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MRPS5 / Small ribosomal subunit protein uS5m · IHC design guide
Design Immunohistochemistry for MRPS5
Plan MRPS5 paraffin-section IHC around cytoplasmic tissue staining (HPA tissue IHC) and mitochondrial localization (UniProt). Start with the catalog antibody’s IHC dilution range of 1:100–1:300 (datasheet), using kidney tubule cells as a high-staining reference and adipocytes as an undetected reference (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MRPS5 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 tissue staining (HPA tissue IHC); mitochondrial protein (UniProt) | |
| Staining pattern | Widespread cytoplasmic staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Breast+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 sections (standard IHC practice; not target-specific) | |
| Caveat | Staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | No specific regulator annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unreported (UniProt) |
Section 1
Recommended MRPS5 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with a published human clear cell renal cell carcinoma protocol (PMC11614452: immunohistochemical staining).
| Sample | Paraffin-embedded Human testis tissue; fixative not specified (datasheet A14183S5) |
| Fixation | Image fixative and duration unreported (datasheet A14183S5); 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-MRPS5, 1:100-1:300 (datasheet A14183S5) |
| 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 | MRPS5-positive staining in myoepithelial cells of breast (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule); the published protocol used microwave EDTA retrieval (PMC11614452: methods).
Section 2
What Is the Expected MRPS5 Staining Pattern?
MRPS5 is a mitochondrial small ribosomal subunit protein with no transmembrane segment (UniProt P82675). In paraffin tissue sections, expect predominantly cytoplasmic staining, including strong staining in kidney tubular cells and duodenal glandular cells (HPA: tissue IHC). HPA rates its IHC antibody Approved, but reports low consistency with RNA expression and pending external verification (HPA: reliability).
| Cytoplasmic staining is strongest in kidney tubular cells or duodenal glandular cells (HPA: High in both). | This fits the reported tissue pattern (HPA: tissue IHC). A punctate cytoplasmic appearance would be compatible with mitochondrial localization, but chromogenic IHC alone cannot confirm organelle identity (UniProt P82675; HPA: mitochondria supported in ICC-IF). |
| A strong, predominantly nuclear or membrane-rim signal replaces cytoplasmic staining (HPA: ubiquitous cytoplasmic profile). | That compartment is unexpected for MRPS5 (UniProt P82675: mitochondrion; no transmembrane segment). Check morphology and detection controls before scoring it as specific; aberrant staining can reflect an IHC artefact (general IHC practice). |
| Strong staining appears in adipocytes or liver cholangiocytes (HPA: Not detected in those cells). | Treat the result as discordant with HPA's cell-level observations, rather than proof that the whole tissue is negative. Cross-reactivity or endogenous detection activity is possible; compare a no-primary control (HPA: tissue IHC; general IHC practice). |
| Colour spreads across tissue and blank areas without a discernible cell pattern (general IHC practice). | Diffuse background prevents a reliable MRPS5 call. Review blocking, washes and detection controls before interpreting intensity (general IHC practice); HPA reports cell-associated cytoplasmic staining (HPA: tissue IHC). |
| No signal appears in kidney tubular cells despite intact tissue morphology (HPA: High in kidney tubular cells). | A failed run is plausible, but a negative result alone cannot identify its cause. Check the antibody and detection steps, then review retrieval conditions as a general IHC troubleshooting step (HPA: tissue IHC; general IHC practice). |
💡Expected MRPS5 appearanceCall a convincing positive when kidney tubular or duodenal glandular cells show strong, predominantly cytoplasmic staining (HPA: High in both; ubiquitous cytoplasmic profile); isolated dominant nuclear, membrane-rim or diffuse background staining is discordant (UniProt P82675: mitochondrion; general IHC practice).
