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LONP1 / Lon protease homolog, mitochondrial · IHC design guide
Design Immunohistochemistry for LONP1
Plan paraffin-section LONP1 IHC around granular cytoplasmic staining in glandular cells (HPA tissue IHC) and its mitochondrial matrix location (UniProt). Use adrenal glandular cells as a high-staining reference and thyroid glandular cells as a not-detected reference (HPA tissue IHC); the IHC-validated antibody is listed at 0.5–1 μg/mL (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LONP1 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 | Granular cytoplasmic staining in glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A03808-2) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Thyroid gland |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Thyroid glandular cells are not detected, despite positive glands elsewhere (HPA tissue IHC) | |
| Regulation | Adrenal gland RNA is tissue enhanced (HPA tissue RNA) | |
| Isoform / epitope | 3 isoforms; mature chain 68–959; epitope impact unknown (UniProt) |
Section 1
Recommended LONP1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses citrate pH 6 retrieval (datasheet A03808-2). One published LONP1 tumor-section protocol follows (PMC11840119).
| Sample | Paraffin-embedded human mammary cancer tissues; fixative not specified (datasheet A03808-2) |
| Fixation | Image fixative and duration unreported (datasheet A03808-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A03808-2) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03808-2) |
| Primary antibody | Rabbit anti-LONP1, 0.5-1μg/ml (datasheet A03808-2) |
| Primary incubation | Overnight at 4 °C (datasheet A03808-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03808-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LONP1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Granular cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteTry heat-mediated citrate pH 6 retrieval first; both the catalog datasheet and the published protocol specify it (datasheet A03808-2; PMC11840119).
Section 2
What Is the Expected LONP1 Staining Pattern?
LONP1 is a mitochondrial matrix protein with no transmembrane segment (UniProt P36776). In paraffin-section IHC, expect granular cytoplasmic staining, especially in adrenal, duodenal, appendiceal and colonic glandular cells and bronchial respiratory epithelium (HPA: granular cytoplasmic profile; High in listed cells). HPA rates its tissue staining Supported, citing consistency with RNA expression (HPA: Supported).
| Granular cytoplasmic staining in adrenal or duodenal glandular cells. | This fits the reported pattern and provides a useful positive reference (HPA: High in these cells; granular cytoplasmic profile). Score the relevant cells and their cytoplasmic distribution; staining of a tissue section alone does not establish that every cell is positive (general IHC practice). |
| Predominantly nuclear or cell-surface staining, without a convincing granular cytoplasmic component. | Question the signal's specificity: LONP1 resides in the mitochondrial matrix and has no transmembrane segment (UniProt P36776). HPA reports uncertain nucleoplasmic and cytosolic signals in ICC-IF; those observations do not establish a predominantly nuclear or surface IHC pattern (HPA: subcellular ICC-IF). |
| Strong signal in a cell population where the HPA tissue profile reports little or no staining. | Compare the precise cell type before calling it positive. Thyroid glandular cells are Not detected, while heart cardiomyocytes and liver cholangiocytes are Low in the supplied IHC profile (HPA: tissue IHC). Unexpected strong staining warrants checks for cross-reactivity or endogenous detection activity (general IHC practice). |
| Diffuse chromogen across tissue, including spaces or structures without identifiable cells. | A widespread deposit is difficult to reconcile with the reported granular cytoplasmic pattern (HPA: tissue IHC). Check background in the relevant controls and reassess blocking, washing and detection reagents before interpreting cell-specific LONP1 expression (general IHC practice). |
| No detectable signal in adrenal or duodenal glandular cells. | These are reported High in the HPA IHC profile, so a blank result calls for a technical review before a biological negative call (HPA: High in these cells). Inspect section quality, antigen retrieval, antibody use and chromogen development as general IHC checks; target-specific fixation sensitivity is unreported here. |
💡Expected LONP1 appearanceCall a positive result when glandular or respiratory epithelial cells show distinct granular cytoplasmic staining, potentially High in the listed positive tissues (HPA: tissue IHC); broad nuclear, surface or tissue-wide diffuse staining should prompt specificity checks (UniProt P36776; HPA: granular cytoplasmic profile; general IHC practice).
