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RPL17 / Large ribosomal subunit protein uL22 · IHC design guide
Design Immunohistochemistry for RPL17
Plan RPL17 chromogenic IHC in paraffin sections using the catalog antibody at 2–5 μg/ml (datasheet A06980-2). Compare cytoplasmic staining with the HPA tissue IHC profile, including high staining in colon glandular cells (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RPL17 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 (HPA tissue IHC) | |
| Staining pattern | Ubiquitous cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06980-2) | |
| Positive control | Adrenal gland+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 paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Staining may detect products of more than one gene (HPA tissue IHC) | |
| Regulation | No regulatory driver reported (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope effects unknown (UniProt) |
Section 1
Recommended RPL17 IHC & IF Protocols
The catalog antibody protocol uses heat retrieval with EDTA pH 8.0 (datasheet A06980-2). Published IHC protocols cover zebra finch brain, mouse carotid arteries, and rat and clinical tissue (PMC2991423; PMC3502943; PMC9117278).
| Sample | Paraffin-embedded human thyroid papillary carcinoma tissue; fixative not specified (datasheet A06980-2) |
| Fixation | Image fixative and duration unreported (datasheet A06980-2); 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 A06980-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06980-2) |
| Primary antibody | Rabbit anti-RPL17, 2-5 μg/ml (datasheet A06980-2) |
| Primary incubation | Overnight at 4 °C (datasheet A06980-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A06980-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RPL17-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A06980-2). Use 10 mM citrate, pH 6.0 when reproducing the mouse carotid protocol (PMC3502943).
Section 2
What Is the Expected RPL17 Staining Pattern?
RPL17 is a cytoplasmic ribosomal protein with no transmembrane segment (UniProt P18621). In paraffin-section IHC, expect cytoplasmic staining in many cell types, including colon glandular cells, bronchial respiratory epithelial cells, and cerebral cortical neurons with reported high staining (HPA tissue IHC). Treat the pattern as provisional: HPA rates its tissue IHC Approved with medium RNA consistency, pending external verification, and cautions that the antibodies target proteins from more than one gene (HPA tissue IHC).
| Cytoplasmic staining in colon glandular cells, bronchial respiratory epithelial cells, or cortical neurons. | This fits the reported high-staining cell populations and ubiquitous cytoplasmic profile (HPA tissue IHC). Judge the slide by both compartment and cell identity; a strong chromogen signal alone does not establish RPL17 specificity, given HPA's multiple-gene targeting caution (HPA tissue IHC). |
| Predominantly nuclear staining, with little cytoplasmic signal in an expected positive population. | A nuclear-dominant result conflicts with UniProt's cytoplasmic annotation and the HPA tissue profile (UniProt P18621; HPA tissue IHC). Consider an off-target or detection artefact before scoring it as RPL17; compare the staining with the expected positive cells and a negative detection control (general IHC practice). |
| Strong staining in a cell population reported as low, such as smooth muscle cells or placental trophoblastic cells. | HPA reports low staining in those populations, so unexpectedly strong signal warrants a specificity check (HPA tissue IHC). It may reflect antibody cross-reactivity or endogenous detection activity; the image alone cannot distinguish these causes, especially with HPA's multiple-gene targeting caution (HPA tissue IHC; general IHC practice). |
| Diffuse chromogen across tissue, empty spaces, or multiple compartments without a clear cellular pattern. | This is difficult to reconcile with the reported cytoplasmic pattern (HPA tissue IHC). Assess background with a primary-antibody omission control, then review blocking, washing, and detection conditions; these are general IHC checks, not documented RPL17-specific sensitivities (general IHC practice). |
| No cytoplasmic signal in an adequately represented colon glandular or bronchial epithelial population. | Those populations are reported high by HPA, making an absent signal worth investigating (HPA tissue IHC). First confirm that tissue morphology and detection controls are interpretable; then review the catalog antibody's IHC-P conditions. HPA's Approved rating does not guarantee staining in every preparation (HPA tissue IHC; general IHC practice). |
💡Expected RPL17 appearanceCall positive IHC when expected glandular, respiratory epithelial, or neuronal cells show discernible cytoplasmic staining, with high levels reported in the named examples; isolated nuclear signal or noncellular chromogen is suspect (HPA tissue IHC; UniProt P18621; general IHC practice).