| Which cells provide useful comparisons? (HPA: tissue IHC) | Kidney tubular and small-intestinal glandular cells are High; adipocytes are Not detected (HPA: tissue IHC). Score the named cells, since HPA levels do not describe every cell in a section. |
| How firm is the tissue pattern? (HPA: reliability) | The IHC antibody is Approved, with low consistency between staining and RNA expression and external verification pending (HPA: reliability); use discordant staining as a reason to investigate. |
| Do isoforms or processing change the expected slide pattern? (UniProt P82675) | Two isoforms are listed, and the annotated chain spans residues 1–430 (UniProt P82675). Epitope coverage by isoform is unspecified, so no isoform-specific IHC pattern can be predicted. |
| Does protein topology suggest a surface or shed signal? (UniProt P82675) | MRPS5 has no annotated transmembrane segment, signal peptide or glycosylation sites (UniProt P82675); these annotations offer no basis for expecting membrane-rim staining or extracellular deposition. |
| IF/ICC: where should signal localize? (HPA: subcellular ICC-IF) | Mainly mitochondria; additional midbody and primary-cilium locations are uncertain (HPA: subcellular ICC-IF). This is an IF localization answer, not a paraffin IHC protocol or proof of those structures in tissue. |
| Situation | Likely cause | Next action |
|---|---|---|
| Kidney tubular cells are unstained (HPA: High in kidney tubular cells). | Primary antibody, retrieval or detection may have failed (general IHC practice); HPA provides no MRPS5-specific retrieval requirement. | Review the validated IHC conditions and detection controls, then adjust retrieval only as a general IHC optimization (general IHC practice). |
| Nuclei dominate while cytoplasm is faint (HPA: ubiquitous cytoplasmic profile). | The pattern conflicts with mitochondrial localization (UniProt P82675); nonspecific signal or interpretation error is possible (general IHC practice). | Recheck cell boundaries and the no-primary control; score MRPS5 only where a credible cytoplasmic pattern remains (general IHC practice; HPA: tissue IHC). |
| Adipocytes show strong stain (HPA: Not detected in adipocytes). | Cross-reactivity or endogenous detection activity may contribute (general IHC practice); HPA's negative call is cell-specific (HPA: tissue IHC). | Compare no-primary and detection controls, and confirm that the scored cells are adipocytes (general IHC practice; HPA: tissue IHC). |
| Diffuse colour obscures glandular cells (general IHC practice). | Excess background can arise during antibody incubation or detection (general IHC practice); it cannot establish MRPS5 distribution. | Check blocking, antibody concentration and washes using the validated IHC workflow, then reassess cellular contrast (general IHC practice). |
| A negative tissue shows some positive cells (HPA: cell-level tissue IHC). | HPA's Not detected calls name particular cells, such as liver cholangiocytes, rather than every cell in that tissue (HPA: tissue IHC). | Identify the stained cell type before calling the result discordant; compare the matching HPA cell-level entry (HPA: tissue IHC). |
| The result changes with fixation or retrieval (general IHC practice). | The supplied record does not establish MRPS5-specific fixation sensitivity or epitope masking (UniProt P82675; HPA: tissue IHC). | Document processing conditions and compare matched controls; avoid assigning a target-specific mechanism from this change alone (general IHC practice). |
Experiment evidence
IHC & IF images showing MRPS5 staining
A14183S5 · IHCImmunohistochemistry validation of MRPS5 using Anti-MRP-S5 Antibody (A14183S5). Immunohistochemical analysis of paraffin-embedded Human test
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000144029-MRPS5/tissue/cerebral+cortex#imid_15584839
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000144029-MRPS5/tissue/colon#imid_15584765
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000144029-MRPS5/tissue/liver#imid_15584758
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000144029-MRPS5/tissue/kidney#imid_15584751
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA055765; green = MRPS5, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA055765; green = MRPS5, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MRPS5 IHC & IF
🧪Run kidney first: tubular cells should stain strongly (HPA: High in kidney cells in tubules). Use adipose tissue as the negative comparator (HPA: Not detected in adipocytes); on the kidney slide, non-tubular cells without staining can serve as candidate internal background controls, but their MRPS5-negative status is not established by the supplied HPA row.
Positive control tissue: Breast (Myoepithelial cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MRPS5 in A-549, U2OS, KOLF2.1J, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; use a host- and class-matched isotype control for a monoclonal primary or matched nonimmune IgG for a polyclonal primary; and compare with MRPS5-knockout material processed identically (standard IHC practice). For kidney chromogenic IHC, block endogenous peroxidase and, if using avidin–biotin detection, check for endogenous biotin background (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A14183S5 paraffin-section caption does not report a fixative (selected tissue-IHC caption). Antigen retrieval dependence is unreported, so retrieval conditions need empirical optimization; the supplied evidence does not establish that frozen sections or IF are easier (selected tissue-IHC caption; HPA: supported mitochondrial ICC-IF localization). In kidney, endogenous biotin can complicate avidin–biotin detection (standard IHC practice).