| Cell and tissue context | HPA reports High staining in adrenal, appendix, colon and duodenal glandular cells and bronchial respiratory epithelium; thyroid glandular cells are Not detected (HPA: tissue IHC). These are cell-specific observations, so compare like cells when choosing a reference section (general IHC practice). |
| Subcellular interpretation | Matrix localisation and the absence of a transmembrane segment support a cytoplasmic, mitochondrial interpretation (UniProt P36776). HPA describes the tissue-IHC profile as granular cytoplasmic; routine chromogenic IHC does not by itself prove mitochondrial colocalisation (HPA: tissue IHC; general IHC practice). |
| Antibody validation | HPA002192 carries an IHC Supported rating; HPA002034 has no supplied IHC rating (HPA: antibody validation). Do not transfer the antibodies' ICC Enhanced ratings to paraffin IHC (HPA: antibody validation). Interpret an unexpected pattern alongside appropriate assay controls (general IHC practice). |
| Processing and isoforms | UniProt lists 3 isoforms and a mature chain spanning residues 68–959 (UniProt P36776). Epitope-dependent differences are possible in principle, but no epitope position or isoform-specific IHC result is supplied; do not assign a particular staining pattern to either feature. |
| IF/ICC Q&A | What location should IF/ICC show? Mainly mitochondria; HPA also lists nucleoplasm and cytosol as uncertain additional locations (HPA: subcellular ICC-IF). This localisation answer is not an IF/ICC protocol or a substitute for interpreting the paraffin-section IHC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive reference tissue is blank. | A technical step may have failed; adrenal and duodenal glandular cells are reported High (HPA: tissue IHC). The supplied sources do not identify a LONP1-specific fixation effect. | Review section integrity, retrieval, antibody application and detection development with assay controls (general IHC practice). Avoid interpreting the blank section as biological absence until those checks pass. |
| Signal is mostly nuclear. | This conflicts with mitochondrial matrix localisation and the granular cytoplasmic tissue-IHC profile (UniProt P36776; HPA: tissue IHC). HPA's nucleoplasmic ICC-IF location is uncertain (HPA: subcellular ICC-IF). | Recheck morphology and background controls; compare with a known High cell population in the same staining workflow (HPA: tissue IHC; general IHC practice). |
| Thyroid glandular cells stain strongly. | HPA reports this cell population as Not detected, making strong signal unexpected for the supplied tissue-IHC reference (HPA: thyroid glandular cells). Cross-reactivity or endogenous detection activity is possible (general IHC practice). | Verify cell identity and inspect reagent-control staining; reassess blocking and detection if controls also develop chromogen (general IHC practice). |
| Weak staining in cardiomyocytes is scored as a failed assay. | HPA reports Low staining in cardiomyocytes, although UniProt lists heart among tissues expressing LONP1 (HPA: heart muscle; UniProt P36776 tissue specificity). These statements address different levels of evidence. | Use a reported High cell population to judge technical performance, then record cardiomyocyte intensity separately (HPA: tissue IHC; general IHC practice). |
| Granules are obscured by diffuse background. | Excess background can mask the reported granular cytoplasmic pattern (HPA: tissue IHC; general IHC practice). The supplied sources do not establish a LONP1-specific retrieval or fixation cause. | Inspect control sections and review blocking, washing and chromogen development as general IHC workflow checks before scoring localisation (general IHC practice). |
| Two antibodies appear to give different IHC patterns. | The supplied HPA ratings differ: HPA002192 is IHC Supported, while HPA002034 has no supplied IHC rating; both are ICC Enhanced (HPA: antibody validation). Their ICC status cannot resolve an IHC discrepancy. | Compare cell-level localisation against the HPA granular cytoplasmic profile and assess each IHC assay with its own controls (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing LONP1 staining
A03808-2 · IHCIHC analysis of LONP1 using anti-LONP1 antibody (A03808-2). LONP1 was detected in paraffin-embedded section of human mammary cancer tissues.