| Which compartments should guide scoring? | UniProt annotates RPL17 as cytoplasmic, and HPA describes ubiquitous cytoplasmic tissue staining (UniProt P18621; HPA tissue IHC). Score cellular cytoplasm rather than a nuclear-only pattern; HPA ICC-IF also supports cytosol and endoplasmic reticulum localization, but that assay does not define a separate IHC compartment (HPA subcellular). |
| Which tissue gives an interpretable positive comparison? | HPA reports high staining in colon glandular cells, bronchial respiratory epithelial cells, and cerebellar Purkinje cells, among others (HPA tissue IHC). A comparison should include an identifiable reported-positive cell population; HPA also lists low staining in smooth muscle cells and ovarian stroma cells (HPA tissue IHC). |
| How certain is antibody specificity? | Two rabbit polyclonal antibodies, HPA043724 and HPA046385, are IHC Approved, while the tissue assessment reports medium RNA consistency, pending external verification, and targeting of proteins from more than one gene (HPA antibodies; HPA tissue IHC). Interpret a plausible pattern as supportive, not definitive proof of RPL17 identity. |
| Do isoforms or processing predict a different slide pattern? | UniProt lists three isoforms and a mature chain spanning residues 2–184, with no signal peptide, propeptide, or transmembrane segment (UniProt P18621). These annotations do not establish isoform-specific staining or a different tissue compartment; assess the observed IHC pattern against the cytoplasmic annotation (UniProt P18621; HPA tissue IHC). |
| Does IF/ICC offer a localization cross-check? | HPA ICC-IF mainly localizes the signal to endoplasmic reticulum and cytosol and labels both locations supported (HPA subcellular). Its multiple-gene targeting caution still applies; use this as a compartment comparison, while keeping IF/ICC conditions with the separate IF/ICC guide (HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected-positive tissue has no visible cytoplasmic staining. | The result conflicts with HPA's high staining in the selected cell population, but does not identify a single failed step (HPA tissue IHC). | Confirm the relevant cells are present, check detection controls, and review the catalog antibody's IHC-P conditions, including its stated retrieval and dilution where available (general IHC practice). |
| Signal is nuclear-dominant in otherwise intact cells. | The compartment conflicts with the cytoplasmic annotations; off-target or detection artefact is possible (UniProt P18621; HPA tissue IHC). | Compare with a reported-positive cytoplasmic population and a primary-antibody omission control before assigning RPL17 positivity (HPA tissue IHC; general IHC practice). |
| Low-reported cells stain more strongly than nearby expected-positive cells. | The pattern conflicts with HPA's low and high cell-population entries; cross-reactivity or endogenous detection activity may contribute (HPA tissue IHC; general IHC practice). | Check cell identity and compare appropriate omission and detection controls. Treat the result as unresolved if those controls do not explain it (general IHC practice). |
| Chromogen appears diffusely over the section or in spaces between cells. | Noncellular background does not match HPA's cytoplasmic cellular profile (HPA tissue IHC). Excess detection background is one possible general IHC cause (general IHC practice). | Inspect the primary-antibody omission control, then review blocking, washing, and detection settings under the established IHC workflow (general IHC practice). |
| Staining varies between candidate positive tissues. | HPA reports high levels in specified cell populations but also low levels in others; its tissue IHC assessment has medium RNA consistency (HPA tissue IHC). | Compare the same identified cell types and compartments across sections, and document which populations were scored (HPA tissue IHC; general IHC practice). |
| A plausible cytoplasmic pattern is being treated as definitive RPL17 identification. | HPA notes multiple-gene targeting and pending external verification despite an Approved tissue IHC rating (HPA tissue IHC). | Report the pattern and antibody identifier with the specificity caveat; seek independent evidence before making a target-specific conclusion (HPA antibodies; HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing RPL17 staining
A06980-2 · IHCIHC analysis of RPL17 using anti-RPL17 antibody (A06980-2). RPL17 was detected in a paraffin-embedded section of human thyroid papillary car
A06980-2 · IHCIHC analysis of RPL17 using anti-RPL17 antibody (A06980-2). RPL17 was detected in a paraffin-embedded section of human laryngeal carcinoma t
