IHC entry has low antibody–RNA consistency (HPA tissue IHC). A failed staining or detection run is also possible (general IHC practice). | Check section integrity, reagent controls, primary-antibody application, and chromogenic detection; review retrieval against the antibody's validated IHC conditions (general IHC practice). Do not infer RPL11 loss from one blank section. |
| Only hepatocytes or esophageal squamous cells look negative. | Both are listed as low, rather than medium, in the supplied tissue profile (HPA tissue IHC). A weak result there has limited value as a run-failure test (general IHC practice). | Assess a reported medium population, such as appendix glandular cells, on a comparable run before changing interpretation (HPA tissue IHC; general IHC practice). |
| Cardiomyocytes or adipocytes stain strongly. | HPA records no detection in those named cells (HPA tissue IHC). Cell identification error, cross-reactivity, or endogenous chromogenic activity may explain unexpected colour (general IHC practice). | Recheck morphology and a no-primary control; assess endogenous enzyme blocking if the detection chemistry requires it (general IHC practice). Report the affected cell type and compartment. |
| Brown colour covers tissue, edges, and empty spaces. | A pattern without cell boundaries is consistent with diffuse background or reagent deposition (general IHC practice); it cannot be matched to HPA's cytoplasmic cell-level profile (HPA tissue IHC). | Compare a no-primary control, inspect washing and blocking, and reassess the primary dilution using validated IHC conditions (general IHC practice). Score only resolved cellular signal. |
| Only membrane-like or extracellular staining remains. | That distribution does not fit the annotated nucleolar and cytoplasmic locations or absence of a transmembrane segment (UniProt P62913). | Check counterstained morphology and controls, then reassess detection background and antibody specificity before calling a positive result (general IHC practice). |
| Nuclear or nucleolar stain appears without clear cytoplasmic stain. | Nucleolar location is annotated, while the HPA tissue IHC profile emphasizes cytoplasmic staining (UniProt P62913; HPA tissue IHC). The two observations alone do not establish an IHC error. | Record the compartment, verify cell-level staining against controls, and compare a reported medium tissue population; avoid claiming an image-validated IF pattern from the HPA subcellular entry (HPA tissue IHC; HPA subcellular; general IHC practice). |
Experiment evidence
IHC & IF images showing RPL11 staining
A02901-1 · IHCIHC analysis of RPL11 using anti-RPL11 antibody (A02901-1). RPL11 was detected in a paraffin-embedded section of human placenta tissue. Heat
A02901-1 · IHCIHC analysis of RPL11 using anti-RPL11 antibody (A02901-1). RPL11 was detected in a paraffin-embedded section of mouse brain tissue. Heat me
A02901-1 · IFIF analysis of RPL11 using anti-RPL11 antibody (A02901-1). RPL11 was detected in an immunocytochemical section of MCF-7 cells. Enzyme antige
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000142676-RPL11/tissue/cerebral+cortex#imid_1141832
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000142676-RPL11/tissue/colon#imid_1141750
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000142676-RPL11/tissue/liver#imid_1141751
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000142676-RPL11/tissue/pancreas#imid_1141702
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RPL11 IHC & IF
🧪Run breast first: glandular cells should stain at the HPA Medium level (HPA: Breast glandular cells, Medium). Run adipose tissue as the negative comparator (HPA: Adipose tissue adipocytes, Not detected); adipocytes within the breast section, if present, should show little or no signal, although HPA has not validated them as an internal negative on that slide (HPA: adipocytes, Not detected in adipose tissue).
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for RPL11; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary and secondary-only controls, plus concentration-matched rabbit IgG isotype control for the rabbit primary (standard IHC controls; caption: rabbit anti-RPL11 antibody). For biological specificity, use matched RPL11-knockout material if viable, or peptide competition if the immunizing peptide is verified; quench endogenous peroxidase before HRP/DAB detection (standard IHC controls; caption: HRP/DAB detection).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A02901-1 paraffin-section caption does not state a fixative (caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 was used for the pictured IHC result, but whether retrieval is required is unreported; frozen-section performance and any ease advantage for IF/ICC are also unreported (caption: heat retrieval in EDTA; HPA subcellular: no ICC-IF images). In breast sections, score staining in cells rather than empty adipocyte vacuoles (standard paraffin IHC practice).