A03808-2 · IHCIHC analysis of LONP1 using anti-LONP1 antibody (A03808-2). LONP1 was detected in paraffin-embedded section of human rectal cancer tissues.
A03808-2 · IFIF analysis of LONP1 using anti-LONP1 antibody (A03808-2). LONP1 was detected in immunocytochemical section of A431 cells. Enzyme antigen re
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000196365-LONP1/tissue/cerebral+cortex#imid_911072
HPA · DuodenumHuman Protein Atlas IHC, Duodenum. Source: https://www.proteinatlas.org/ENSG00000196365-LONP1/tissue/duodenum#imid_910997
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000196365-LONP1/tissue/colon#imid_910998
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000196365-LONP1/tissue/liver#imid_910991
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA002034; green = LONP1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA002034; green = LONP1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LONP1 IHC & IF
🧪Run duodenum first: its glandular cells should stain for LONP1 (HPA: High in duodenal glandular cells). Run thyroid gland as the negative tissue (HPA: Not detected in thyroid glandular cells); on the duodenal slide, unstained non-glandular cells can help assess background, but their LONP1-negative status is unverified (HPA: High is specified for glandular cells).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Thyroid gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LONP1 in A-431, U-251MG, U2OS, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a rabbit IgG control matched to the primary antibody format, and LONP1-knockout material or a validated immunogen-peptide block where available (selected tissue-IHC caption: rabbit primary; standard IHC practice). For chromogenic detection, block endogenous peroxidase and check for endogenous biotin background when using the caption’s streptavidin-biotin system (selected tissue-IHC caption: SABC with DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU tissue-IHC caption does not report a fixative (A03808-2 tissue-IHC caption). The caption reports heat-mediated citrate retrieval at pH 6 for 20 minutes, which is a documented condition rather than proof that retrieval is required (A03808-2 tissue-IHC caption). For IF/ICC, mitochondrial enrichment is the supported expected pattern, but the supplied evidence does not establish whether IF or frozen sections are easier; in duodenum, check chromogenic signal for endogenous biotin background (HPA: mitochondria supported; UniProt P36776: mitochondrial matrix; selected tissue-IHC caption: SABC detection; standard IHC practice).
HPA tissue IHC evidence for LONP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — High 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Thyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LONP1 IHC Tips
Troubleshoot LONP1 staining in paraffin sections by checking retrieval, compartment, cell type and controls before comparing chromogenic signal across specimens.
Which retrieval conditions should I start with for LONP1 in paraffin sections?
Start with heat-mediated citrate buffer at pH 6 for 20 minutes (datasheet A03808-2). The selected paraffin-section example used these conditions, followed by 10% goat-serum block and 1 μg/ml primary antibody overnight at 4°C (datasheet A03808-2). If staining is weak, check deparaffinisation, heating consistency and tissue preservation before changing retrieval conditions (standard IHC practice). Compare any adjusted retrieval condition on adjacent sections with the same antibody concentration and detection workflow, because stronger DAB signal can also reflect increased background (standard IHC practice). Retain the condition that preserves morphology and granular cytoplasmic staining (HPA: tissue IHC profile).
How should I troubleshoot fixation when LONP1 staining is weak or uneven?
The selected paraffin-section caption does not state a fixative, so LONP1-specific sensitivity to fixation is unknown (datasheet A03808-2). Record the fixative and fixation duration for each specimen, then compare sections processed and stained together (standard IHC practice). Check whether weak areas coincide with poor morphology or uneven staining across the section before changing antibody concentration (standard IHC practice). Keep the documented citrate pH 6, 20-minute retrieval constant while assessing processing differences (datasheet A03808-2). Use the same positive-control tissue in each run so a global staining failure is easier to distinguish from a specimen-specific problem (standard IHC practice).