A06980-2 · IFIF analysis of RPL17 using anti-RPL17 antibody (A06980-2). RPL17 was detected in an immunocytochemical section of U2OS cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000215472-RPL17/tissue/cerebral+cortex#imid_11897279
HPA · Oral mucosaHuman Protein Atlas IHC, Oral mucosa. Source: https://www.proteinatlas.org/ENSG00000215472-RPL17/tissue/oral+mucosa#imid_11898931
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000215472-RPL17/tissue/colon#imid_11898917
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000215472-RPL17/tissue/liver#imid_11898918
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043724; green = RPL17, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043724; green = RPL17, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RPL17 IHC & IF
🧪Run adrenal gland first: its glandular cells should stain (HPA: High in adrenal gland glandular cells). HPA detects RPL17 in all 45 scored tissues, so there is no supported negative tissue or reliable internal negative cell population; use no-primary and isotype slides to judge background (HPA: no negative rows).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: RPL17 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 RPL17 in A-431, U-251MG, U2OS, HEK293, Hep-G2, with annotated localisation: Endoplasmic reticulum (supported), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide, a host-matched rabbit IgG isotype control, and RPL17 knockout material or a validated peptide-block control for specificity (caption: rabbit anti-RPL17 antibody). For chromogenic detection, quench endogenous peroxidase and check endogenous biotin background because the selected caption uses biotinylated secondary antibody and SABC detection (caption: biotinylated goat anti-rabbit IgG, SABC and DAB).
⚠️Feasibility: The selected tissue-IHC caption reports heat retrieval in EDTA at pH 8.0 before staining a paraffin section, but reports no fixative; a target-specific fixation window or fixation effect is unreported (selected-SKU tissue-IHC caption). Retrieval was used in that example, but whether it is required for every specimen and whether frozen sections or IF are easier is unreported (selected-SKU tissue-IHC caption; HPA: ICC-IF images available). On the adrenal positive slide, check for background from the biotin-based detection system before scoring glandular-cell staining (HPA: High in adrenal gland glandular cells; caption: SABC detection).
HPA tissue IHC evidence for RPL17
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification. Caution, targets protein from more than one gene.). 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 | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: RPL17 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced RPL17 IHC Tips
Use the catalog antibody’s paraffin-section IHC conditions as the starting point, then evaluate cytoplasmic staining with appropriate tissue and detection controls.
How should I optimize retrieval when RPL17 staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A06980-2). The selected tissue image used that retrieval before 2 μg/ml primary antibody overnight at 4°C, so reproduce those conditions before changing them (datasheet A06980-2). If staining remains weak, adjust heating duration in a controlled series while keeping section thickness, detection chemistry and primary concentration constant (standard IHC practice). Include a tissue with documented high cellular staining, such as colon glandular cells, alongside each condition (HPA: High in colon glandular cells). Compare cytoplasmic signal and tissue preservation; excessive retrieval can damage morphology and make apparent gains difficult to interpret (standard IHC practice).
Could fixation explain weak or uneven RPL17 staining?
The selected paraffin-section caption does not state a fixative, so target-specific fixation sensitivity is unknown (datasheet A06980-2: fixative not stated). Record the fixative, time in fixative and processing history for each specimen before comparing RPL17 staining across cases (standard IHC practice). If fixation varies, process a small matched series with the same EDTA pH 8.0 retrieval and detection conditions (datasheet A06980-2; standard IHC practice). Assess preserved morphology and cytoplasmic staining together, using an appropriate positive tissue and a primary-omission control (UniProt P18621: cytoplasm; standard IHC practice). Do not attribute a difference to RPL17 biology until specimen handling and staining variation have been assessed (standard IHC practice).
