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RPLP2 / Large ribosomal subunit protein P2 · IHC design guide
Design Immunohistochemistry for RPLP2
Plan RPLP2 paraffin IHC around general cytoplasmic staining and strong signal in kidney tubule cells (HPA tissue IHC). Start the catalog antibody at 1:100–1:300 (datasheet), and compare with adipocytes or myocytes, where staining was not detected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RPLP2 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 staining (HPA tissue IHC) | |
| Staining pattern | Kidney tubule cells: high, generally cytoplasmic signal (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Kidney+4 more · see all | |
| Negative control | Adipose tissue+3 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent (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 A05244-1) | |
| Caveat | Adipocytes and myocytes have no detected signal (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | No isoforms; one 1–115 chain (UniProt) |
Section 1
Recommended RPLP2 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with three published RPLP2 IHC methods (PMC12189634; PMC10697958; PMC5911611).
| Sample | Paraffin-embedded human prostate carcinoma tissue; fixative not specified (datasheet A05244-1) |
| Fixation | Image fixative and duration unreported (datasheet A05244-1); 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-RPLP2, 1:100 - 1:300 (datasheet A05244-1) |
| 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 | RPLP2-positive staining in cells in tubules of kidney (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 at 95–98 °C for 20 min (page antigen-retrieval setting); one published method instead used high-pH retrieval (PMC5911611).
Section 2
What Is the Expected RPLP2 Staining Pattern?
In paraffin-section IHC, expect predominantly cytoplasmic RPLP2 staining, with kidney tubular cells a useful strong reference and several epithelial and neuronal populations showing moderate staining (HPA: general cytoplasmic expression; kidney tubules High; listed populations Medium). The protein has no transmembrane segment and is part of the ribosomal stalk (UniProt P05387 topology and subunit). HPA rates its tissue IHC profile Approved, pending external verification (HPA: reliability).
| Kidney tubular cells show strong cytoplasmic chromogen, while nearby adipocytes lack convincing signal (HPA: kidney tubules High; adipocytes Not detected). | This supports the expected cell and compartment pattern on the slide. Judge staining in identifiable cells rather than treating all tissue as equally positive (HPA: general cytoplasmic expression; kidney tubules High; adipocytes Not detected). |
| Signal is predominantly nuclear or confined to cell borders, with little cytoplasmic staining in otherwise positive kidney tubules (HPA: kidney tubules High; general cytoplasmic expression). | Review this IHC result for nonspecific staining or a processing artefact before scoring it as RPLP2. Nuclear speckles are reported by ICC-IF, so their presence in that assay does not make broad nuclear chromogen the expected tissue IHC pattern (HPA: subcellular ICC-IF; tissue IHC profile). |
| Adipocytes, skeletal myocytes, smooth muscle cells, or chondrocytes stain as strongly as kidney tubules (HPA: these cells Not detected; kidney tubules High). | That distribution conflicts with the reported tissue IHC profile. Consider antibody cross-reactivity or endogenous detection activity; confirm cell identity and assess controls before calling those cells positive (HPA: tissue IHC levels; standard IHC practice). |
| Chromogen coats stroma, vessel lumens, and cell-free areas without a discernible cellular cytoplasmic pattern (HPA: general cytoplasmic expression). | Treat this as background rather than RPLP2 expression. Review blocking, washes, chromogen development, and section quality using the same run's controls (standard IHC practice). |
| Kidney tubular cells show no convincing staining in a run expected to detect RPLP2 (HPA: kidney tubules High). | The absent reference signal makes a negative result elsewhere difficult to interpret. Check the positive-control section and detection run, then review antibody conditions and retrieval before drawing a biological conclusion (HPA: kidney tubules High; standard IHC practice). |
💡Expected RPLP2 appearanceCall an IHC result consistent with RPLP2 when kidney tubular cells show strong, predominantly cytoplasmic staining, while conspicuous cell-free chromogen or equally strong adipocyte staining prompts a false-positive check (HPA: general cytoplasmic expression; kidney tubules High; adipocytes Not detected; standard IHC practice).