What LONP1 staining pattern should I expect in chromogenic tissue sections?
Expect granular cytoplasmic staining in tissue sections (HPA: tissue IHC profile), consistent with LONP1 in the mitochondrial matrix (UniProt P36776: subcellular location). Its lack of a transmembrane segment does not support a crisp plasma-membrane pattern (UniProt P36776: topology). Compare the observed pattern with glandular cells in adrenal gland or duodenum, where staining is reported as high (HPA: tissue IHC). Interpret faint diffuse cytoplasmic colour cautiously when granules are absent, and inspect the matching negative control (standard IHC practice). Nucleoplasmic and cytosolic locations have uncertain support in cell imaging, so neither alone establishes specific chromogenic tissue staining (HPA: subcellular location).
Can this stain distinguish LONP1 isoforms or modified epitopes?
LONP1 has 3 listed isoforms, but the supplied tissue-IHC caption does not identify an antibody epitope or establish isoform selectivity (UniProt P36776: isoforms; datasheet A03808-2). Its annotated chain spans residues 68–959, with a Lon N-terminal domain at 124–370 and proteolytic domain at 759–949 (UniProt P36776: processing and domains). Phosphotyrosines at residues 186 and 394 are annotated, but their effect on this stain is untested here (UniProt P36776: modified residues; datasheet A03808-2). Request the immunogen or epitope specification before assigning a staining difference to an isoform or modification (standard IHC practice). Compare specimens under identical retrieval and detection conditions first (standard IHC practice).
How can I use IF to investigate an ambiguous LONP1 IHC pattern?
Use IF as a separate validation experiment: the supplied product example documents paraffin-section chromogenic IHC, with no IF conditions specified (datasheet A03808-2). Multiplex LONP1 with a marker for the expected cell type, such as glandular cells in adrenal gland or duodenum, and assess staining within those cells (HPA: tissue IHC; standard IF practice). Add a mitochondrial marker to evaluate the expected compartment, while treating any apparent nuclear signal cautiously (UniProt P36776: mitochondrial matrix; HPA: nucleoplasm uncertain). Choose fluorophores and controls that separate specific signal from tissue autofluorescence (standard IF practice). Because the target is in the matrix, optimise permeabilisation for access across mitochondrial membranes without losing morphology (UniProt P36776: subcellular location; standard IF practice).
How do I reduce diffuse or brown background in LONP1 IHC?
First compare the stained section with a primary-antibody omission control and inspect whether colour follows tissue edges, damaged regions or the expected cellular granules (standard IHC practice; HPA: tissue IHC profile). The selected example used 10% goat-serum block and 1 μg/ml primary antibody overnight at 4°C (datasheet A03808-2). For DAB detection, include an endogenous-peroxidase block and check secondary-antibody background as general chromogenic IHC controls (standard IHC practice). If background remains high, titrate the primary antibody and adjust blocking or washes while keeping retrieval and exposure consistent across comparison sections (standard IHC practice). Judge improvement by retained granular cytoplasmic signal, not merely paler tissue (HPA: tissue IHC profile).
How should I score LONP1 staining across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and regions before scoring; glandular cells can show high LONP1 staining in adrenal gland and duodenum (HPA: tissue IHC). For chromogenic sections, record the percentage of positive cells and staining intensity, then calculate an H-score with a maximum of 300 if the study uses the conventional 0–3 intensity scale (standard IHC practice). Alternatively, measure positive-cell density per mm² of viable tissue (standard IHC practice). Normalise comparisons to the same cell population or viable tissue area, and keep retrieval, detection and imaging settings consistent (standard IHC practice). Score granular cytoplasmic signal separately from diffuse background (HPA: tissue IHC profile; standard IHC practice).
When is an apparent LONP1-positive area more likely to be artefact?
