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RPL36 / Large ribosomal subunit protein eL36 · IHC design guide
Design Immunohistochemistry for RPL36
Plan chromogenic RPL36 IHC around its general cytoplasmic tissue staining (HPA tissue IHC). Use strongly staining breast glandular cells and weakly staining adipocytes to help assess the staining range (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RPL36 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 | Broad cytoplasmic staining; glandular cells can stain strongly (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A08922-3) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Adipocytes and ovarian stroma show low staining (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | No isoforms annotated; mature chain spans residues 2–105 (UniProt) |
Section 1
Recommended RPL36 IHC & IF Protocols
The catalog antibody’s paraffin-section IHC protocol (datasheet: A08922-3) is accompanied by one published frozen-section RPL36 IHC protocol (PMC5742712).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A08922-3) |
| Fixation | Image fixative and duration unreported (datasheet A08922-3); 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 A08922-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08922-3) |
| Primary antibody | Rabbit anti-RPL36, 2-5 μg/ml (datasheet A08922-3) |
| Primary incubation | Overnight at 4 °C (datasheet A08922-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A08922-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RPL36-positive staining in endocrine cells of appendix (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteFor paraffin sections, start with heat-mediated EDTA retrieval at pH 8.0 (datasheet: A08922-3). The published frozen-section protocol used acetone fixation (PMC5742712).
Section 2
What Is the Expected RPL36 Staining Pattern?
RPL36 is a large ribosomal subunit protein detected on cytosolic polysomes, with no transmembrane segment (UniProt Q9Y3U8). In paraffin-section IHC, expect predominantly cytoplasmic staining across several cell types, including appendix endocrine cells, bone-marrow hematopoietic cells and breast glandular cells reported as High (HPA tissue IHC). HPA calls the tissue staining Supported, with medium consistency between antibody staining and RNA expression (HPA tissue IHC).
| Clear cytoplasmic staining in appendix endocrine cells, bone-marrow hematopoietic cells or breast glandular cells. | This agrees with the reported High staining in those cell populations and the general cytoplasmic profile (HPA tissue IHC). Interpret the compartment and the identified cell population together; a strong signal alone is less informative when surrounding tissue also stains. The cytosolic location is consistent with RPL36 on polysomes (UniProt Q9Y3U8). |
| Predominantly nuclear, membranous or extracellular staining, with little cytoplasmic signal. | This does not fit the reported cytoplasmic pattern (HPA tissue IHC) or cytosolic localization (UniProt Q9Y3U8). Check the corresponding no-primary control, tissue morphology and detection reagents before assigning the signal to RPL36 (general IHC practice). RPL36 has no annotated transmembrane segment (UniProt Q9Y3U8). |
| Signal is concentrated in adipocytes or ovarian stroma while expected High-staining cells appear weak. | HPA reports Low staining in adipocytes and ovarian stroma, versus High staining in the listed positive cell populations (HPA tissue IHC). This reversal warrants investigation for cross-reactivity, endogenous detection activity or cell-identification error (general IHC practice). Low does not mean absent, so neither population is a validated negative control (HPA tissue IHC). |
| Diffuse color covers cells and extracellular areas without a readable cytoplasmic pattern. | Treat the result as background until a no-primary control and the tissue morphology establish where signal originates (general IHC practice). Excess detection signal or inadequate blocking can obscure compartment boundaries (general IHC practice). Diffuse staining by itself cannot confirm the general cytoplasmic RPL36 profile reported by HPA (HPA tissue IHC). |
| No visible signal in a section containing an expected High-staining cell population. | First verify that the relevant cells are present and preserved; HPA reports High staining for the named populations, not for every cell in their tissues (HPA tissue IHC). Then review antibody application, retrieval, detection and control performance as general IHC variables (general IHC practice). A blank slide alone cannot establish RPL36 absence. |
💡Expected RPL36 appearanceCall a result consistent when staining is predominantly cytoplasmic in identified High-staining populations, such as bone-marrow hematopoietic cells (HPA tissue IHC); isolated nuclear, membranous or extracellular color is suspect given the cytosolic localization (UniProt Q9Y3U8).