| Tissue and cell choice (HPA: tissue IHC) | Kidney tubules are reported High; glandular cells in adrenal gland, appendix, and breast are Medium, while adipocytes are Not detected. Score the identified cell population, since an overall tissue score can obscure this difference (HPA: listed tissue IHC levels). |
| Evidence strength (HPA: reliability and antibodies) | The tissue profile is Approved but pending external verification. HPA053635 has Approved IHC status; HPA075365 has Approved ICC status, which does not itself validate that antibody for paraffin-section IHC (HPA: tissue reliability; antibody statuses). |
| IF/ICC localisation? (HPA: subcellular ICC-IF) | In the separate IF/ICC guide, expect cytosol and nuclear speckles as approved main locations, with mitotic spindle as an additional location. These cell-image observations inform interpretation but are not an IHC-P protocol option (HPA: approved ICC-IF locations). |
| Protein context (UniProt P05387) | RPLP2 is a 115-residue ribosomal-stalk protein with no transmembrane segment, signal peptide, propeptide, or annotated isoforms. Those annotations support an intracellular interpretation but do not identify this antibody's epitope or predict antigen-retrieval behavior (UniProt P05387 topology, processing, isoforms, subunit). |
| Target-specific fixation sensitivity (HPA; UniProt P05387) | No target-specific fixation effect is supplied by the HPA tissue profile or UniProt record. Interpret retrieval changes as assay optimisation findings, without claiming that fixation alters RPLP2 epitope access or expression (HPA: tissue IHC scope; UniProt P05387 annotations). |
| Situation | Likely cause | Next action |
|---|---|---|
| Kidney tubular positive control is blank (HPA: kidney tubules High). | The run may have failed through antibody, retrieval, or detection conditions; the blank slide alone cannot identify which step failed (standard IHC practice). | Check control-section quality and reagent sequence, then compare a documented retrieval condition and antibody dilution on matched sections before interpreting study negatives (standard IHC practice). |
| All tissue, including cell-free areas, develops chromogen (standard IHC practice). | Excess detection signal, insufficient washing or blocking, or endogenous enzyme activity may contribute (standard IHC practice). | Review the no-primary control, detection chemistry, blocking, washes, and development time; require a cellular pattern before assigning RPLP2 positivity (standard IHC practice; HPA: general cytoplasmic expression). |
| Adipocytes or muscle cells appear strongly positive (HPA: adipocytes and skeletal and smooth muscle cells Not detected). | Cell misidentification, nonspecific antibody binding, or endogenous detection activity are possibilities; HPA's profile alone cannot distinguish them (HPA: tissue IHC levels; standard IHC practice). | Recheck morphology and a no-primary control, then compare with kidney tubules in the same run before accepting the unexpected signal (HPA: kidney tubules High; standard IHC practice). |
| Most signal is nuclear in paraffin-section IHC (HPA: general cytoplasmic tissue IHC expression). | Nonspecific nuclear chromogen or staining artefact may be present; approved ICC-IF nuclear speckles do not establish a predominantly nuclear tissue IHC pattern (HPA: tissue IHC; subcellular ICC-IF). | Inspect cellular detail and controls, then score cytoplasmic staining separately from nuclear signal rather than merging them into one positive call (HPA: tissue IHC profile; standard IHC practice). |
| Moderate reference tissues seem weaker than kidney tubules (HPA: listed Medium populations; kidney tubules High). | This can match the reported cell-level pattern, provided staining remains interpretable and controls pass (HPA: tissue IHC levels; standard IHC practice). | Score each identified population against its reported level and the run's kidney reference; avoid requiring equal intensity across tissues (HPA: tissue IHC levels; standard IHC practice). |
| Signal varies after changing retrieval conditions (standard IHC practice). | The assay conditions may change the observed staining; target-specific RPLP2 fixation sensitivity is unreported in the supplied HPA and UniProt evidence (HPA: tissue IHC scope; UniProt P05387 annotations). | Compare matched sections with positive and no-primary controls, document the condition that yields specific cytoplasmic staining, and avoid attributing the difference to a proven RPLP2 fixation effect (standard IHC practice; HPA: general cytoplasmic expression). |
Experiment evidence
IHC & IF images showing RPLP2 staining
A05244-1 · IHCImmunohistochemistry analysis of paraffin-embedded human prostate carcinoma tissue, using RPLP2 Antibody. The picture on the right is blocke
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000177600-RPLP2/tissue/cerebral+cortex#imid_16692111
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000177600-RPLP2/tissue/colon#imid_16691837
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000177600-RPLP2/tissue/liver#imid_16691822
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000177600-RPLP2/tissue/kidney#imid_16691839
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA075365; green = RPLP2, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA075365; green = RPLP2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RPLP2 IHC & IF
🧪Run kidney first: cells in tubules should stain (HPA: High in kidney cells in tubules). Use adipose tissue as the negative tissue: adipocytes are listed as not detected (HPA: Not detected in adipocytes); the supplied HPA rows identify no specific internal negative cell type within kidney, so do not designate one by appearance alone.