A convincing positive pattern is granular cytoplasmic staining in intact cells, matching the tissue profile and mitochondrial matrix location (HPA: tissue IHC profile; UniProt P36776: subcellular location). Compare cell identity: adrenal and duodenal glandular cells are reported high, whereas thyroid glandular cells are reported not detected (HPA: tissue IHC). Treat isolated membrane-like or nuclear staining cautiously; LONP1 has no transmembrane segment, and its nucleoplasmic location has uncertain support (UniProt P36776: topology; HPA: subcellular location). Inspect section edges and necrotic areas for uneven deposits, and compare a primary-antibody omission control (standard IHC practice). For DAB staining, check endogenous-peroxidase blocking before interpreting residual brown colour as target signal (standard IHC practice).
Boster reagents
Best LONP1 / Lon protease homolog, mitochondrial IHC Antibodies
The catalog antibody has IHC data from human paraffin sections and IF/ICC data from A431 cells (image captions); listed reactivity includes human, mouse and rat (catalog: reactivity).
Real IHC data
Anti-LONP1/Lon Antibody ®
Cat # A03808-2
A03808-2 will render with IHC images from human mammary, rectal and lung cancer paraffin sections (A03808-2: IHC image captions). Its IF/ICC image shows staining in A431 cells (A03808-2: IF image caption).
Which to pick: Choose A03808-2 for paraffin-section IHC: its images show human tissue staining after citrate retrieval, while the fixative is unreported (A03808-2: IHC image captions). The same rabbit-host antibody is listed for IF/ICC and has an A431 cell image; its clonality is unreported (A03808-2: catalog applications, host and clone; IF image caption). For mouse or rat samples, A03808-2 lists reactivity with both species, but the supplied IHC images show human tissue only (A03808-2: catalog reactivity; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P36776 (LONM_HUMAN, Lon protease homolog, mitochondrial).
- Human Protein Atlas. LONP1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. LONP1 subcellular location (ICC-IF): Mainly localized to the mitochondria. In addition localized to the nucleoplasm and cytosol..
- Human Protein Atlas. LONP1 antibody validation summary (2 antibodies).
- Complement protein C1q induces endothelial apoptosis through C1qBP regulation of mitochondrial function in lung endothelial cells in systemic sclerosis. Molecular medicine (Cambridge, Mass.) 2026 — PMC13069790.
- LONP1 alleviates ageing-related renal fibrosis by maintaining mitochondrial homeostasis. Journal of cellular and molecular medicine 2024 — PMC11390342.
- Akt phosphorylation of mitochondrial Lonp1 protease enables oxidative metabolism and advanced tumor traits. Oncogene 2019 — PMC6814529.
- Lon protease 1-mediated metabolic reprogramming promotes the progression of prostate cancer. Cell death & disease 2025 — PMC11840119.
- PubMed PMID:8248235 — UniProt-cited evidence.
- PubMed PMID:20843780 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LONP1 (Lon protease homolog, mitochondrial). Expected localisation Granular cytoplasm (HPA tissue IHC); mitochondrial matrix (UniProt). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A03808-2). Validated IHC antibody: A03808-2 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Thyroid gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human mammary cancer tissues; fixative not specified (datasheet A03808-2); 2. Fixation: Image fixative and duration unreported (datasheet A03808-2); 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: Citrate pH 6 , 20 min (datasheet A03808-2); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03808-2); 8. Primary antibody: Rabbit anti-LONP1, 0.5-1μg/ml (datasheet A03808-2); 9. Primary incubation: Overnight at 4 °C (datasheet A03808-2). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Thyroid glandular cells are not detected, despite positive glands elsewhere (HPA tissue IHC); Isoform / epitope: 3 isoforms; mature chain 68–959; epitope impact unknown (UniProt); Regulation: Adrenal gland RNA is tissue enhanced (HPA tissue RNA). Evidence: UniProt P36776, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the mitochondria. In addition localized to the…, Datasheet A03808-2, PMC11840119. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LONP1/immunohistochemistry-ihc-guide.