HPA tissue IHC evidence for MRPS5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Breast | Myoepithelial cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Small intestine | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MRPS5 IHC Tips
Troubleshoot MRPS5 chromogenic IHC in paraffin sections by checking retrieval, processing, cytoplasmic localisation and cell-specific controls before interpreting staining intensity.
What retrieval should I start with for MRPS5 paraffin-section IHC?
Start with citrate buffer at pH 6.0, heat-induced retrieval at 95–98 °C for 20 min (page retrieval rule). Process control and test sections together, cool them after heating, and keep chromogenic detection conditions identical when comparing retrieval outcomes (standard IHC practice). MRPS5 is mitochondrial and has no transmembrane segment, but the antibody epitope and its retrieval sensitivity are unspecified (UniProt P82675 localisation and topology; supplied antibody evidence). If staining remains weak, test another retrieval condition only as a documented fallback on serial sections, assessing signal alongside tissue morphology and background (standard IHC practice).
How should I assess fixation when MRPS5 IHC staining varies between blocks?
Record the fixative, fixation duration and processing for each block; the A14183S5 image caption identifies paraffin-embedded human testis but does not report its fixative (selected tissue-IHC caption). Target-specific MRPS5 fixation sensitivity is unknown, and neither its tissue staining pattern nor its topology or modification record establishes an optimal fixation schedule (supplied evidence; UniProt P82675). Compare consistently processed sections with a staining control in each run, keeping detection conditions constant (standard IHC practice). When assessing a fixation difference, hold citrate retrieval at pH 6.0 for 20 min so retrieval does not change at the same time (page retrieval rule; standard IHC practice).
Where should convincing MRPS5 staining appear in a chromogenic section?
Expect predominantly cytoplasmic staining consistent with mitochondrial localisation, rather than treating nuclear staining alone as convincing MRPS5 signal (HPA tissue IHC profile; HPA subcellular: mitochondria supported; UniProt P82675 localisation). The HPA profile describes ubiquitous cytoplasmic expression, while its subcellular data also list midbody and primary cilium localisation as uncertain (HPA tissue IHC; HPA subcellular). Inspect staining at high magnification within intact cells and compare its distribution with adjacent background and a matched control section (standard IHC practice). Score the cytoplasmic compartment consistently; a sharply nuclear or extracellular pattern warrants checks of retrieval, blocking and detection before biological interpretation (standard IHC practice).
Can this stain distinguish MRPS5 isoforms or reveal epitope loss?
MRPS5 has 2 annotated isoforms and an S5 DRBM region at residues 218–282, but the supplied antibody evidence does not locate its epitope (UniProt P82675; supplied antibody evidence). Consequently, chromogenic staining cannot be assigned to one isoform from this evidence, even when cell-level patterns differ (UniProt P82675 isoforms; supplied antibody evidence). The record lists no glycosylation sites or modified residues, which does not establish how fixation or retrieval affects antibody binding (UniProt P82675; supplied fixation evidence). If an isoform-specific conclusion matters, obtain documented epitope coverage and use an independently validated isoform assay alongside matched IHC controls (standard assay-validation practice).
How can IF help assess an ambiguous MRPS5 IHC pattern?
Use IF as a separate localisation check when chromogenic cytoplasmic staining is ambiguous; mitochondrial localisation is supported, while reported midbody and primary cilium localisation remains uncertain (HPA subcellular). Multiplex an MRPS5 channel with a mitochondrial marker and a marker for the expected cell type, such as kidney tubular cells, and compare signals within the same cells (HPA subcellular; HPA tissue IHC: high in kidney tubules; standard IF practice). Choose a far-red fluorophore where tissue autofluorescence compromises shorter wavelengths, with single-channel controls (standard IF practice). Because the antibody epitope’s side of a mitochondrial membrane is unspecified, titrate permeabilisation rather than assuming access; a mild 0.1% detergent treatment for 5–10 min is a starting condition, not MRPS5-specific validation (UniProt P82675 topology; supplied antibody evidence; standard IF practice).
How do I separate MRPS5 signal from chromogenic background?