What staining pattern should count as plausible RPL17 signal?
Prioritize cytoplasmic staining in intact cells: RPL17 is a large ribosomal subunit component annotated in the cytoplasm (UniProt P18621: function and subcellular location). Tissue IHC reports ubiquitous cytoplasmic expression, with high staining in colon glandular cells and bronchial respiratory epithelial cells (HPA: tissue IHC profile and positive tissues). A predominantly membrane-rim pattern has no support from the annotated topology, which has no transmembrane segment (UniProt P18621: topology). Compare staining within comparable cell populations and against a primary-omission section before assigning faint diffuse colour to RPL17 (standard IHC practice). Review the hematoxylin counterstain to distinguish cellular cytoplasm from extracellular deposits or damaged tissue (standard IHC practice).
Can this IHC antibody distinguish RPL17 isoforms or a processed epitope?
RPL17 has 3 annotated isoforms, but the supplied tissue-IHC caption does not identify the antibody epitope or establish isoform selectivity (UniProt P18621: isoforms; datasheet A06980-2). Its annotated mature chain spans residues 2–184, with no signal peptide or propeptide reported (UniProt P18621: processing). No glycosylation sites or modified residues are annotated, so those records provide no basis for predicting a retrieval-sensitive modification at this antibody’s epitope (UniProt P18621: PTMs). Treat a positive chromogenic section as RPL17-reactive staining rather than an isoform assignment (standard IHC interpretation). For isoform-specific claims, first obtain the immunogen sequence and establish specificity with an appropriate independent control (standard antibody-validation practice).
How can I check RPL17 localisation with multiplex IF?
On the separate IF/ICC workflow, pair RPL17 with a marker for the expected cell population, such as an epithelial marker when examining glandular cells (HPA: High in colon glandular cells; standard IF practice). Choose fluorophores after checking the specimen’s autofluorescence, reserving a spectrally distinct channel for RPL17 and including single-stain controls (standard IF practice). Apply controlled permeabilisation so antibody can reach the cytoplasmic side of cellular membranes, then compare signal with a no-primary control (UniProt P18621: cytoplasm and no transmembrane segment; standard IF practice). HPA reports endoplasmic-reticulum and cytosol IF localisation but cautions that the underlying antibodies target proteins from multiple genes (HPA: subcellular summary). Do not treat that IF pattern as validation of this catalog antibody (HPA: subcellular caution).
How do I separate diffuse RPL17 staining from chromogenic background?
Because RPL17 is reported as ubiquitously cytoplasmic, diffuse cellular colour alone is a weak specificity test (HPA: tissue IHC profile). Run a primary-omission section to assess secondary and detection background, and include a peroxidase block when using DAB detection (standard IHC practice). The selected image used 10% goat serum blocking, a biotinylated secondary and streptavidin-biotin detection with DAB (datasheet A06980-2). For this detection system, check endogenous biotin and endogenous peroxidase as possible sources of colour (standard IHC practice). Reduce nonspecific staining through controlled washes and antibody titration while retaining interpretable cytoplasmic signal in a positive tissue (standard IHC practice; UniProt P18621: cytoplasm).
How should I score RPL17 across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic compartment before scoring, since RPL17 is annotated in the cytoplasm and tissue staining is broadly distributed (UniProt P18621: subcellular location; HPA: tissue IHC profile). For comparable specimens, record the percentage of positive intact cells and staining intensity, then calculate a cytoplasmic H-score on a 0–300 scale (standard IHC scoring practice). Normalize counts to the number of eligible cells in the same annotated region; for spatial counts, report positive cells per mm² of viable tissue (standard image-analysis practice). Keep exposure, section processing, retrieval and threshold settings consistent across the comparison (standard IHC practice). Exclude necrosis and tissue edges from the scored region and report the exclusion rule (standard IHC scoring practice).
