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MRPL12 / Large ribosomal subunit protein bL12m · IHC design guide
Design Immunohistochemistry for MRPL12
Plan chromogenic IHC for MRPL12 in paraffin sections using the catalog antibody (datasheet A10395-1: IHC). Use widespread granular cytoplasmic staining as a reference pattern, and interpret specificity cautiously because staining may reflect proteins from more than one gene (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MRPL12 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 | Granular cytoplasm (HPA tissue IHC); mitochondrial matrix (UniProt) | |
| Staining pattern | Widespread granular cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A10395-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (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 A10395-1) | |
| Caveat | Staining may reflect proteins from more than one gene (HPA tissue IHC) | |
| Regulation | Low tissue specificity; no stimulus-linked change stated (HPA tissue IHC; UniProt) | |
| Isoform / epitope | No isoforms; an aa 37–44 epitope may miss the short mature form (UniProt) |
Section 1
Recommended MRPL12 IHC & IF Protocols
Start with the catalog antibody’s IHC-P protocol (datasheet A10395-1), then compare the published LUAD tissue protocols (PMC11441236; PMC9917664).
| Sample | Paraffin-embedded human Colon cancer tissue; fixative not specified (datasheet A10395-1) |
| Fixation | Image fixative and duration unreported (datasheet A10395-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A10395-1); 20 min, 95–100 °C (standard) |
| 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-MRPL12, 1:50 recommended; image 1:200 (datasheet A10395-1) |
| 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 | MRPL12-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression with a granular pattern. No signal in the no-primary control. |
💡Decision noteTry heat-mediated Tris-EDTA retrieval at pH 9.0 first (datasheet A10395-1); neither published excerpt specifies a retrieval method (PMC11441236; PMC9917664).
Section 2
What Is the Expected MRPL12 Staining Pattern?
MRPL12 is a mitochondrial matrix protein without a transmembrane segment (UniProt P52815). In paraffin tissue IHC, expect granular cytoplasmic staining across many cell types, with strong staining reported in selected glandular, epithelial, hematopoietic and neuronal cells (HPA: tissue IHC). The tissue profile is Supported, with medium consistency between staining and RNA data; HPA cautions that the antibodies target proteins from more than one gene (HPA: tissue IHC).
| Fine granular cytoplasmic staining in adrenal glandular cells or bronchial respiratory epithelial cells. | This fits MRPL12's mitochondrial matrix location and HPA's ubiquitous granular cytoplasmic profile; both listed cell types stain High (UniProt P52815; HPA: tissue IHC). Judge the pattern within the identified cells, since a strong chromogen deposit alone cannot establish which protein the antibody detected (HPA: multi-gene antibody caution). |
| Predominantly nuclear, cell-surface or extracellular staining, without a granular cytoplasmic component. | Those compartments conflict with the mitochondrial matrix assignment and the reported tissue pattern (UniProt P52815; HPA: tissue IHC). Treat the result as suspect and compare it with a known-positive section and negative detection control; compartment alone does not identify the source of an artefact (standard IHC practice). |
| Strong staining in an unexpected cell population while the expected granular pattern is absent. | Check cell identity and the HPA tissue profile before calling it abnormal: expression is widespread, and the supplied tissue record lists no negative cell population (HPA: tissue IHC). If staining follows the detection reagents or lacks the expected pattern, consider cross-reactivity or endogenous detection activity (standard IHC practice; HPA: multi-gene antibody caution). |
| Uniform haze across tissue and empty spaces, obscuring individual cytoplasmic granules. | A diffuse deposit is difficult to reconcile with HPA's granular cytoplasmic profile (HPA: tissue IHC). Compare a section without primary antibody, then review blocking, washes and chromogen development as general background controls; this observation does not establish an MRPL12-specific fixation effect (standard IHC practice). |
| No detectable staining in adrenal glandular cells or bronchial respiratory epithelial cells. | Both are reported High in HPA tissue IHC, so an absent signal warrants a technical check before a biological interpretation (HPA: tissue IHC). Verify that the positive-control tissue and detection reagents worked, then review the antibody's IHC-P instructions; the supplied sources do not establish target-specific retrieval or fixation sensitivity (standard IHC practice). |
💡Expected MRPL12 appearanceCall a positive result when identified cells show granular cytoplasmic staining consistent with mitochondria, especially in HPA High populations; isolated nuclear or diffuse extracellular colour is suspect (UniProt P52815; HPA: tissue IHC).