| Cell population and tissue context | RPL36 has Low tissue RNA specificity, while HPA reports High staining in selected populations and Low staining in adipocytes and ovarian stroma (HPA tissue IHC). Compare like cell populations across sections; tissue-wide intensity averages can hide the expected pattern (general IHC practice). |
| Antibody evidence | HPA047153 is listed as Supported for IHC, and the tissue profile is Supported with medium staining–RNA consistency (HPA antibody record; HPA tissue IHC). These labels support pattern comparison but do not make every unexpected signal specific. Use controls when localization conflicts with the reported cytoplasmic profile (general IHC practice). |
| IF/ICC: where should RPL36 appear? | The expected location is the cytosol (HPA ICC-IF; UniProt Q9Y3U8). HPA lists ICC-IF images for HEK293, PC-3 and U2OS and marks HPA047153 Supported for ICC (HPA ICC-IF; HPA antibody record). This localization answers the IF/ICC question without implying that paraffin-section IHC settings transfer to IF. |
| Situation | Likely cause | Next action |
|---|---|---|
| A known High-staining population is blank. | The relevant cells may be absent from the section, or an IHC workflow step may have failed (HPA tissue IHC; general IHC practice). | Confirm cell identity and morphology, then check a positive control and the primary-antibody and detection steps (general IHC practice). Optimize retrieval empirically if needed; target-specific fixation sensitivity is unreported in the supplied evidence. |
| Signal is mostly nuclear or outlines cell membranes. | The distribution conflicts with cytosolic localization and the general cytoplasmic tissue profile (UniProt Q9Y3U8; HPA tissue IHC). | Inspect morphology and the no-primary control; reassess antibody and detection specificity before scoring those compartments as positive (general IHC practice). |
| Color appears where no primary antibody was applied. | The primary antibody cannot explain that control signal; endogenous detection activity or detection-reagent background is possible (general IHC practice). | Review blocking appropriate to the detection chemistry and the secondary or chromogen controls, then repeat with a clean no-primary control (general IHC practice). |
| Cytoplasmic boundaries disappear under widespread dark staining. | Overdeveloped chromogen or excessive assay signal can prevent compartment assessment (general IHC practice). | Reduce development or adjust assay conditions using matched controls, then score identifiable cells for a cytoplasmic pattern (general IHC practice; HPA tissue IHC). |
| Adipocytes or ovarian stroma look stronger than the selected positive cells. | That ranking differs from HPA's Low reports for those cells and High reports for selected positive populations (HPA tissue IHC). | Check cell identification, no-primary background and detection activity; compare matched sections before inferring cross-reactivity (general IHC practice). |
| Only weak staining is seen in a tissue outside HPA's listed High populations. | HPA reports general cytoplasmic expression and Low tissue RNA specificity, without a definitive negative tissue in this payload (HPA tissue IHC). | Score the identified cell population and compartment, and compare a listed High-staining population in the same run; avoid treating unlisted tissue as a negative control (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing RPL36 staining
A08922-3 · IHCIHC analysis of RPL36 using anti-RPL36 antibody (A08922-3). RPL36 was detected in a paraffin-embedded section of human breast cancer tissue.
A08922-3 · IHCIHC analysis of RPL36 using anti-RPL36 antibody (A08922-3). RPL36 was detected in a paraffin-embedded section of human cervical cancer tissu
A08922-3 · IFIF analysis of RPL36 using anti-RPL36 antibody (A08922-3). RPL36 was detected in a paraffin-embedded section of human colon cancer tissue. H
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000130255-RPL36/tissue/cerebral+cortex#imid_13160607
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000130255-RPL36/tissue/colon#imid_13160397
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000130255-RPL36/tissue/liver#imid_13160382
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000130255-RPL36/tissue/kidney#imid_13160391
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA047153; green = RPL36, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA047153; green = RPL36, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RPL36 IHC & IF
🧪Run breast first: glandular cells should show RPL36 staining (HPA: High in breast glandular cells; selected SKU caption: paraffin-section breast cancer IHC). HPA detects RPL36 in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and treat cells showing only counterstain without cytosolic DAB as a background reference, not validated internal negatives (HPA: no negative tissue rows; HPA: cytosolic localization).