Positive control tissue: Kidney (Cells in tubules, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RPLP2 in HEK293, PC-3, U2OS, with annotated localisation: Nuclear speckles (approved), Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) slide and a host- and clonality-matched isotype control (standard IHC practice). Use a validated RPLP2 knockout sample or a peptide-block control as a biological specificity check (selected A05244-1 tissue-IHC caption: staining blocked with synthesized peptide); quench endogenous peroxidase and assess background in kidney tubules before interpreting chromogenic staining (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and the effect of fixation on RPLP2 staining are unreported in the supplied evidence; the selected A05244-1 paraffin-section caption does not state a fixative (selected A05244-1 tissue-IHC caption: fixative unreported). Retrieval dependency is unreported, so assess retrieval conditions empirically (supplied target/application evidence: no retrieval comparison). The supplied evidence does not establish whether frozen sections or IF are easier than paraffin IHC; kidney tubular endogenous peroxidase can complicate chromogenic interpretation (standard IHC practice).
HPA tissue IHC evidence for RPLP2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular 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 → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Chondrocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RPLP2 IHC Tips
Troubleshoot RPLP2 staining in paraffin sections using the catalog antibody’s tissue image, HPA expression patterns, and standard IHC practice.
How should I retrieve RPLP2 antigen when paraffin sections stain weakly?
Start with citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval setting). Run a known positive section alongside the test section; kidney tubule cells offer a high-expression reference (HPA tissue IHC: High in kidney tubule cells). Keep section thickness, cooling, antibody incubation, and detection conditions constant while assessing the retrieval step (standard IHC practice). If staining remains weak, adjust heating time in a small series and inspect tissue integrity before considering another retrieval buffer (standard IHC practice). The paraffin-section image for A05244-1 does not report its retrieval conditions (catalog antibody caption).
Could fixation explain weak or uneven RPLP2 staining?
Target-specific fixation sensitivity is unknown: the A05244-1 caption identifies paraffin-embedded prostate carcinoma tissue but does not state a fixative or fixation duration (catalog antibody caption). Compare sections with documented, consistent fixation and processing histories before changing antibody concentration (standard IHC practice). Patchy staining near poorly preserved areas can reflect processing variation, so check morphology and repeat the stain on an independently processed section (standard IHC practice). Do not assign a preferred fixative or fixation time to RPLP2 from its ribosomal function or modified residues (UniProt P05387: function and modified residues). Record fixation history with each scored slide to make differences interpretable (standard IHC practice).
What compartment should count as credible RPLP2 staining in tissue sections?
Expect predominantly cytoplasmic chromogenic staining in tissue, consistent with the reported general cytoplasmic IHC profile (HPA tissue IHC). Kidney tubule cells provide a high-expression comparison, whereas prostate glandular cells are reported at low expression despite the catalog’s prostate carcinoma image (HPA tissue IHC; catalog antibody caption). Nuclear speckles, cytosol, and mitotic spindle are reported from cellular localisation data, but their appearance in paraffin tissue requires separate assessment (HPA subcellular). Score cytoplasmic and nuclear staining separately, and compare each pattern with morphology and a negative control (standard IHC practice). Treat an isolated nuclear or extracellular deposit as inconclusive until independently reproduced (standard IHC practice).
Could epitope choice or modifications explain discordant RPLP2 staining?
The record lists one 115-aa chain and 0 annotated isoforms, so an isoform-specific tissue pattern has no support here (UniProt P05387). It lists phosphorylation at residues 17 and 19 and alternative modifications at residue 21, but supplies no antibody epitope or demonstrated effect on IHC binding (UniProt P05387; catalog antibody caption). The peptide-blocked prostate carcinoma image supports peptide-competitive staining for A05244-1, without identifying which residue is recognized (catalog antibody caption). If results conflict, compare the same sections with a second antibody whose epitope is documented, using matched controls (standard IHC practice). Avoid attributing a staining difference to a particular modification without direct validation (standard IHC practice).
How can I check RPLP2 localisation by multiplex immunofluorescence?
Pair RPLP2 with a validated marker for the expected cell population, such as kidney tubule cells, and assess overlap at the cell level (HPA tissue IHC: High in kidney tubule cells; standard IF practice). Choose spectrally separated fluorophores and place the weaker signal in a channel with low measured tissue autofluorescence (standard IF practice). Because RPLP2 has no transmembrane segment and is reported in cytosol, permeabilise to reach its intracellular epitope, then titrate permeabilisation against morphology and background (UniProt P05387 topology; HPA subcellular; standard IF practice). Include single-stain and no-primary controls to check bleed-through and background before interpreting nuclear speckles or spindle staining (HPA subcellular; standard IF practice).
What should I change when diffuse DAB obscures cytoplasmic RPLP2?
