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RPP30 / Ribonuclease P protein subunit p30 · IHC design guide
Design Immunohistochemistry for RPP30
Plan RPP30 paraffin IHC around nuclear staining that is most abundant in a subset of immune cells (HPA tissue IHC). Use bone marrow hematopoietic cells or spleen red-pulp cells as positive controls (HPA tissue IHC), and titrate the catalog antibody at 1:100–1:200 (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RPP30 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 | Nuclear staining in tissue sections (HPA tissue IHC) | |
| Staining pattern | Nuclear staining, strongest in a subset of immune cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Bone marrow+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Marrow and spleen peroxidase may mimic positive chromogen (HPA tissue IHC; standard IHC practice) | |
| Regulation | Specific expression regulator unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope impact unresolved (UniProt) |
Section 1
Recommended RPP30 IHC & IF Protocols
The catalog antibody’s IHC-P protocol and published RPP30 staining workflows in gastric tissue (PMC9343801) and mouse growth plates (PMC5527100) provide starting points for paraffin sections.
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A07553) |
| Fixation | Image fixative and duration unreported (datasheet A07553); 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 Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear 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-RPP30, 1:100-1:200 (datasheet A07553) |
| 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 | RPP30-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in several tissues, most abundant in a subset of immune cells. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0, 95–98 °C, 20 min for nuclear RPP30 (page retrieval rule; UniProt P78346); compare the published citrate conditions (PMC9343801; PMC5527100).
Section 2
What Is the Expected RPP30 Staining Pattern?
RPP30 is a nuclear protein associated with RNase P and RNase MRP (UniProt P78346); it has no transmembrane segment (UniProt P78346 topology). In paraffin-section IHC, expect nuclear staining across several tissues, strongest in subsets of immune cells (HPA tissue IHC). HPA rates the tissue staining Approved, with medium agreement with RNA data and external verification pending (HPA tissue IHC).
| Distinct nuclear stain in bone marrow hematopoietic cells or spleen red-pulp cells (HPA tissue IHC). | This matches the reported high signal in those cell populations (HPA tissue IHC). Assess nuclei within the named population, since HPA's levels describe cells within a tissue rather than every cell on the section (HPA tissue IHC). |
| Stain is predominantly cytoplasmic, membranous or extracellular, with little nuclear signal. | That conflicts with the nuclear tissue profile (HPA tissue IHC) and nuclear location (UniProt P78346). Consider artefact or nonspecific detection; a minor cytosolic signal alone is less decisive because HPA also reports cytosol in ICC-IF (HPA subcellular). |
| Strong stain appears in adipocytes or cardiomyocytes, while the expected nuclei remain unstained. | HPA reports RPP30 as not detected in those specific cell types (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity (general IHC practice); compare a known-positive section and an appropriate detection control before assigning target expression. |
| Broad, hazy chromogen covers cells and surrounding tissue without clear nuclear boundaries. | This does not resolve the expected nuclear pattern (HPA tissue IHC; UniProt P78346). Background from blocking, antibody concentration or detection chemistry can obscure localisation (general IHC practice); a darker field alone is insufficient evidence of RPP30. |
| No nuclear stain is visible in bone marrow hematopoietic cells on a candidate positive section. | HPA reports high staining in those cells (HPA tissue IHC), so review the run before interpreting a negative specimen. The absent signal does not identify which step failed; section quality, retrieval and detection require separate checks (general IHC practice). |
💡Expected RPP30 appearanceCall positive when a distinct nuclear signal is strongest in HPA-reported high-staining cell populations, such as bone marrow hematopoietic cells; widespread featureless stain or predominantly extranuclear signal is questionable (HPA tissue IHC; UniProt P78346).