| Mitochondrial location and soluble topology | MRPL12 is assigned to the matrix and has no transmembrane segment (UniProt P52815). In IHC, assess intracellular granular cytoplasmic staining; membrane-rim staining has no support from the supplied localisation records (UniProt P52815; HPA: tissue IHC). |
| Cell population and tissue context | HPA reports Low tissue RNA specificity and a ubiquitous granular cytoplasmic protein profile, yet selected populations stain High and skeletal myocytes stain Low (HPA: tissue IHC). A weak myocyte result is therefore a poor basis for declaring a failed stain (HPA: tissue IHC). |
| Antibody attribution | Both listed antibodies have Supported IHC status, while HPA reports medium staining–RNA consistency and warns that the antibodies target proteins from more than one gene (HPA: antibody validation; HPA: tissue IHC). Pattern agreement supports interpretation but cannot, by itself, prove MRPL12-specific binding (HPA: multi-gene antibody caution). |
| Protein maturation | UniProt lists a 198-residue precursor and mature chains spanning residues 37–198 or 45–198 (UniProt P52815). Interpret staining as cellular localisation; without a supplied epitope map, these processing annotations cannot predict which form the IHC antibody detects (UniProt P52815). |
| IF/ICC comparison | For the separate IF/ICC guide: HPA reports supported mitochondrial localisation in ICC-IF and images from A-431, U-251MG and U2OS; both listed antibodies have Supported ICC status (HPA: subcellular; HPA: antibody validation). This supports a localisation cross-check, subject to HPA's multi-gene antibody caution (HPA: subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive-control section is blank. | A run-level antibody, detection or development failure is possible; adrenal glandular and bronchial respiratory epithelial cells are reported High (standard IHC practice; HPA: tissue IHC). | Check the control slide, reagent sequence and IHC-P instructions, then repeat the run with a documented positive control (standard IHC practice). Do not infer an MRPL12-specific fixation or retrieval effect from the supplied records. |
| The test section is weak, but the positive control stains. | Cell composition may explain the difference: HPA lists skeletal myocytes as Low, versus High staining in several glandular, epithelial, hematopoietic and neuronal populations (HPA: tissue IHC). | Confirm which cells were scored and compare like cell populations across sections (standard IHC practice). Report weak staining in its tissue context rather than treating every weak section as technical failure (HPA: tissue IHC). |
| Colour appears mainly over nuclei or along cell borders. | That distribution disagrees with the mitochondrial matrix assignment and HPA's granular cytoplasmic profile (UniProt P52815; HPA: tissue IHC). | Recheck focus, counterstain and cell boundaries; compare a known-positive section and a no-primary control before scoring the signal (standard IHC practice). |
| Granules are lost in widespread brown haze. | Background from detection reagents, incomplete blocking or excessive development can obscure cellular detail (standard IHC practice). | Inspect the no-primary control and review blocking, washes and development against the established IHC workflow (standard IHC practice). Score only a resolved cellular pattern that matches the reported granular cytoplasm (HPA: tissue IHC). |
| An unexpected cell population stains strongly. | MRPL12's tissue profile is broad, but antibody cross-reactivity or endogenous detection activity can also produce apparent signal (HPA: tissue IHC; standard IHC practice). HPA specifically cautions that its antibodies target proteins from more than one gene (HPA: tissue IHC). | Identify the stained cells, check whether signal is granular and cytoplasmic, and compare the no-primary control and HPA tissue observations (standard IHC practice; HPA: tissue IHC). Avoid assigning unexpected staining to MRPL12 from intensity alone (HPA: multi-gene antibody caution). |
| IHC granules and an IF/ICC image appear different. | The records describe granular cytoplasm in tissue IHC and supported mitochondrial localisation in ICC-IF; the methods display cellular detail differently (HPA: tissue IHC; HPA: subcellular; standard IHC practice). | Compare compartment and cell identity rather than requiring identical visual texture (standard IHC practice). Use the separate IF/ICC guide for IF decisions; HPA's multi-gene antibody caution applies to its localisation images (HPA: subcellular). |