HPA tissue IHC evidence for RPL11
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | 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 → |
| Kidney | Cells in glomeruli | Not detected | Protein (IHC) | HPA → |
| Ovary | Follicle cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RPL11 IHC Tips
Use the documented paraffin section workflow as the starting point, then assess staining by compartment, cell type and tissue quality.
What should I change when RPL11 staining is weak after antigen retrieval?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A02901-1). The documented stain used 2 μg/ml primary antibody overnight at 4°C, so check those conditions before changing retrieval (datasheet A02901-1). If staining remains weak, compare a modest change in heating time on adjacent sections while keeping buffer, antibody concentration and detection constant (standard IHC practice). Include a section processed without primary antibody to distinguish weak target staining from detection background (standard IHC practice). Record whether any improvement occurs in nucleoli, nuclei or cytoplasm, the reported RPL11 compartments (UniProt P62913).
How can I troubleshoot suspected fixation effects on RPL11 IHC?
The selected image documents a paraffin section but does not state its fixative; RPL11-specific fixation sensitivity is therefore unknown (datasheet A02901-1). Record the fixative and fixation duration for each specimen, then compare sections processed with the same EDTA pH 8.0 retrieval and detection conditions (datasheet A02901-1; standard IHC practice). Check morphology alongside staining, because damaged tissue can make compartment assignment unreliable (standard IHC practice). Keep the documented 2 μg/ml overnight primary incubation constant during that comparison (datasheet A02901-1). Neither HPA staining patterns nor RPL11 topology and modifications establish a target-specific fixation effect (HPA tissue IHC; UniProt P62913).
Should RPL11 stain nucleoli, nuclei or cytoplasm in paraffin sections?
RPL11 is reported in the nucleolus, nucleus and cytoplasm, so assess each compartment separately rather than requiring one pattern in every cell (UniProt P62913). HPA describes cytoplasmic expression in most tissues, but its tissue IHC has low consistency with RNA expression data (HPA tissue IHC). Compare cells within intact regions at the same magnification and note whether a nuclear signal is discrete or diffuse (standard IHC practice). Use the no-primary control to check whether apparent nucleolar dots persist without the RPL11 antibody (standard IHC practice). Diffuse staining across damaged areas should prompt a morphology and background review before biological interpretation (standard IHC practice).
Could isoforms or epitope masking explain different RPL11 staining patterns?
RPL11 has 2 annotated isoforms, but the supplied evidence does not locate the catalog antibody’s epitope or establish isoform recognition (UniProt P62913; datasheet A02901-1). The annotated protein chain spans residues 2–178, with reported modifications including threonines 44 and 47 and lysine 85 (UniProt P62913). Those annotations alone cannot show that a modification blocks this antibody in a paraffin section (UniProt P62913). Compare retrieval conditions on adjacent sections while holding the documented antibody incubation constant (datasheet A02901-1; standard IHC practice). Interpret a changed pattern as an assay result until epitope mapping or independent antibody evidence supports an isoform explanation (standard IHC practice).
How should I investigate RPL11 localisation with multiplex IF after IHC?
Treat IF as a separate assay: the supplied HPA subcellular record lists no ICC/IF images, while the catalog example documents chromogenic tissue IHC (HPA subcellular; datasheet A02901-1). In bronchus, multiplex RPL11 with a validated respiratory epithelial cell marker to identify the cell population reported with medium staining by HPA (HPA tissue IHC; standard IF practice). Choose a fluorophore away from the specimen’s strongest autofluorescence and inspect single-channel and no-primary controls before interpreting overlap (standard IF practice). Permeabilise for access to intracellular nucleolar, nuclear and cytoplasmic RPL11; the antibody’s precise epitope remains unspecified (UniProt P62913; standard IF practice). Do not transfer the paraffin section retrieval conditions into an IF protocol without separate optimisation (datasheet A02901-1; standard IF practice).
What should I check when DAB obscures RPL11 staining?
Check a no-primary section and an appropriate peroxidase-block control before attributing widespread DAB deposition to RPL11 (standard IHC practice). The documented workflow used 10% goat serum blocking, a peroxidase-conjugated secondary and DAB development (datasheet A02901-1). If background persists, compare shorter chromogen development and more stringent washes while retaining the same retrieval and primary incubation (standard IHC practice). Inspect tissue edges, folds and damaged regions separately because staining there can distort the apparent distribution (standard IHC practice). HPA reports cytoplasmic expression in most tissues, so cytoplasmic colour alone does not establish specificity (HPA tissue IHC).
