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MRPL58 / Large ribosomal subunit protein mL62 · IHC design guide
Design Immunohistochemistry for MRPL58
Plan chromogenic paraffin-section IHC for MRPL58 using the IHC-validated antibody at 1:25 (datasheet: A31846-1). Assess cytoplasmic tissue staining (HPA tissue IHC) while recognizing its mitochondrial localization (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MRPL58 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 | Observed cytoplasm (HPA tissue IHC); mitochondrial protein (UniProt) | |
| Staining pattern | Cytoplasmic staining in several tissues (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Bone marrow+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section 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 A31846-1) | |
| Caveat | Staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Down-regulated during HT29-D4 differentiation (UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain spans 30–206 (UniProt) |
Section 1
Recommended MRPL58 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet) with the published human osteosarcoma tissue method (PMC12450695 methods).
| Sample | Paraffin-embedded H. colorectal carcinoma tissue; fixative not specified (datasheet A31846-1) |
| Fixation | Image fixative and duration unreported (datasheet A31846-1); 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-MRPL58, 1:25 (datasheet A31846-1) |
| 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 | MRPL58-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in several tissues. 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 method does not specify retrieval conditions (PMC12450695 methods).
Section 2
What Is the Expected MRPL58 Staining Pattern?
MRPL58 is a mitochondrial protein with no transmembrane segment (UniProt Q14197). In paraffin section IHC, expect cytoplasmic staining in selected cells, including colon glandular cells and cerebral cortex neurons, where HPA reports medium staining (HPA tissue IHC). HPA rates its IHC evidence “Approved” but reports low consistency between antibody staining and RNA expression; interpret tissue differences with that limitation (HPA tissue IHC).
| Cytoplasmic staining in colon glandular cells or cerebral cortex neurons. | This matches the cells and medium levels reported by HPA (HPA tissue IHC). A fine granular appearance can fit a mitochondrial protein, but paraffin section IHC alone does not establish that individual deposits are mitochondria (UniProt Q14197; general IHC practice). |
| Predominantly nuclear or cell-border staining, with little cytoplasmic signal. | Treat this as a compartment mismatch for tissue IHC: HPA describes cytoplasmic tissue expression, while UniProt places MRPL58 in mitochondria (HPA tissue IHC; UniProt Q14197). Review controls and detection before calling it MRPL58; HPA's additional nucleoplasmic and plasma-membrane IF locations are uncertain (HPA subcellular). |
| Strong staining in a cell population HPA lists as “Not detected,” such as bone marrow hematopoietic cells. | This conflicts with that reported tissue result and warrants checks for cross-reactivity or endogenous detection activity (HPA tissue IHC; general IHC practice). “Not detected” describes HPA's assay observation, not proof that every specimen or method must be negative (HPA tissue IHC). |
| Uniform color across tissue and empty spaces, with poor cell boundaries. | Interpret this as background rather than a resolved cellular pattern (general IHC practice). Check the no-primary control, blocking, washes and chromogen development; HPA's reported MRPL58 pattern is cytoplasmic expression in several tissues (HPA tissue IHC). |
| No staining in colon glandular cells in a run intended to show a positive result. | HPA reports medium staining there, so review tissue preservation, retrieval, primary-antibody application and detection controls before interpreting the absence biologically (HPA tissue IHC; general IHC practice). HPA's low staining-to-RNA consistency also limits certainty from a single specimen (HPA tissue IHC). |
💡Expected MRPL58 appearanceCall a section positive when the relevant cells show discernible cytoplasmic staining, typically around the medium level reported for colon glands or cerebral cortex neurons; dominant nuclear, cell-border or cell-free color calls for control review (HPA tissue IHC; UniProt Q14197; general IHC practice).