Experiment evidence
IHC & IF images showing MRPL12 staining
A10395-1 · IHCImmunohistochemical analysis of paraffin-embedded human Colon cancer. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 w
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000262814-MRPL12/tissue/cerebral+cortex#imid_6016424
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000262814-MRPL12/tissue/colon#imid_6015134
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000262814-MRPL12/tissue/liver#imid_6015119
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000262814-MRPL12/tissue/kidney#imid_6015128
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA022853; green = MRPL12, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA022853; green = MRPL12, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MRPL12 IHC & IF
🧪Run bone marrow first and assess MRPL12 staining in hematopoietic cells (HPA: High in hematopoietic cells). HPA detects MRPL12 in all 45 scored tissues, so use no-primary and isotype controls for the negative; other cells on the positive slide should be judged against control background, not assumed negative (HPA: no negative tissue listed; standard IHC control practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: MRPL12 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MRPL12 in A-431, U-251MG, U2OS, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide, an isotype control matched to the primary antibody’s host species and immunoglobulin class, and a biological negative such as knockout material or peptide blocking if the immunizing peptide is available (standard IHC control practice). Quench endogenous peroxidase in bone marrow and check for endogenous biotin if using avidin–biotin detection, since either can produce chromogenic background (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported; the selected A10395-1 paraffin-section caption does not state a fixative (selected-SKU tissue-IHC caption: fixative unreported). That caption uses Tris-EDTA at pH 9.0 for retrieval and a 1:200 primary dilution, but it does not establish that retrieval is required (selected-SKU tissue-IHC caption). The supplied evidence does not establish whether frozen sections or IF are easier; for bone marrow IHC, assess endogenous peroxidase background with the controls (HPA: High in hematopoietic cells; standard IHC practice).
HPA tissue IHC evidence for MRPL12
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: MRPL12 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced MRPL12 IHC Tips
Troubleshoot MRPL12 staining in paraffin sections by checking retrieval, granular cytoplasmic localisation, controls, and scoring consistency (UniProt P52815; HPA tissue IHC).
Which retrieval conditions should I try first for weak MRPL12 staining?
Use heat-mediated Tris-EDTA pH 9.0 antigen retrieval for this paraffin-section IHC assay (datasheet A10395-1). The selected colon cancer image used that buffer, followed by primary antibody at 1:200 overnight at 4°C (caption A10395-1). If staining remains weak, compare a citrate pH 6.0 retrieval condition on matched sections as a fallback, keeping detection and imaging settings consistent (standard IHC practice). Include a no-primary control and inspect tissue morphology, because stronger chromogen after harsher retrieval alone does not establish specific MRPL12 staining (standard IHC practice).
How should I troubleshoot possible fixation effects on MRPL12 IHC?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative, so its staining cannot establish which fixation conditions preserve MRPL12 (caption A10395-1). Record the actual fixative, fixation duration, processing schedule, and section age for each specimen before comparing staining (standard IHC practice). Apply the documented Tris-EDTA pH 9.0 retrieval and 1:200 primary dilution consistently across a small matched comparison (caption A10395-1). If signal differs, check morphology and no-primary controls, then report the difference as condition dependent without assigning a target-specific fixation mechanism (standard IHC practice).