| Which cells provide an IHC reference? (HPA tissue IHC) | Bone marrow hematopoietic cells, spleen red-pulp cells and testis Leydig cells are reported High; adrenal and breast glandular cells are Medium (HPA tissue IHC). Compare the named cell population, since neighbouring cells need not share its score (HPA tissue IHC). |
| How firm is the tissue-pattern evidence? (HPA tissue IHC) | HPA marks the tissue result Approved, yet reports medium staining-to-RNA consistency and pending external verification (HPA tissue IHC). The listed HPA037578 antibody is IHC Approved; the supplied record does not mark it Enhanced (HPA antibodies). |
| Can IF/ICC help interpret compartment? (HPA subcellular) | Yes. HPA approves nucleoplasm and nucleoli as main ICC-IF locations, with cytosol and microtubule ends as additional locations (HPA subcellular). Use that as a localisation cross-check; the IHC tissue profile remains nuclear (HPA tissue IHC). |
| Do isoforms or processing specify a different IHC pattern? (UniProt P78346) | UniProt lists 2 isoforms and a chain spanning residues 2–268, with no signal peptide or propeptide (UniProt P78346). No antibody epitope or isoform-specific staining evidence is supplied, so these annotations cannot predict separate staining patterns. |
| Is RPP30 fixation sensitivity established? (supplied UniProt and HPA records) | No target-specific fixation effect is reported in the supplied records (UniProt P78346; HPA tissue IHC). Use the catalog antibody's validated IHC-P conditions as the starting point; assess retrieval performance with controls (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive nuclei are blank. | A failed staining step or unsuitable section is possible (general IHC practice); high bone marrow hematopoietic staining is reported (HPA tissue IHC). | Check the catalog antibody's IHC-P conditions, retrieval, reagent activity and positive-section integrity one step at a time (general IHC practice). |
| Chromogen fills cytoplasm but leaves nuclei pale. | The pattern conflicts with nuclear tissue staining (HPA tissue IHC); cytosol is only an additional ICC-IF location (HPA subcellular). | Review morphology and counterstain, then compare an HPA-reported positive cell population and detection controls (HPA tissue IHC; general IHC practice). |
| Adipocytes, cardiomyocytes or liver cholangiocytes stain strongly. | These named cell types are reported Not detected (HPA tissue IHC); nonspecific binding or endogenous detection activity is possible (general IHC practice). | Confirm cell identity and examine a no-primary detection control before interpreting the stain as RPP30 (general IHC practice). |
| The whole section has a uniform brown haze. | Nuclear localisation is obscured (HPA tissue IHC); excessive antibody or detection background may contribute (general IHC practice). | Review blocking, washing and the catalog antibody's IHC-P dilution guidance; compare the no-primary control (general IHC practice). |
| A presumed negative tissue has scattered nuclear-positive cells. | HPA's Not detected calls apply to specified cell types, not necessarily every cell in that tissue (HPA tissue IHC). | Identify the stained cells before scoring. Compare their compartment and intensity with an HPA-reported positive population (HPA tissue IHC; general IHC practice). |
| Replicate sections yield inconsistent nuclear intensity. | Section or run variation is possible (general IHC practice); HPA reports only medium agreement between staining and RNA data (HPA tissue IHC). | Run comparable positive sections together, hold detection conditions constant and score the same cell population (general IHC practice). |
Experiment evidence
IHC & IF images showing RPP30 staining
A07553 · IHCImmunohistochemistry of paraffin-embedded human lung cancer using RPP30 antibody at dilution of 1:100 .Perform microwave antigen retrieval w
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000148688-RPP30/tissue/cerebral+cortex#imid_8988050
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000148688-RPP30/tissue/colon#imid_8985368
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000148688-RPP30/tissue/liver#imid_8985369
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000148688-RPP30/tissue/kidney#imid_8985378
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037578; green = RPP30, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037578; green = RPP30, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RPP30 IHC & IF
🧪Run bone marrow first: hematopoietic cells should stain (HPA: High in bone marrow hematopoietic cells). Use adipose tissue as the negative tissue: adipocytes should lack detectable staining (HPA: Not detected in adipose tissue adipocytes); on the positive slide, cells without target signal should show counterstain only, although the supplied HPA rows do not identify a specific internal negative bone marrow cell type.
Positive control tissue: Bone marrow (Hematopoietic cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RPP30 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved), Nucleoli (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and clonality, and a biological negative such as RPP30 knockout material or a peptide block if the immunizing peptide is available (standard IHC practice). Quench endogenous peroxidase in bone marrow for chromogenic detection, and assess autofluorescence if using fluorescence detection (standard IHC/IF practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence; the selected A07553 paraffin-section caption does not state a fixative (A07553 tissue-IHC caption). That caption uses microwave retrieval in 10 mM PBS, pH 7.2, at 1:100 dilution in lung cancer, but does not establish that retrieval is required for other tissues or that frozen sections or IF are easier (A07553 tissue-IHC caption). Bone marrow may have endogenous peroxidase background during chromogenic detection, so interpret staining against the no-primary control (standard IHC practice).