First compare the stained section with a no-primary control and inspect whether DAB appears in the same structures (standard IHC practice). Quench endogenous peroxidase, use an appropriate protein block, and wash thoroughly before adjusting antibody concentration or incubation time (standard chromogenic IHC practice). Titrate the catalog antibody across adjacent sections and retain the lowest concentration that preserves interpretable cytoplasmic staining in a positive reference (HPA tissue IHC: general cytoplasmic expression; standard IHC practice). Inspect tissue folds, cut edges, and necrotic regions separately because they can accumulate nonspecific signal (standard IHC practice). The peptide-blocked A05244-1 image is a specificity check, not a substitute for slide-level background controls (catalog antibody caption; standard IHC practice).
How should I quantify RPLP2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and regions of interest before scoring; kidney tubule cells and prostate glandular cells have different reported staining levels (HPA tissue IHC: High and Low, respectively; standard IHC practice). For cytoplasmic DAB, record the percentage of cells at each intensity and calculate an H-score from 0–300, or report percentage positive using a fixed threshold (standard IHC practice). Normalise positive-cell counts to eligible cells, or report positive-cell density per mm² of viable tissue when cell counts are impractical (standard IHC practice). Keep retrieval, detection, imaging, and threshold settings matched across groups, and exclude necrosis and section edges by predefined rules (standard IHC practice).
How can I distinguish true RPLP2 signal from staining artefacts?
A credible tissue result shows reproducible cytoplasmic staining in intact cells, consistent with the general tissue profile, and can be compared with high staining in kidney tubule cells (HPA tissue IHC). Interpret a negative area in context: adipocytes and skeletal myocytes are reported as not detected, while prostate glandular cells are reported at low levels (HPA tissue IHC). Suspect artefact when signal concentrates at cut edges, necrosis, folds, or structures positive in a no-primary control (standard IHC practice). Check endogenous peroxidase activity before attributing DAB deposits to RPLP2, and treat an isolated nuclear pattern cautiously because nuclear speckles were reported in subcellular imaging (HPA subcellular; standard IHC practice).
Boster reagents
Best RPLP2 / Large ribosomal subunit protein P2 IHC Antibodies
A05244-1 has a real IHC image from paraffin-embedded human prostate carcinoma tissue (catalog image caption); IHC, IF and ICC are listed for human, mouse and rat (catalog applications and reactivity).
Real IHC data
Anti-RPLP2/Ribosomal Protein Lp2 Antibody
Cat # A05244-1
A05244-1 will render with an IHC image of paraffin-embedded human prostate carcinoma tissue and a peptide-blocked comparison (catalog image caption). IF and ICC are listed applications, with human, mouse and rat reactivity, but no IF image is supplied (catalog applications, reactivity and image payload).
Which to pick: For tissue IHC, choose A05244-1: its image shows staining in a paraffin-embedded human prostate carcinoma section; the fixative is unreported (catalog image caption). For IF/ICC or work across human, mouse and rat, A05244-1 is the listed polyclonal option on the strength of its application and reactivity listings; no IF image is supplied (catalog dilution record, applications, reactivity and image payload).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P05387 (RLA2_HUMAN, Large ribosomal subunit protein P2).
- Human Protein Atlas. RPLP2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. RPLP2 subcellular location (ICC-IF): Mainly localized to the nuclear speckles and cytosol. In addition localized to the mitotic spindle..
- Human Protein Atlas. RPLP2 antibody validation summary (2 antibodies).
- Targeting RPLP2 Triggers DLBCL Ferroptosis by Decreasing FXN Expression. Biomedicines 2025 — PMC12189634.
- RPLP2 activates TLR4 in an autocrine manner and promotes HIF-1α-induced metabolic reprogramming in hepatocellular carcinoma. Cell death discovery 2023 — PMC10697958.
- Regulating the lncRNA DSCR9/RPLP2/PI3K/AKT axis: an important mechanism of Xinfeng capsules in improving rheumatoid arthritis. Frontiers in immunology 2024 — PMC11456487.
- Differentially expressed and survival-related proteins of lung adenocarcinoma with bone metastasis. Cancer medicine 2018 — PMC5911611.
- PubMed PMID:3323886 — UniProt-cited evidence.
- PubMed PMID:2153399 — UniProt-cited evidence.
- PubMed PMID:11875025 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RPLP2 (Large ribosomal subunit protein P2). Expected localisation General cytoplasmic staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A05244-1 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in tubules); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human prostate carcinoma tissue; fixative not specified (datasheet A05244-1); 2. Fixation: Image fixative and duration unreported (datasheet A05244-1); 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-RPLP2, 1:100 - 1:300 (datasheet A05244-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent (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 A05244-1); Caveat: Adipocytes and myocytes have no detected signal (HPA tissue IHC); Isoform / epitope: No isoforms; one 1–115 chain (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt P05387, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nuclear speckles and cytosol. In addition…, Datasheet A05244-1, PMC12189634, PMC10697958. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RPLP2/immunohistochemistry-ihc-guide.