What staining pattern is plausible for MRPL12 in paraffin sections?
Look for granular cytoplasmic staining, consistent with the reported tissue pattern and mitochondrial localisation of MRPL12 (HPA tissue IHC; UniProt P52815 subcellular location). MRPL12 is assigned to the mitochondrial matrix and has no transmembrane segment, so a crisp plasma-membrane outline is unexpected (UniProt P52815 topology). The atlas reports high staining in adrenal glandular cells and low staining in skeletal-muscle myocytes; use these as comparative observations, not absolute positive and negative controls (HPA tissue IHC). Examine staining at the same magnification and chromogen exposure across sections, with a no-primary control to assess nonspecific granular deposits (standard IHC practice).
Could MRPL12 processing or modifications explain inconsistent staining?
MRPL12 has no annotated isoforms, but its mature protein is reported in long 37–198 and short 45–198 forms (UniProt P52815 processing). An antibody epitope within residues 37–44 could therefore distinguish those forms; epitope information is needed before attributing a staining difference to processing (UniProt P52815 processing; standard IHC interpretation). Several lysine acetylation sites are annotated, including positions 125, 138, 142, and 144, without supplied evidence that they alter this antibody’s IHC binding (UniProt P52815 modified residues). Compare matched retrieval conditions and an independently validated antibody before proposing epitope masking or a biological change (standard IHC practice).
How can I assess MRPL12 by multiplex IF alongside tissue IHC?
Treat IF as a separately validated assay: the selected catalog evidence documents paraffin-section chromogenic IHC, with pH 9.0 retrieval, but supplies no IF procedure for that antibody (caption A10395-1). For a tissue comparison, multiplex with a marker identifying the expected cell population, such as adrenal glandular cells, and assess whether MRPL12 appears in their cytoplasm (HPA tissue IHC; standard IF practice). Prefer a far-red fluorophore when tissue autofluorescence obscures shorter wavelengths, and include single-channel and no-primary controls (standard IF practice). Because MRPL12 resides in the mitochondrial matrix, optimise permeabilisation to allow antibody access across mitochondrial membranes while preserving morphology; verify the pattern independently (UniProt P52815 subcellular location; standard IF practice).
What should I check when MRPL12 staining looks diffuse or dirty?
First compare the section with a no-primary control and inspect whether colour follows tissue edges, folds, pigment, or damaged areas (standard IHC practice). Quench endogenous peroxidase for enzyme-based detection and check whether the secondary reagent alone produces deposits; these are general workflow checks, not MRPL12-specific findings (standard IHC practice). The selected image used primary and secondary dilutions of 1:200, with secondary incubation for 45 minutes at room temperature; use those as documented starting conditions for the same assay (caption A10395-1). Reduce nonspecific staining by checking blocking, washing, antibody concentration, and chromogen development while retaining expected granular cytoplasmic signal (HPA tissue IHC; standard IHC practice).
How should I score MRPL12 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score granular cytoplasmic staining within that population, rather than treating all coloured area as MRPL12 positive (HPA tissue IHC; standard IHC practice). Record the percentage of positive cells and intensity, then calculate an H-score on a consistent 0–300 scale when intensity categories are reproducible (standard IHC practice). Alternatively, report positive-cell density per mm² of viable tissue, normalised to the measured viable area and accompanied by the number of evaluable cells (standard IHC practice). Keep retrieval, detection, counterstain, image acquisition, and scoring thresholds consistent; exclude necrosis and folds using predefined rules (standard IHC practice).
When should I doubt an apparent MRPL12-positive IHC result?
A convincing result shows granular cytoplasmic staining in intact cells, consistent with the reported tissue profile and mitochondrial matrix assignment (HPA tissue IHC; UniProt P52815 subcellular location). Predominantly nuclear or plasma-membrane staining, signal confined to tissue edges or necrosis, and colour in no-primary controls warrant investigation before interpretation (UniProt P52815 topology; standard IHC practice). Check endogenous peroxidase and pigment when deposits appear independent of cellular granules, and compare staining in the selected cell population across matched sections (standard IHC practice). The atlas rates tissue staining as supported but cautions that its antibodies target proteins from multiple genes, so atlas concordance alone cannot prove specificity for this assay (HPA tissue IHC).