HPA tissue IHC evidence for RPP30
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | High | Protein (IHC) | HPA → |
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RPP30 IHC Tips
Troubleshoot RPP30 staining in paraffin sections by checking retrieval, nuclear localisation, controls, and cell-specific scoring before interpreting chromogenic signal.
How should I retrieve RPP30 when nuclear staining is weak?
Start with Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval rule: nuclear antigen). Cool sections consistently, then compare nuclear staining and tissue integrity against a matched control slide (standard IHC practice). If staining remains weak, test microwave retrieval in 10 mM PBS at pH 7.2, the condition reported for catalog antibody A07553 at 1:100 in paraffin-embedded human lung cancer (A07553 tissue-IHC caption). Change one retrieval variable at a time; overly harsh heating can impair morphology and complicate scoring (standard IHC practice).
Can fixation explain absent RPP30 staining in paraffin sections?
RPP30-specific fixation sensitivity is unknown because the supplied product caption does not state a fixative (A07553 tissue-IHC caption). Record the actual fixative, fixation duration, processing history, and section age for each specimen before attributing a weak result to fixation (standard IHC practice). Compare sections processed together using the same retrieval and detection conditions, with a suitable positive control on each run (standard IHC practice). If staining differs, repeat with matched material and adjust retrieval systematically; tissue staining patterns and RPP30 protein annotations cannot establish a fixation effect (HPA tissue IHC; UniProt P78346).
What RPP30 staining pattern should I expect in tissue?
Prioritise nuclear staining: RPP30 is annotated in the nucleus and nucleolus (UniProt P78346), and tissue IHC reports nuclear expression across several tissues (HPA tissue IHC). Nucleoplasmic and nucleolar signals are approved in cell imaging, with additional cytosolic and microtubule-end locations (HPA subcellular). Score nuclear signal separately from cytoplasmic signal, using the same counterstain and compartment criteria across slides (standard IHC practice). If only diffuse cytoplasmic chromogen appears, review antibody concentration, background, and nuclear preservation before assigning that pattern to RPP30 (HPA subcellular; standard IHC practice).
Could an isoform or modified epitope alter RPP30 IHC staining?
RPP30 has 2 listed isoforms, but the payload does not map this antibody’s epitope to either isoform (UniProt P78346; A07553 tissue-IHC caption). Its annotated modifications include N-acetylalanine at residue 2 and phosphoserine at residue 251; their effect on this antibody is unknown (UniProt P78346). Check the antibody’s documented immunogen or epitope information before claiming isoform selectivity, and compare retrieval conditions on matched sections if epitope exposure is suspected (standard IHC practice). Keep an observed staining difference separate from a proposed molecular cause until an independent assay supports that cause (standard IHC practice).
How can IF help resolve an ambiguous chromogenic RPP30 pattern?
Use IF as a separate validation experiment and compare its nuclear pattern with the paraffin-section IHC result (UniProt P78346; HPA subcellular). Multiplex RPP30 with a marker for the cell type being evaluated, and include single-label controls to check channel bleed-through (standard IF practice). Choose a fluorophore and filter combination away from the specimen’s strongest autofluorescence, then inspect unstained tissue at identical exposure settings (standard IF practice). Because RPP30 lacks a transmembrane segment and is primarily nuclear, permeabilise sufficiently for antibody access to nuclear epitopes while checking that nuclear morphology remains intact (UniProt P78346 topology; standard IF practice).
How do I troubleshoot diffuse or granular RPP30 chromogen?
Compare stained sections with a no-primary control and inspect whether signal follows nuclei or accumulates in tissue deposits and damaged areas (standard IHC practice; UniProt P78346 localisation). Optimise primary antibody concentration from the reported 1:100 starting condition for A07553, while holding retrieval and detection constant (A07553 tissue-IHC caption; standard IHC practice). For peroxidase-based detection, verify the endogenous peroxidase block and shorten chromogen development if background obscures nuclei (standard IHC practice). Check that blocking, washes, and counterstaining are consistent across runs before treating a diffuse pattern as biological RPP30 expression (standard IHC practice).