Positive control tissue: Appendix (Endocrine cells, HPA High)
Negative control tissue: None in HPA: RPL36 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RPL36 in HEK293, PC-3, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a matched rabbit IgG isotype control with matching clonality if known, and RPL36 knockout material if available as a biological negative (selected SKU caption: rabbit primary; standard IHC controls). Quench endogenous peroxidase and check for residual DAB signal in the breast section (selected SKU caption: HRP/DAB detection; standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected SKU paraffin-section caption does not state a fixative (selected SKU caption). That caption uses heat retrieval in EDTA at pH 8.0, but it does not establish that retrieval is required; paraffin-section IHC is demonstrated, while the available IF images do not establish that frozen sections or IF are easier (selected SKU caption: EDTA retrieval and breast cancer IHC; HPA: ICC-IF images in HEK293, PC-3 and U2OS). Endogenous peroxidase can produce misleading DAB signal in the breast section, so interpret cytosolic staining against the no-primary control (selected SKU caption: HRP/DAB detection; standard chromogenic IHC practice).
HPA tissue IHC evidence for RPL36
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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 |
|---|---|---|---|---|
| Appendix | Endocrine cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: RPL36 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced RPL36 IHC Tips
Use cytoplasmic localisation, matched controls and consistent staining conditions to troubleshoot RPL36 chromogenic IHC in paraffin sections.
What retrieval should I try first if RPL36 staining is weak?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A08922-3). The selected tissue image used this retrieval before incubation with 2 μg/ml primary antibody overnight at 4°C, so reproduce those conditions when assessing weak signal (caption A08922-3). If staining remains weak, compare retrieval heating and cooling conditions on adjacent sections while keeping antibody concentration and detection constant (standard IHC practice). Include a tissue section with an expected cytoplasmic signal, such as breast glandular cells, and inspect morphology because excessive heating can damage sections (HPA: High in breast glandular cells; standard IHC practice).
Could fixation explain weak or uneven RPL36 staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative, and the supplied evidence does not establish how fixation changes RPL36 detection (caption A08922-3). Record the fixative, fixation duration and processing history for each specimen before attributing a weak result to antigen loss (standard IHC practice). Compare similarly processed sections with the same EDTA pH 8.0 retrieval and 2 μg/ml antibody incubation used in the selected image (datasheet A08922-3; caption A08922-3). Review section morphology and staining uniformity alongside the signal, since processing differences can complicate IHC comparisons (standard IHC practice).
Where should a convincing RPL36 signal appear in tissue sections?
Expect predominantly cytoplasmic staining: RPL36 is reported in the cytosol and on cytosolic polysomes, and HPA describes general cytoplasmic tissue expression (UniProt Q9Y3U8 subcellular location; HPA tissue IHC profile). It has no annotated transmembrane segment, so a crisp membrane-only pattern is inconsistent with the supplied localisation evidence (UniProt Q9Y3U8 topology). Compare stained cells with adjacent morphology and a hematoxylin counterstain to distinguish cytoplasm from overlapping nuclei or extracellular pigment (standard IHC practice). If staining appears exclusively nuclear or membranous, review background controls, retrieval and antibody specificity before assigning that pattern to RPL36 (UniProt Q9Y3U8 subcellular location; standard IHC practice).
How should I assess an unexpected staining pattern when the epitope is unspecified?
The supplied record annotates 0 isoforms and no protein domains, but it does not identify the catalog antibody's epitope (UniProt Q9Y3U8 record; caption A08922-3). RPL36 has an annotated acetyllysine at residue 62; the supplied evidence does not show whether that modification affects antibody binding (UniProt Q9Y3U8 modified residues). Keep retrieval at EDTA pH 8.0 while comparing serial sections, so a changed pattern is easier to interpret (datasheet A08922-3; standard IHC practice). If an independent antibody with a documented, distinct epitope is available, compare its cytoplasmic distribution and appropriate controls before inferring epitope masking (UniProt Q9Y3U8 subcellular location; standard IHC practice).
How can IF help check an ambiguous chromogenic RPL36 pattern?
Use IF as a separate localisation check, since HPA reports supported cytosolic localisation from ICC/IF images in HEK293, PC-3 and U2OS cells (HPA subcellular). For tissue multiplexing, pair RPL36 with a marker for the cell population under examination, such as glandular cells when assessing breast tissue, and confirm that signals occupy the same cells (HPA: High in breast glandular cells; standard IF practice). Choose spectrally separated fluorophores and check an unstained tissue section for autofluorescence before interpreting overlap (standard IF practice). Because RPL36 is cytosolic and has no transmembrane segment, use controlled permeabilisation to allow antibody access to the intracellular epitope, then assess background in matched controls (UniProt Q9Y3U8 subcellular location and topology; standard IF practice).
How can I reduce diffuse brown staining without losing RPL36 signal?
