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RPL27A / Large ribosomal subunit protein uL15 · IHC design guide
Design Immunohistochemistry for RPL27A
RPL27A is a cytoplasmic component of the large ribosomal subunit (UniProt). This guide covers paraffin-section IHC planning and interpretation using the cytoplasmic staining profile reported across most tissues (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RPL27A 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 most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic signal in most tissues, including glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Skeletal muscle+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07864-1) | |
| Caveat | Medium staining–RNA concordance; verification pending (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | No isoforms annotated; one chain spans residues 1–148 (UniProt) |
Section 1
Recommended RPL27A IHC & IF Protocols
The catalog antibody's IHC-P protocol is paired with a published RPL27A tissue microarray protocol using HCC and peritumoral liver samples (PMC10360225).
| Sample | Paraffin-embedded human colorectal carcinoma tissue; fixative not specified (datasheet A07864-1) |
| Fixation | Image fixative and duration unreported (datasheet A07864-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-RPL27A, 1:50-1:200 (datasheet A07864-1) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RPL27A-positive staining in purkinje cells - cytoplasm/membrane of cerebellum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min for this cytoplasmic target (page antigen retrieval; UniProt P46776: cytoplasm).
Section 2
What Is the Expected RPL27A Staining Pattern?
RPL27A is a cytoplasmic component of the large ribosomal subunit with no transmembrane segment (UniProt P46776). In paraffin tissue IHC, expect cytoplasmic staining across many tissues, especially the listed high-staining glandular cells and cerebellar Purkinje cells (HPA tissue IHC). HPA rates the tissue pattern Supported, with medium agreement between staining and RNA data and external verification pending (HPA tissue IHC).
| Glandular cytoplasm stains strongly in endometrium, pancreas, or salivary gland. | This matches HPA High staining in endometrial glandular cells, pancreatic exocrine glandular cells, and salivary glandular cells (HPA tissue IHC). Assess the named cells and their cytoplasm when judging whether a positive control has worked. |
| Purkinje cells show strong cytoplasmic staining, sometimes with a membrane pattern. | HPA records High staining in Purkinje cells as cytoplasm/membrane (HPA tissue IHC). A membrane component in these cells can fit that observation; interpret it alongside the expected cytoplasmic signal (UniProt P46776; HPA tissue IHC). |
| Signal is predominantly nuclear, or confined to cell borders outside the described Purkinje pattern. | That distribution does not match UniProt's cytoplasmic localisation or HPA's broad tissue profile (UniProt P46776; HPA tissue IHC). Treat it as a possible staining artefact and check the control slide before assigning RPL27A positivity. |
| Myocytes stain strongly while nearby expected positive cells do not. | HPA reports RPL27A as Not detected in skeletal myocytes and smooth muscle cells (HPA tissue IHC). Such a reversal raises concern for antibody cross-reactivity or endogenous chromogenic activity; the pattern alone cannot distinguish their causes. |
| Diffuse colour obscures cell boundaries, or a listed High tissue has no detectable signal. | Diffuse colour limits compartment scoring; absent signal in a listed positive tissue calls for a technical check (HPA tissue IHC). Because tissue IHC support has only medium staining/RNA consistency, one discordant section is not definitive evidence of target absence (HPA tissue IHC). |
💡Expected RPL27A appearanceCall a section positive when the expected glandular or Purkinje cells show clear cytoplasmic staining, often High in the listed reference tissues; diffuse background or dominant nuclear colour is suspect (HPA tissue IHC; UniProt P46776).