Inspect a no-primary control alongside the stained section to identify secondary-reagent or detection background (standard IHC practice). In a peroxidase-based workflow, an endogenous peroxidase block, such as 3% hydrogen peroxide for 10 min, and DAB development are general IHC steps, not MRPS5-specific evidence (standard IHC practice). Keep blocking, antibody incubation, washes and DAB development comparable across sections before attributing a diffuse brown haze to MRPS5 (standard IHC practice). If background persists, examine section edges, damaged areas and extracellular deposits separately from intact-cell cytoplasm; the expected protein location is mitochondrial (standard IHC practice; UniProt P82675 localisation).
How should I quantify MRPS5 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic scoring compartment before reading slides, because the reported tissue profile is cytoplasmic (HPA tissue IHC profile; standard IHC practice). For an H-score, grade viable cells from 0–3, multiply each grade by its percentage of cells, and sum to a 0–300 score (standard IHC scoring practice). Alternatively, report the percentage of positive cells or positive-cell density per mm², normalising the latter to viable tissue area rather than the entire section (standard IHC quantification practice). Apply one threshold, control and sampling rule across cases, and record cell type because HPA reports different staining levels among cell populations (HPA tissue IHC; standard IHC practice).
What makes an MRPS5-positive IHC result biologically credible?
Look for staining within intact-cell cytoplasm, consistent with MRPS5 mitochondrial localisation and the reported cytoplasmic tissue pattern (UniProt P82675 localisation; HPA tissue IHC profile). Cell identity matters: HPA reports high staining in kidney tubular cells but no detection in adipocytes, while warning of low consistency between staining and RNA expression data (HPA tissue IHC). An isolated nuclear signal, section-edge accentuation or staining confined to necrotic areas should prompt review of morphology and controls before calling a positive result (standard IHC practice). Compare a no-primary section and peroxidase-blocked detection run to assess reagent background or endogenous enzyme activity in chromogenic IHC (standard IHC practice).
Boster reagents
Best MRPS5 / Small ribosomal subunit protein uS5m IHC Antibodies
A14183S5 has real IHC data from human paraffin-embedded testis (catalog IHC caption). IF is listed for human and mouse, but no IF figure is supplied (catalog applications, reactivity and IF image alts).
Real IHC data
Anti-MRP-S5 Antibody
Cat # A14183S5
A14183S5 will render with its own IHC figure showing human paraffin-embedded testis at 1:100 (catalog IHC caption). The catalog also lists IF and human and mouse reactivity, without an IF figure or mouse tissue image (catalog applications, reactivity and image alts).
Which to pick: For tissue IHC, choose A14183S5: its own image shows human paraffin-embedded testis, and the fixative is unreported (catalog IHC caption). For IF/ICC, A14183S5 lists IF at 1:50, but ICC validation and an IF figure are unreported (catalog applications, IF dilution and IF image alts). For cross-species work, A14183S5 lists human and mouse reactivity; it has a rabbit host and unreported clonality, while its IHC image documents human tissue only (catalog reactivity, host and clone; catalog IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P82675 (RT05_HUMAN, Small ribosomal subunit protein uS5m).
- Human Protein Atlas. MRPS5 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MRPS5 subcellular location (ICC-IF): Mainly localized to the mitochondria. In addition localized to the midbody and primary cilium..
- Human Protein Atlas. MRPS5 antibody validation summary (1 antibodies).
- Low Expression of Mitochondrial Ribosomal Protein S5 is Associated With Poor Prognosis in Patients With Clear Cell Renal Cell Carcinoma. Applied immunohistochemistry & molecular morphology : AIMM 2025 — PMC11614452.
- Mutant MRPS5 affects mitoribosomal accuracy and confers stress-related behavioral alterations. EMBO reports 2018 — PMC6216279.
- PubMed PMID:11402041 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MRPS5 (Small ribosomal subunit protein uS5m). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); mitochondrial protein (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A14183S5 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Myoepithelial cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded Human testis tissue; fixative not specified (datasheet A14183S5); 2. Fixation: Image fixative and duration unreported (datasheet A14183S5); 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-MRPS5, 1:100-1:300 (datasheet A14183S5); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences unreported (UniProt); Regulation: No specific regulator annotated (UniProt). Evidence: UniProt P82675, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the mitochondria. In addition localized to the…, Datasheet A14183S5, PMC11614452. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MRPS5/immunohistochemistry-ihc-guide.