| Cellular compartment | MRPL58 belongs to the mitochondrial large ribosomal subunit and has no transmembrane segment (UniProt Q14197). HPA describes tissue IHC as cytoplasmic; do not require a distinct mitochondrial outline in chromogenic sections (HPA tissue IHC; general IHC practice). |
| Tissue and cell choice | HPA reports medium staining in colon glandular cells and adipocytes, but “Not detected” in salivary glandular cells and skeletal myocytes (HPA tissue IHC). Compare the named cell populations, since a whole-section average can obscure their different reported results (general IHC practice). |
| Evidence strength | The tissue IHC reliability label is “Approved,” with low consistency between staining and RNA expression; the listed IHC antibody is also “Approved,” not “Enhanced” (HPA tissue IHC; HPA antibodies). Use the pattern as a reference for interpretation, with controls for each run (general IHC practice). |
| IF/ICC Q&A: Should nuclear or membrane fluorescence be expected? | Mitochondria are the supported main location; nucleoplasm and plasma membrane are additional uncertain locations (HPA subcellular). Treat those extra signals cautiously. This IF/ICC observation does not establish an IHC-P staining pattern (HPA subcellular; HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected cytoplasmic signal is absent in colon glands. | A staining-run failure is possible; HPA reports medium staining in these cells (HPA tissue IHC; general IHC practice). | Check the run's positive control, primary-antibody application, retrieval and detection steps, then repeat if a control failed (general IHC practice). No MRPL58-specific retrieval condition is established by the supplied evidence. |
| Color is strongest in nuclei or along cell borders. | The distribution conflicts with HPA's cytoplasmic tissue profile and UniProt's mitochondrial location (HPA tissue IHC; UniProt Q14197). | Compare with a no-primary control and reassess antibody specificity and detection background before assigning the signal to MRPL58 (general IHC practice). |
| Bone marrow hematopoietic cells stain strongly. | HPA lists these cells as “Not detected”; cross-reactivity or endogenous detection activity may explain unexpected color (HPA tissue IHC; general IHC practice). | Inspect the no-primary control and the detection and blocking steps; report the discrepancy by cell type rather than calling the specimen globally positive (general IHC practice). |
| Diffuse chromogen obscures cell boundaries. | Background from the staining workflow may overwhelm a cellular signal (general IHC practice). | Check blocking, wash quality and chromogen development against the no-primary control; score MRPL58 only where a cellular cytoplasmic pattern remains distinguishable (general IHC practice; HPA tissue IHC). |
| A reportedly negative cell type shows faint color. | “Not detected” is an HPA observation, and faint color alone cannot separate weak signal from background (HPA tissue IHC; general IHC practice). | Compare intensity with nearby background and run controls, document the named cell type, and avoid converting the HPA observation into an absolute biological negative (HPA tissue IHC; general IHC practice). |
| The slide pattern differs from a mitochondrial IF image. | HPA's supported mitochondrial localization comes from ICC-IF, while its tissue IHC profile is reported as cytoplasmic (HPA subcellular; HPA tissue IHC). | Judge chromogenic sections by cell-resolved cytoplasmic staining and IHC controls; use IF localization as context rather than requiring identical visual detail (HPA tissue IHC; HPA subcellular; general IHC practice). |
Experiment evidence
IHC & IF images showing MRPL58 staining
A31846-1 · IHCImmunohistochemical analysis of paraffin-embedded H. colorectal carcinoma section using ICT1 Antibody (C-term). A31846-1 was diluted at 1:25
A31846-1 · IHCImmunohistochemical analysis of paraffin-embedded H. skin section using ICT1 Antibody (C-term). A31846-1 was diluted at 1:25 dilution. A und
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000167862-MRPL58/tissue/cerebral+cortex#imid_1310124
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000167862-MRPL58/tissue/colon#imid_1310050
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000167862-MRPL58/tissue/liver#imid_1310051
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000167862-MRPL58/tissue/kidney#imid_1310068
HPA · ICC-IF OE19Human Protein Atlas immunofluorescence, OE19, antibody HPA076987; green = MRPL58, blue = nucleus, red = microtubules.
HPA · ICC-IF OE19Human Protein Atlas immunofluorescence, OE19, antibody HPA076987; green = MRPL58, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MRPL58 IHC & IF
🧪Start with adipose tissue: adipocytes should stain at a medium level (HPA: adipose tissue, adipocytes, Medium). Use bone marrow hematopoietic cells as the negative tissue (HPA: bone marrow, hematopoietic cells, Not detected); on the adipose slide, compare staining with adjacent non-adipocyte cells as an internal background reference, without assuming those cells are MRPL58-negative.
Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)
Negative control tissue: Bone marrow (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MRPL58 in OE19, PC-3, U2OS, KOLF2.1J, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a control matched to the primary antibody’s host and clonality: a subclass-matched isotype for a monoclonal antibody or host-matched nonimmune IgG for a polyclonal antibody (standard IHC practice). Use MRPL58 knockout material or a cognate-peptide block as a target-specific negative control; for chromogenic staining, check endogenous peroxidase and, if using the caption’s biotinylated secondary, endogenous biotin (standard IHC practice; selected-SKU caption: biotinylated secondary and DAB).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; the selected A31846-1 paraffin-section caption reports 1:25 dilution but no fixative (selected-SKU caption). The supplied evidence does not establish whether frozen sections or IF are easier than paraffin IHC; ICC-IF images support mitochondrial localization (HPA: mitochondria, supported). In adipose sections, lipid vacuoles can leave empty spaces that complicate assessment of cell-associated staining (standard IHC practice).
HPA tissue IHC evidence for MRPL58
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MRPL58 IHC Tips
Troubleshooting MRPL58 chromogenic IHC in paraffin sections, with one entry for secondary IF/ICC experiments.
How should I retrieve MRPL58 in paraffin sections with weak staining?
Start with citrate buffer at pH 6.0 for heat induced retrieval at 95–98 °C for 20 min (page retrieval specification). The selected A31846-1 paraffin colorectal carcinoma caption reports 1:25 antibody dilution and DAB detection, but states neither retrieval conditions nor fixative (selected A31846-1 caption). After cooling, compare glandular cell staining with a no primary control while keeping detection and counterstain constant (standard IHC practice; HPA: medium colon glandular staining). If staining remains weak, trial an alkaline buffer as a separately optimised fallback and assess tissue morphology and background alongside signal (standard IHC practice).
Could fixation explain inconsistent MRPL58 staining between sections?
Target specific MRPL58 sensitivity to fixation is unknown from the supplied evidence; the selected paraffin section caption does not state a fixative (selected A31846-1 caption). Record each specimen’s fixative, fixation duration, processing schedule, and section age before comparing staining intensity across batches (standard IHC practice). Test sections processed under different conditions together using the same citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval specification; standard IHC practice). Include a consistently processed reference section and a no primary control, then assess morphology and background before attributing an intensity difference to MRPL58 abundance (standard IHC practice).
Where should convincing MRPL58 chromogenic staining appear?
Expect predominantly cytoplasmic staining in tissue IHC (HPA: cytoplasmic expression in several tissues), consistent with mitochondrial localisation (UniProt Q14197: mitochondrion; HPA subcellular: supported mitochondrial location). In colon, compare glandular cells with nearby structures because HPA reports medium glandular staining there (HPA tissue IHC: colon glandular cells, Medium). A chromogenic cytoplasmic signal may not resolve individual mitochondria, so score the cellular compartment and tissue pattern rather than claiming organelle level resolution (standard IHC practice). Predominantly nuclear or crisp cell border staining warrants controls and closer review because those additional locations are uncertain in the subcellular record (HPA subcellular: nucleoplasm and plasma membrane uncertain).
How can I assess epitope related loss of MRPL58 staining?
Check whether the antibody recognises a region retained in the mature protein: MRPL58 is a 206 amino acid precursor with an annotated mature chain spanning residues 30–206 (UniProt Q14197: processing). The selected A31846-1 image describes a C terminal antibody, but does not map its exact epitope (selected A31846-1 caption). UniProt annotates 0 isoforms, no transmembrane segment, and an N5 methylglutamine at residue 90; these annotations alone do not establish epitope accessibility in sections (UniProt Q14197: isoforms, topology, modified residues). Compare retrieval conditions on matched sections and confirm any disputed pattern with an independently validated epitope when available (standard IHC practice).
How should IF/ICC help investigate an ambiguous IHC pattern?
Use secondary IF/ICC to examine whether MRPL58 overlaps a mitochondrial marker within the expected cells; HPA supports mitochondrial localisation and reports medium staining in colon glandular cells (HPA subcellular: mitochondria supported; HPA tissue IHC: colon glandular cells, Medium). Multiplex with a glandular epithelial cell marker to identify those cells, and include single stain controls to check channel bleed through (standard IF practice). Choose fluorophores after checking tissue autofluorescence, reserving a spectrally cleaner channel for weak MRPL58 signal (standard IF practice). Because MRPL58 has no annotated transmembrane segment and is mitochondrial, validate permeabilisation sufficient for antibody access to the intracellular epitope without assuming a specific mitochondrial membrane side (UniProt Q14197: topology and localisation; standard IF practice).
What causes diffuse or granular DAB background in MRPL58 IHC?