| Cellular compartment | UniProt places RPL27A in the cytoplasm and identifies it as a large ribosomal subunit component (UniProt P46776). Score cell-associated cytoplasmic colour first; HPA also records cytoplasm/membrane staining specifically in Purkinje cells (HPA tissue IHC). |
| Reference tissue and cell type | HPA lists High staining in endometrial, pancreatic exocrine, and salivary glandular cells, but Not detected in skeletal and smooth muscle cells (HPA tissue IHC). Compare like cell types within each section; a whole-tissue average can hide the relevant pattern. |
| Strength of tissue evidence | Two listed rabbit polyclonal antibodies, HPA060776 and HPA063379, have Supported IHC status (HPA antibodies). The tissue profile itself has medium agreement with RNA data and awaits external verification (HPA tissue IHC), so use discordance as a reason to investigate. |
| IF/ICC: where should signal appear? | In the separate IF/ICC application, HPA reports enhanced localisation to endoplasmic reticulum and cytosol; both listed antibodies have Enhanced ICC status (HPA subcellular; HPA antibodies). This observation helps interpret fluorescence images without changing the tissue IHC scoring reference. |
| Chromogenic detection controls | Endogenous enzyme activity and nonspecific reagent binding can contribute colour in chromogenic IHC (standard IHC practice). Include an appropriate negative detection control and examine whether colour follows the expected cell and compartment pattern; these controls do not establish target specificity by themselves. |
| Situation | Likely cause | Next action |
|---|---|---|
| No cytoplasmic colour in a listed High control tissue. | The run may have failed, or the sampled section may differ from HPA's observed pattern (HPA tissue IHC). | Check section integrity, retrieval and detection steps, then repeat with an appropriate positive control (standard IHC practice). Interpret persistent absence cautiously because HPA tissue IHC has medium staining/RNA consistency. |
| Colour appears across the slide, including spaces without cells. | Diffuse reagent background or chromogen deposition can obscure the expected cytoplasmic pattern (standard IHC practice). | Review blocking, washes, detection reagents and a negative detection control; score RPL27A only where cell-associated colour can be assessed against HPA's cytoplasmic profile (HPA tissue IHC). |
| Skeletal or smooth muscle cells stain strongly. | HPA records these cells as Not detected; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; standard IHC practice). | Inspect the negative detection control and compare an expected positive cell population in the same run. Do not assign specificity from muscle staining alone. |
| Most colour is nuclear. | A dominant nuclear pattern conflicts with the reported cytoplasmic location (UniProt P46776; HPA tissue IHC). | Recheck counterstain and chromogen interpretation, inspect control slides, and reassess whether distinct cytoplasmic colour is present before calling the section positive. |
| Purkinje cell borders show colour alongside cytoplasm. | HPA describes High Purkinje staining as cytoplasm/membrane (HPA tissue IHC). | Check that cytoplasmic staining accompanies the border signal, then compare with other expected cells. Border colour alone cannot establish specificity. |
| IHC appears broadly cytoplasmic, while an IF/ICC image looks more structured. | HPA tissue IHC reports cytoplasmic expression in most tissues, whereas HPA ICC-IF places RPL27A in endoplasmic reticulum and cytosol (HPA tissue IHC; HPA subcellular). | Interpret each image using its application-specific HPA localisation. Use cytoplasmic cell staining for the paraffin IHC call; consult the separate IF/ICC guide for fluorescence experiments. |
Experiment evidence
IHC & IF images showing RPL27A staining
A07864-1 · IHCImmunohistochemistry (IHC) analyzes of RPL27A (V140) pAb in paraffin-embedded human colorectal carcinoma tissue at 1:50.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000166441-RPL27A/tissue/cerebral+cortex#imid_17771139
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000166441-RPL27A/tissue/colon#imid_17771057
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000166441-RPL27A/tissue/liver#imid_17771050
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000166441-RPL27A/tissue/pancreas#imid_17771009
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA060776; green = RPL27A, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA060776; green = RPL27A, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RPL27A IHC & IF
🧪Run cerebellum first and look for staining in Purkinje cells (HPA: High in Purkinje cells, cytoplasm/membrane). Run skeletal muscle as the negative tissue (HPA: myocytes Not detected); use unstained cells within the cerebellum section as a background reference without assuming they are target-negative.
Positive control tissue: Cerebellum (Purkinje cells - cytoplasm/membrane, HPA High)
Negative control tissue: Skeletal muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RPL27A in HEK293, Hep-G2, U2OS, A-549, with annotated localisation: Endoplasmic reticulum (enhanced), Cytosol (enhanced) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host- and clonality-matched isotype controls, plus RPL27A knockout material or cognate peptide competition if available. Block endogenous peroxidase for chromogenic IHC; for cerebellar IF, check unstained tissue for neuronal lipofuscin autofluorescence (HPA: cerebellar Purkinje cells High).
⚠️Feasibility: No target-specific fixation window, fixation effect, or antigen-retrieval requirement is reported in the supplied evidence. The selected SKU caption shows paraffin-section IHC in human colorectal carcinoma at 1:50, but its fixative is unreported (selected SKU A07864-1 caption). ICC-IF images exist for HEK293, Hep-G2, U2OS and A-549 (HPA: subcellular); the evidence does not establish that frozen sections or IF are easier than paraffin IHC.