How should I score RPL11 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; RPL11 is reported in nucleoli, nuclei and cytoplasm (UniProt P62913). For chromogenic IHC, record the percentage of positive cells and an intensity-weighted H-score separately by compartment, using consistent thresholds across sections (standard IHC practice). Normalise positive counts to the number of evaluable cells of the same type, or report positive-cell density per mm² of viable tissue (standard IHC practice). Exclude folds, tissue edges and necrotic areas using prespecified image criteria (standard IHC practice). Keep retrieval, 2 μg/ml primary incubation and DAB development comparable between slides, and report HPA’s low RNA–staining consistency as a limit on interpretation (datasheet A02901-1; HPA tissue IHC).
How can I distinguish credible RPL11 signal from IHC artefact?
A credible result should align with intact cells and a reported RPL11 compartment: nucleolus, nucleus or cytoplasm (UniProt P62913; standard IHC practice). HPA reports medium staining in bronchial respiratory epithelial cells and no detection in cardiomyocytes, but also reports low consistency between staining and RNA expression (HPA tissue IHC). Treat those cell observations as context, not absolute positive or negative controls (HPA tissue IHC). Review isolated edge staining, necrosis and colour in no-primary or peroxidase-block controls before assigning DAB to RPL11 (standard IHC practice). If a signal appears only in an unexpected compartment or damaged cell population, repeat it on an adjacent section under the documented retrieval conditions (datasheet A02901-1; standard IHC practice).
Boster reagents
Best RPL11 / Large ribosomal subunit protein uL5 IHC Antibodies
A02901-1 has IHC images of paraffin sections from human placenta and mouse and rat brain, plus an IF/ICC image of MCF-7 cells (catalog image captions).
Real IHC data
Anti-RPL11 Antibody ®
Cat # A02901-1
A02901-1 is listed for IHC and shows staining in paraffin sections of human placenta and mouse and rat brain (catalog applications; A02901-1 IHC captions). The same SKU is listed for IF/ICC and shows staining in MCF-7 cells (catalog applications; A02901-1 IF caption).
Which to pick: Choose A02901-1 for paraffin-section IHC: its captions document EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml; the fixative is unreported (A02901-1 IHC captions). For IF/ICC, A02901-1 has an MCF-7 cell image at 5 μg/ml (A02901-1 IF caption). It is also the listed cross-species choice for human, mouse and rat IHC; clonality is unreported (catalog reactivity; A02901-1 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P62913 (RL11_HUMAN, Large ribosomal subunit protein uL5).
- Human Protein Atlas. RPL11 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. RPL11 subcellular location (ICC-IF): Highest expression in HTCEpi: 2976.3 nTPM.
- Human Protein Atlas. RPL11 antibody validation summary (1 antibodies).
- Silencing RPL11 attenuates acute kidney injury by suppressing tubular apoptosis and macrophage-driven inflammation. Frontiers in immunology 2025 — PMC12394147.
- Loss of RPS27a expression regulates the cell cycle, apoptosis, and proliferation via the RPL11-MDM2-p53 pathway in lung adenocarcinoma cells. Journal of experimental & clinical cancer research : CR 2022 — PMC8785590.
- Ribosomal protein RPL11 haploinsufficiency causes anemia in mice via activation of the RP-MDM2-p53 pathway. The Journal of biological chemistry 2023 — PMC9793318.
- Disruption of the RP-MDM2-p53 pathway accelerates APC loss-induced colorectal tumorigenesis. Oncogene 2017 — PMC5693310.
- PubMed PMID:7748210 — UniProt-cited evidence.
- PubMed PMID:12815950 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RPL11 (Large ribosomal subunit protein uL5). Expected localisation Cytoplasmic in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02901-1). Validated IHC antibody: A02901-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human placenta tissue; fixative not specified (datasheet A02901-1); 2. Fixation: Image fixative and duration unreported (datasheet A02901-1); 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 A02901-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02901-1); 8. Primary antibody: Rabbit anti-RPL11, 2-5 μg/ml (datasheet A02901-1); 9. Primary incubation: Overnight at 4 °C (datasheet A02901-1). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Low staining–RNA consistency limits tissue predictions (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unknown (UniProt); Regulation: Expression regulator not annotated (UniProt). Evidence: UniProt P62913, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in HTCEpi: 2976.3 nTPM, Datasheet A02901-1, PMC12394147, PMC5693310. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RPL11/immunohistochemistry-ihc-guide.