Run a no primary control and inspect whether DAB deposits follow tissue edges, damaged areas, or the expected cellular pattern (standard IHC practice). The selected A31846-1 example used a biotinylated secondary followed by DAB, so evaluate endogenous biotin and endogenous peroxidase as possible detection related background sources (selected A31846-1 caption; standard IHC practice). Apply an appropriate peroxidase block, check blocking and wash conditions, and titrate the antibody around the reported 1:25 example if nonspecific color persists (standard IHC practice; selected A31846-1 caption). Require reproducible cellular cytoplasmic staining before interpreting granular deposits as MRPL58, whose supported location is mitochondrial (HPA tissue IHC: cytoplasmic profile; HPA subcellular: mitochondria supported).
How should I quantify MRPL58 across IHC specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring; for colon sections, glandular cells are an evidence supported population to assess (HPA tissue IHC: colon glandular cells, Medium). Record the percentage of positive cells and staining intensity, or calculate an H score as the sum of intensity grade multiplied by its cell percentage (standard IHC practice). If counting positive cells per mm², normalise to viable tissue area and report the same compartment and threshold across specimens (standard IHC practice). Keep retrieval, antibody dilution, DAB development, and imaging settings consistent, and use a reference section to monitor batch drift (standard IHC practice).
How do I distinguish true MRPL58 signal from artefact?
Give greatest weight to reproducible cytoplasmic staining in intact cells, consistent with the tissue profile and supported mitochondrial localisation (HPA tissue IHC: cytoplasmic profile; HPA subcellular: mitochondria supported). Check whether the stained cell type matches the sampled tissue: HPA reports medium colon glandular staining but no detected signal in several other listed cell populations (HPA tissue IHC: positive and negative cell profiles). Discount isolated edge staining, necrotic deposits, and color retained in a no primary control; investigate endogenous peroxidase or biotin in DAB workflows (standard IHC practice; selected A31846-1 caption: biotinylated secondary and DAB). Treat discordance cautiously because HPA marks its tissue IHC reliability Approved while reporting low consistency with RNA expression (HPA tissue IHC: reliability description).
Boster reagents
Best MRPL58 / Large ribosomal subunit protein mL62 IHC Antibodies
A31846-1 has IHC images from human paraffin-embedded colorectal carcinoma and skin sections (catalog image captions). No IF/ICC images are provided (catalog payload).
Real IHC data
Anti-ICT1 Antibody (C-term)
Cat # A31846-1
A31846-1 is the sole SKU with a rendered card, showing human paraffin-embedded colorectal carcinoma IHC at 1:25 (card caption). Its catalog also shows human paraffin-embedded skin IHC at 1:25 (catalog image caption).
Which to pick: Choose polyclonal A31846-1 for human paraffin-section IHC because its own captions show staining in colorectal carcinoma and skin sections; the fixative is unreported (A31846-1 catalog and image captions). For cross-species IHC, A31846 lists human, mouse and rat reactivity and an IHC application, but provides no IHC image (A31846 catalog). Neither SKU lists IF/ICC or provides an IF image, so there is no catalog-supported IF/ICC pick (catalog payload).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q14197 (ICT1_HUMAN, Large ribosomal subunit protein mL62).
- Human Protein Atlas. MRPL58 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MRPL58 subcellular location (ICC-IF): Mainly localized to the mitochondria. In addition localized to the nucleoplasm and plasma membrane..
- Human Protein Atlas. MRPL58 antibody validation summary (2 antibodies).
- Identification of MAEA protein as a potential target for chemoresistance in osteosarcoma using bioinformatics and proteomic analysis. Frontiers in oncology 2025 — PMC12450695.
- PubMed PMID:8575443 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MRPL58 (Large ribosomal subunit protein mL62). Expected localisation Observed cytoplasm (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: A31846-1 at 1:25 (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Bone marrow. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded H. colorectal carcinoma tissue; fixative not specified (datasheet A31846-1); 2. Fixation: Image fixative and duration unreported (datasheet A31846-1); 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-MRPL58, 1:25 (datasheet A31846-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section 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 A31846-1); Caveat: Staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain spans 30–206 (UniProt); Regulation: Down-regulated during HT29-D4 differentiation (UniProt). Evidence: UniProt Q14197, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the mitochondria. In addition localized to the…, Datasheet A31846-1, PMC12450695. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MRPL58/immunohistochemistry-ihc-guide.