HPA tissue IHC evidence for RPL27A
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebellum | Purkinje cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Pancreas | Exocrine glandular cells | High | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | High | Protein (IHC) | HPA → |
| Stomach | Glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced RPL27A IHC Tips
Troubleshoot RPL27A staining in paraffin sections by checking retrieval, cytoplasmic distribution, tissue context and control staining before scoring.
How should I retrieve RPL27A in paraffin sections when staining is weak?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval rule: cytoplasmic/membrane antigen). Keep cooling, wash conditions and chromogenic development consistent across test and control sections so a retrieval change can be judged fairly (standard IHC practice). If staining remains weak, test a second retrieval condition on adjacent sections while retaining the citrate condition as the reference, and assess tissue preservation alongside signal (standard IHC practice). The catalog image documents RPL27A staining in paraffin-embedded human colorectal carcinoma at 1:50, but its caption does not report a fixative or retrieval method (selected IHC caption: A07864-1).
Could fixation explain weak or uneven RPL27A staining?
Target-specific fixation sensitivity is unknown because the supplied RPL27A tissue-IHC caption does not state a fixative (selected IHC caption: A07864-1). Record fixation history and compare sections processed together before attributing a weak signal to the antigen or antibody (standard IHC practice). Apply the page’s citrate pH 6.0 retrieval at 95–98 °C for 20 min consistently, then examine both staining and tissue integrity (page retrieval rule: cytoplasmic/membrane antigen; standard IHC practice). Do not infer a fixation effect from cytoplasmic expression, ribosomal function or annotated modifications; those observations do not test fixation sensitivity (HPA: cytoplasmic expression in most tissues; UniProt P46776: large ribosomal subunit component and modified residues).
Where should credible RPL27A chromogenic staining appear?
Expect predominantly cytoplasmic staining in tissue sections (HPA: cytoplasmic expression in most tissues; UniProt P46776: cytoplasm). An endoplasmic reticulum associated or diffuse cytosolic pattern is biologically plausible, although that finer assignment comes from ICC/IF evidence rather than the tissue-IHC profile (HPA subcellular: enhanced endoplasmic reticulum and cytosol). RPL27A has no annotated transmembrane segment, so a crisp plasma membrane rim alone needs scrutiny (UniProt P46776: topology). Compare the pattern with adjacent tissue and a no-primary control, using the same hematoxylin counterstain and chromogenic development conditions (standard IHC practice). Score cytoplasmic signal separately from any isolated nuclear, luminal or edge staining rather than combining them into one intensity value (standard IHC practice).
Can isoforms or modified residues alter how I interpret staining?
The supplied record lists 0 isoforms and a single 1–148 chain for RPL27A (UniProt P46776: isoforms and processing). It also lists modified residues, including hydroxyhistidine 39, acetyllysines 47, 55 and 110, and phosphoserine 68 (UniProt P46776: modified residues). Without a supplied antibody epitope, none of those sites establishes a cause of weak staining or proves that a particular modification changes binding (selected IHC caption: no epitope reported). If results differ across preparations, compare matched sections under the same citrate pH 6.0 retrieval and detection conditions, and treat any epitope explanation as a hypothesis pending direct validation (page retrieval rule; standard IHC practice).
How can IF help check an ambiguous RPL27A IHC pattern?
Use IF as a complementary localisation check: HPA reports RPL27A in the endoplasmic reticulum and cytosol in ICC/IF images (HPA subcellular: enhanced endoplasmic reticulum and cytosol). Multiplex with a validated marker for the cell type being assessed, selecting spectrally separated fluorophores and a channel with low tissue autofluorescence after checking unstained sections (standard IF practice). Because RPL27A has no annotated transmembrane segment, permeabilisation should allow antibody access to the intracellular epitope; optimise its strength against cell morphology and the marker signal (UniProt P46776: topology; standard IF practice). Keep the paraffin-section result anchored to its own controls and documented 1:50 catalog image, since that caption supplies no IF protocol (selected IHC caption: A07864-1).
What should I check when diffuse brown staining obscures RPL27A?