First distinguish diffuse cytoplasmic signal from background, since general cytoplasmic expression is reported for RPL36 in tissue (HPA tissue IHC profile). The selected image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and DAB detection; these provide documented starting conditions for this SKU (caption A08922-3). Include a no-primary control, check washes and apply an appropriate peroxidase block to assess nonspecific chromogen deposition (standard IHC practice). If background persists, compare antibody concentration and detection development on matched sections while checking that expected cytoplasmic staining remains visible (UniProt Q9Y3U8 subcellular location; standard IHC practice).
How should I score RPL36 staining across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic compartment before scoring, because RPL36 has general cytoplasmic tissue expression and low RNA tissue specificity (HPA tissue IHC profile; HPA RNA specificity). For chromogenic sections, record the percentage of positive cells and staining intensity, then calculate an H-score with a 0–300 range if intensity is graded 0–3 (standard IHC scoring practice). Alternatively, report positive-cell density per mm² within a defined tissue area, excluding folds, necrosis and section edges (standard IHC practice). Normalise comparisons to the same eligible cell population or tissue area, and keep retrieval, antibody incubation and image analysis settings consistent (standard IHC practice; datasheet A08922-3).
When should I question an apparent RPL36-positive result?
A credible result shows cytoplasmic staining in morphologically intact cells, consistent with the reported cytosolic location and general cytoplasmic tissue profile (UniProt Q9Y3U8 subcellular location; HPA tissue IHC profile). Question an exclusively membranous or nuclear pattern because it conflicts with the supplied localisation and topology evidence (UniProt Q9Y3U8 subcellular location and topology). Also inspect section edges, necrotic regions and no-primary controls for staining caused by processing or endogenous peroxidase activity in a DAB workflow (standard IHC practice). HPA rates tissue IHC reliability as Supported with medium staining-to-RNA consistency, so interpret intensity alongside cell identity, morphology and controls (HPA tissue IHC reliability; standard IHC practice).
Boster reagents
Best RPL36 / Large ribosomal subunit protein eL36 IHC Antibodies
A08922-3 has IHC images from human paraffin-embedded breast, cervical, ovarian and endometrial cancer sections, and IF images from human paraffin-embedded colon and endometrial cancer sections (catalog image captions).
Real IHC data
Anti-RPL36 Antibody ®
Cat # A08922-3
A08922-3 is the only rendered SKU; its IHC images show human paraffin-embedded breast, cervical, ovarian and endometrial cancer tissue (catalog IHC captions). Its IF images show human paraffin-embedded colon and endometrial cancer tissue (catalog IF captions).
Which to pick: For tissue IHC, choose A08922-3: its rabbit antibody was used on a human paraffin-embedded breast cancer section with EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (catalog IHC caption); the fixative is unreported (catalog IHC caption). For IF, the same SKU has paraffin-section images using 5 μg/ml primary antibody (catalog IF captions); ICC validation is unreported (catalog application list and IF captions). For cross-species planning, A08922-3 lists human, monkey, mouse and rat reactivity (catalog reactivity), while its IHC-P dilution entry specifies human and rat (catalog dilution line).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9Y3U8 (RL36_HUMAN, Large ribosomal subunit protein eL36).
- Human Protein Atlas. RPL36 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. RPL36 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. RPL36 antibody validation summary (1 antibodies).
- LncRNA PLAC2 down-regulates RPL36 expression and blocks cell cycle progression in glioma through a mechanism involving STAT1. Journal of cellular and molecular medicine 2018 — PMC5742712.
- PubMed PMID:11875025 — UniProt-cited evidence.
- PubMed PMID:11230166 — UniProt-cited evidence.
- PubMed PMID:11042152 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RPL36 (Large ribosomal subunit protein eL36). Expected localisation General cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A08922-3). Validated IHC antibody: A08922-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Endocrine cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A08922-3); 2. Fixation: Image fixative and duration unreported (datasheet A08922-3); 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 A08922-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08922-3); 8. Primary antibody: Rabbit anti-RPL36, 2-5 μg/ml (datasheet A08922-3); 9. Primary incubation: Overnight at 4 °C (datasheet A08922-3). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Adipocytes and ovarian stroma show low staining (HPA tissue IHC); Isoform / epitope: No isoforms annotated; mature chain spans residues 2–105 (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt Q9Y3U8, HPA tissue IHC · Supported, HPA ICC-IF · cytosol., Datasheet A08922-3, PMC5742712. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RPL36/immunohistochemistry-ihc-guide.