How should I quantify RPP30 across heterogeneous tissue? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score nuclear chromogen separately from any cytoplasmic signal (UniProt P78346 localisation; standard IHC practice). For that population, report the percentage of positive nuclei and, where intensity is reproducible, an H-score using 0–3 intensity grades (standard IHC practice). Normalise positive counts to the total evaluable cells of the same type, or report positive-cell density per mm² of evaluable tissue (standard IHC practice). Apply one threshold, comparable section quality, and the same exclusion rules for folds and necrosis across specimens (standard IHC practice).
When is a positive RPP30 IHC result convincing?
A convincing result has reproducible nuclear staining in intact cells, consistent with RPP30’s nuclear and nucleolar annotation (UniProt P78346; HPA tissue IHC). Interpret the stained cell type explicitly: HPA reports high signal in bone-marrow hematopoietic cells and spleen red-pulp cells, while lung macrophages are listed as low (HPA tissue IHC). Treat staining confined to section edges, necrotic regions, or structures with endogenous enzyme activity as suspect until controls resolve it (standard IHC practice). The HPA tissue-IHC assessment is Approved with medium staining-versus-RNA consistency and pending external verification, so corroborate consequential findings independently (HPA tissue IHC).
Boster reagents
Best RPP30 / Ribonuclease P protein subunit p30 IHC Antibodies
A07553 has a real paraffin-section IHC image from human lung cancer (catalog image caption) and lists IF/ICC use and Human, Mouse, and Rat reactivity (catalog applications and reactivity).
Real IHC data
Anti-RPP30 Antibody
Cat # A07553
A07553 will render with an IHC image of paraffin-embedded human lung cancer stained at 1:100 after microwave retrieval in 10 mM PBS, pH 7.2 (catalog image caption). The catalog lists A07553 for IHC and IF/ICC in Human, Mouse, and Rat; it provides no IF image (catalog applications, reactivity, and image records).
Which to pick: For paraffin-section IHC, choose A07553 because its own image caption documents that preparation and staining conditions (catalog image caption); the fixative is unreported (catalog image caption). For IF/ICC or work across Human, Mouse, and Rat, A07553 is the listed option (catalog applications and reactivity), with IF/ICC at 1:50–1:200 (catalog dilution); its clone designation is unreported (catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P78346 (RPP30_HUMAN, Ribonuclease P protein subunit p30).
- Human Protein Atlas. RPP30 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. RPP30 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and nucleoli. In addition localized to the cytosol and microtubule ends..
- Human Protein Atlas. RPP30 antibody validation summary (1 antibodies).
- RPP30 is a novel diagnostic and prognostic biomarker for gastric cancer. Frontiers in genetics 2022 — PMC9343801.
- Expression of RMRP RNA is regulated in chondrocyte hypertrophy and determines chondrogenic differentiation. Scientific reports 2017 — PMC5527100.
- SARS-CoV-2 RNA detection in Formalin-Fixed Paraffin-Embedded (FFPE) tissue by droplet digital PCR (ddPCR). Clinica chimica acta; international journal of clinical chemistry 2022 — PMC9085371.
- PubMed PMID:9037013 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15164054 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RPP30 (Ribonuclease P protein subunit p30). Expected localisation Nuclear staining in tissue sections (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A07553 at 1:100-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A07553); 2. Fixation: Image fixative and duration unreported (datasheet A07553); 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 Tris-EDTA buffer, pH 9.0 , 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-RPP30, 1:100-1:200 (datasheet A07553); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Marrow and spleen peroxidase may mimic positive chromogen (HPA tissue IHC; standard IHC practice); Isoform / epitope: 2 isoforms; epitope impact unresolved (UniProt); Regulation: Specific expression regulator unreported (UniProt). Evidence: UniProt P78346, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm and nucleoli. In addition…, Datasheet A07553, PMC9343801, PMC5527100. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RPP30/immunohistochemistry-ihc-guide.