First inspect a no-primary section for chromogen deposition and compare its distribution with the test section (standard IHC practice). For peroxidase detection, include an endogenous peroxidase block and check whether the chosen detection system needs a biotin-related control; these are general chromogenic safeguards, not RPL27A-specific findings (standard IHC practice). Titrate the primary antibody against background using the catalog image’s 1:50 dilution as documented context, without assuming it is optimal for every preparation (selected IHC caption: A07864-1; standard IHC practice). Examine folds, cut edges and necrotic areas separately, and require a reproducible cytoplasmic cellular pattern before calling a diffuse brown field positive (HPA: cytoplasmic tissue expression; standard IHC practice).
How should I score RPL27A across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored compartment as cytoplasm before reviewing cases, consistent with the reported tissue pattern (HPA: cytoplasmic expression in most tissues). For a cell population with clear boundaries, report the percentage of positive cells and an intensity-based H-score; record the scoring thresholds and keep them fixed across sections (standard IHC practice). Where cell segmentation is unreliable, report positive-cell density per mm² of viable, annotated tissue and state the counting rule (standard IHC practice). Normalise comparisons to the same cell type, viable tissue area, staining batch and exposure to chromogenic development, rather than comparing raw brown area across unlike regions (standard IHC practice).
When is a strong RPL27A signal convincing rather than artefactual?
A convincing result shows reproducible cellular cytoplasmic staining in intact tissue, with low signal in the matching no-primary control (HPA: cytoplasmic expression in most tissues; standard IHC practice). High staining in pancreatic exocrine glandular cells or cerebellar Purkinje cells fits reported tissue examples, while skeletal and smooth muscle cells were reported as not detected (HPA: listed tissue-IHC cell populations). Treat isolated nuclear signal, a sharp cut-edge gradient, staining confined to necrosis or brown deposits reproduced without primary antibody as reasons to investigate the section or detection chemistry (standard IHC practice). HPA rates its tissue-IHC evidence as Supported but notes medium RNA agreement and pending external verification, so use matched controls when making a biological claim (HPA: reliability description).
Boster reagents
Best RPL27A / Large ribosomal subunit protein uL15 IHC Antibodies
One anti-RPL27A antibody has real IHC data from paraffin-embedded human colorectal carcinoma; its listed reactivity includes human, mouse, and rat, with no IF data supplied (catalog: A07864-1).
Real IHC data
Anti-RPL27A (V140) Antibody
Cat # A07864-1
A07864-1 has an IHC image of paraffin-embedded human colorectal carcinoma stained at 1:50 (image caption: A07864-1). It is listed for IHC and reacts with human, mouse, and rat; no IF image is supplied (catalog: A07864-1).
Which to pick: For paraffin-section tissue IHC, choose A07864-1: its own image shows human colorectal carcinoma at 1:50, within the listed 1:50–1:200 IHC range; the fixative is unreported (image caption: A07864-1; catalog: A07864-1). No SKU in this payload is listed for IF/ICC (catalog: A07864-1 applications). For mouse or rat tissue IHC, A07864-1 lists reactivity with both species, though its IHC image shows human tissue only; it is rabbit-hosted, and its caption identifies it as a pAb (catalog: A07864-1; image caption: A07864-1).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P46776 (RL27A_HUMAN, Large ribosomal subunit protein uL15).
- Human Protein Atlas. RPL27A tissue IHC expression (reliability: Supported).
- Human Protein Atlas. RPL27A subcellular location (ICC-IF): Localized to the endoplasmic reticulum and cytosol..
- Human Protein Atlas. RPL27A antibody validation summary (2 antibodies).
- High expression of RPL27A predicts poor prognosis in patients with hepatocellular carcinoma. World journal of surgical oncology 2023 — PMC10360225.
- PubMed PMID:7772601 — UniProt-cited evidence.
- PubMed PMID:10449908 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RPL27A (Large ribosomal subunit protein uL15). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A07864-1 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells - cytoplasm/membrane); negative Skeletal muscle. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colorectal carcinoma tissue; fixative not specified (datasheet A07864-1); 2. Fixation: Image fixative and duration unreported (datasheet A07864-1); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-RPL27A, 1:50-1:200 (datasheet A07864-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07864-1); Caveat: Medium staining–RNA concordance; verification pending (HPA tissue IHC); Isoform / epitope: No isoforms annotated; one chain spans residues 1–148 (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). Evidence: UniProt P46776, HPA tissue IHC · Supported, HPA ICC-IF · endoplasmic reticulum and cytosol., Datasheet A07864-1, PMC10360225. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RPL27A/immunohistochemistry-ihc-guide.