When is an apparent RPL17-positive section likely misleading?
A convincing result shows cellular cytoplasmic staining in intact tissue and exceeds the matched detection-control signal (UniProt P18621: cytoplasm; standard IHC practice). Strong membrane-only colour is discordant with RPL17’s lack of a transmembrane segment, while colour concentrated at cut edges or necrosis warrants artefact review (UniProt P18621: topology; standard IHC practice). Check whether the coloured cells match the annotated region; HPA reports high staining in colon glandular cells but low staining in smooth muscle cells (HPA: positive and low tissues). With DAB detection, investigate colour that persists after primary omission as possible endogenous enzyme or detection-system signal (standard IHC practice). Interpret biological differences cautiously because HPA notes medium RNA–staining consistency and cautions that its antibodies target proteins from multiple genes (HPA: reliability description).
Boster reagents
Best RPL17 / Large ribosomal subunit protein uL22 IHC Antibodies
A06980-2 has IHC data from human paraffin sections and IF/ICC data from U2OS cells (catalog image captions); listed reactivity includes human, mouse and rat (catalog applications and reactivity).
Real IHC data
Anti-RPL17 Antibody ®
Cat # A06980-2
A06980-2 will render with IHC data from human thyroid papillary carcinoma in a paraffin section (A06980-2 IHC figure caption). Its additional IHC captions show human laryngeal, liver and stomach cancer sections, and its IF caption shows U2OS cells (A06980-2 image captions).
Which to pick: Choose A06980-2 for paraffin-section IHC: its figure used EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (A06980-2 IHC figure caption); the fixative is unreported (A06980-2 IHC figure caption). For IF/ICC, the same SKU has a U2OS-cell IF image using 5 μg/ml primary antibody (A06980-2 IF figure caption). For mouse or rat work, A06980-2 lists both species as reactive, although the supplied IHC and IF images show human samples only (catalog reactivity; A06980-2 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P18621 (RL17_HUMAN, Large ribosomal subunit protein uL22).
- Human Protein Atlas. RPL17 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. RPL17 subcellular location (ICC-IF): Mainly localized to the endoplasmic reticulum and cytosol. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. RPL17 antibody validation summary (2 antibodies).
- Sex- and age-related differences in ribosomal proteins L17 and L37, as well as androgen receptor protein, in the song control system of zebra finches. Neuroscience 2010 — PMC2991423.
- Effects of estradiol on incorporation of new cells in the developing zebra finch song system: potential relationship to expression of ribosomal proteins L17 and L37. Developmental neurobiology 2009 — PMC2692582.
- Ribosomal protein L17, RpL17, is an inhibitor of vascular smooth muscle growth and carotid intima formation. Circulation 2012 — PMC3502943.
- Comparative proteomic analysis identifies differentially expressed proteins and reveals potential mechanisms of traumatic heterotopic ossification progression. Journal of orthopaedic translation 2022 — PMC9117278.
- PubMed PMID:2402465 — UniProt-cited evidence.
- PubMed PMID:1793733 — UniProt-cited evidence.
- PubMed PMID:11875025 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RPL17 (Large ribosomal subunit protein uL22). Expected localisation Cytoplasmic staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06980-2). Validated IHC antibody: A06980-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular 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 thyroid papillary carcinoma tissue; fixative not specified (datasheet A06980-2); 2. Fixation: Image fixative and duration unreported (datasheet A06980-2); 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 A06980-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06980-2); 8. Primary antibody: Rabbit anti-RPL17, 2-5 μg/ml (datasheet A06980-2); 9. Primary incubation: Overnight at 4 °C (datasheet A06980-2). Watch for: Fixation: Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Staining may detect products of more than one gene (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope effects unknown (UniProt); Regulation: No regulatory driver reported (UniProt). Evidence: UniProt P18621, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the endoplasmic reticulum and cytosol. Caution:…, Datasheet A06980-2, PMC2991423, PMC3502943. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RPL17/immunohistochemistry-ihc-guide.