Boster reagents
Best MRPL12 / Large ribosomal subunit protein bL12m IHC Antibodies
MRPL12 IHC is listed for human and mouse samples (catalog applications/reactivity), with a paraffin-section human colon cancer image for A10395-1 (image caption). A10395-3 also lists IF/ICC (catalog applications).
Real IHC data
Anti-MRP-L12 Antibody
Cat # A10395-1
A10395-1 lists human and mouse IHC and shows staining in paraffin-embedded human colon cancer (catalog applications/reactivity; A10395-1 image caption). A10395-3 lists human, mouse, and rat reactivity for an antibody with IHC and IF/ICC applications, but has no IHC or IF image in the payload (A10395-3 catalog applications/reactivity; image alts).
Which to pick: For paraffin-section tissue IHC, choose A10395-1: its own image caption documents human colon cancer sections, 1:200 primary dilution, and Tris-EDTA retrieval at pH 9.0; the fixative is unreported (A10395-1 image caption). For IF/ICC, choose A10395-3, which lists both applications and a 1:50 IF dilution, though no IF image is supplied (A10395-3 catalog applications/dilution; image alts). For rat samples, A10395-3 is the listed option because its reactivity includes rat; it is polyclonal (A10395-3 catalog reactivity; dilution_raw).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P52815 (RM12_HUMAN, Large ribosomal subunit protein bL12m).
- Human Protein Atlas. MRPL12 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. MRPL12 subcellular location (ICC-IF): Localized to the mitochondria. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. MRPL12 antibody validation summary (2 antibodies).
- MRPL12 K163 acetylation inhibits ccRCC via driving mitochondrial metabolic reprogramming. Cell death & disease 2025 — PMC12381009.
- UBASH3B-mediated MRPL12 Y60 dephosphorylation inhibits LUAD development by driving mitochondrial metabolism reprogramming. Journal of experimental & clinical cancer research : CR 2024 — PMC11441236.
- MRPL12 Acts as A Novel Prognostic Biomarker Involved in Immune Cell Infiltration and Tumor Progression of Lung Adenocarcinoma. International journal of molecular sciences 2023 — PMC9917664.
- Mitochondrial ribosomal protein L12 mediates metabolic reorganization in clear cell renal cell carcinoma by regulating mitochondrial biosynthesis. Cell communication and signaling : CCS 2025 — PMC12523095.
- PubMed PMID:8626705 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:26586915 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MRPL12 (Large ribosomal subunit protein bL12m). Expected localisation Granular cytoplasm (HPA tissue IHC); mitochondrial matrix (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A10395-1). Validated IHC antibody: A10395-1 at 1:50 recommended; image 1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human Colon cancer tissue; fixative not specified (datasheet A10395-1); 2. Fixation: Image fixative and duration unreported (datasheet A10395-1); 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 retrieval: Tris-EDTA pH 9.0 (datasheet A10395-1); 20 min, 95–100 °C (standard); 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-MRPL12, 1:50 recommended; image 1:200 (datasheet A10395-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (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 A10395-1); Caveat: Staining may reflect proteins from more than one gene (HPA tissue IHC); Isoform / epitope: No isoforms; an aa 37–44 epitope may miss the short mature form (UniProt); Regulation: Low tissue specificity; no stimulus-linked change stated (HPA tissue IHC; UniProt). Evidence: UniProt P52815, HPA tissue IHC · Supported, HPA ICC-IF · mitochondria. Caution: Based on antibodies targeting proteins from…, Datasheet A10395-1, PMC11441236, PMC9917664. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MRPL12/immunohistochemistry-ihc-guide.