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MLIP / Muscular LMNA-interacting protein · IHC design guide
Design Immunohistochemistry for MLIP
Plan MLIP staining in paraffin sections using heart and skeletal muscle as positive tissues (HPA tissue IHC). The guide covers fixation consistency, antibody setup and interpretation of predominantly cytoplasmic muscle staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MLIP 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 | Muscle cytoplasm (HPA tissue IHC); nuclear sites (UniProt) | |
| Staining pattern | High cytoplasmic staining in cardiomyocytes and skeletal myocytes (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Heart muscle+2 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 | IHC/RNA concordance is medium; verify with controls (HPA tissue IHC) | |
| Regulation | Heart and skeletal muscle enriched (UniProt) | |
| Isoform / epitope | 4 isoforms; confirm antibody epitope coverage (UniProt) |
Section 1
Recommended MLIP IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by 1 published MLIP staining protocol (PMC3103349).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A12261); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-MLIP, 5 μg/mL (datasheet A12261) |
| 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 | MLIP-positive staining in cardiomyocytes of heart muscle (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression mainly in skeletal and heart muscle. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min for nuclear MLIP (UniProt Q5VWP3; page retrieval rule).
Section 2
What Is the Expected MLIP Staining Pattern?
In paraffin-section IHC, expect mainly cytoplasmic MLIP staining in cardiomyocytes and skeletal myocytes, with weaker staining possible in smooth muscle cells (HPA tissue IHC: High, High and Medium, respectively). MLIP is also annotated at the nucleus, nuclear envelope and sarcolemma; it has no transmembrane segment (UniProt Q5VWP3). HPA rates the tissue IHC profile Approved, with medium consistency between staining and RNA data and external verification pending (HPA tissue IHC).
| Strong cytoplasmic staining in cardiomyocytes or skeletal myocytes, with weaker staining in smooth muscle cells. | This matches the reported IHC distribution: High in heart and skeletal muscle cells, Medium in smooth muscle cells, and mainly cytoplasmic overall (HPA tissue IHC). Assess intensity in the identified cells rather than averaging the whole section (general IHC practice). |
| Staining is confined to extracellular material, a lumen or granular deposits, without a convincing cellular pattern. | These locations do not match the reported predominantly cytoplasmic muscle-cell pattern (HPA tissue IHC). Inspect the section and detection controls for deposits or background before assigning the signal to MLIP (general IHC practice). Nuclear or membrane-associated signal alone should not be dismissed: those locations have separate support (UniProt Q5VWP3; HPA subcellular ICC-IF). |
| Strong staining appears in adipocytes or other cells reported as undetected, while muscle cells show little signal. | Adipocytes are reported as Not detected in adipose tissue and breast (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity; compare with an appropriate no-primary control and known-positive muscle section (general IHC practice; HPA tissue IHC). A low-level signal elsewhere is less decisive because several epithelial and neuronal cell types are reported Low (HPA tissue IHC). |
| Diffuse chromogen obscures cell boundaries or covers both expected positive and negative areas. | The distribution cannot be scored reliably against the reported muscle-cell pattern (HPA tissue IHC). Check the no-primary control and review blocking, washes and detection conditions as general IHC troubleshooting steps; this appearance alone does not establish an MLIP-specific cause (general IHC practice). |
| A heart or skeletal muscle section shows no convincing signal in its expected positive cells. | Those cells are reported High, so first confirm tissue identity and the assay controls (HPA tissue IHC; general IHC practice). Review the antibody's IHC-P instructions, detection reagents and retrieval settings if applicable (general IHC practice). The supplied sources do not establish MLIP-specific fixation or retrieval sensitivity. |
💡Expected MLIP appearanceCall a result positive when cytoplasmic staining is conspicuous in cardiomyocytes or skeletal myocytes, with possible weaker smooth-muscle staining; isolated extracellular deposits or diffuse cell-independent color are suspect (HPA tissue IHC; general IHC practice).
| Tissue and cell selection | Heart cardiomyocytes and skeletal myocytes offer reported High signal; smooth muscle cells are Medium (HPA tissue IHC). Adipocytes in adipose tissue are a reported Not detected comparison (HPA tissue IHC). Compare identified cell types, because mixed tissue can obscure which cells carry chromogen (general IHC practice). |
| Compartment used for scoring | HPA describes tissue staining as mainly cytoplasmic, while UniProt also annotates nucleus, nuclear envelope, PML body, cytosol and sarcolemma (HPA tissue IHC; UniProt Q5VWP3). Record compartment and cell type together; do not treat every departure from purely cytoplasmic staining as an artifact. |
| Topology and processing | MLIP has no transmembrane segment, signal peptide or propeptide, and its annotated chain spans residues 1–993 (UniProt Q5VWP3). Sarcolemma localization therefore does not by itself imply a membrane-spanning staining pattern (UniProt Q5VWP3 topology and localization). No tissue IHC effect of processing is established by these annotations. |
| Isoforms and epitope coverage | Four isoforms are listed (UniProt Q5VWP3). If a stain differs between tissues, check whether the antibody's documented epitope is shared across the isoforms before attributing the difference to expression (general IHC practice; UniProt Q5VWP3 isoforms). The supplied record gives no epitope map or isoform-specific tissue staining. |
| Strength of IHC evidence | HPA calls its tissue IHC profile Approved but notes medium staining–RNA consistency and pending external verification (HPA tissue IHC). HPA029252 has IHC Approved status (HPA antibodies). Use the reported pattern as a benchmark while interpreting unexpected staining with controls, rather than treating approval as proof of every signal's specificity (general IHC practice). |
| IF/ICC Q&A: should its pattern match tissue IHC exactly? | No exact match is established: HPA reports approved nucleoplasm and plasma-membrane localization, plus vesicles, in ICC-IF; tissue IHC is mainly cytoplasmic (HPA subcellular ICC-IF; HPA tissue IHC). UniProt also annotates nuclear and sarcolemma locations (UniProt Q5VWP3). Interpret each application against its own evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in cardiomyocytes or skeletal myocytes. | A failed staining run is possible; these cells are reported High, but the sources give no MLIP-specific fixation-effect finding (HPA tissue IHC). | Confirm section identity and assay controls, then check the antibody's IHC-P instructions, detection reagents and retrieval settings where applicable (general IHC practice). |
| Strong color appears in adipocytes. | This conflicts with reported Not detected adipocytes and could reflect cross-reactivity or endogenous detection activity (HPA tissue IHC; general IHC practice). | Compare a no-primary control and a known-positive muscle section; localize color to specific cells before scoring it as MLIP (general IHC practice; HPA tissue IHC). |
| Background is widespread and cell boundaries are unclear. | Diffuse color prevents comparison with the mainly cytoplasmic muscle-cell profile; the image alone cannot identify its cause (HPA tissue IHC; general IHC practice). | Review the no-primary control, blocking, washes and chromogen development, then reassess the cellular pattern (general IHC practice). |
| The only apparent positive signal is extracellular or particulate. | That appearance does not fit the reported muscle-cell staining and may represent a deposit or nonspecific signal (HPA tissue IHC; general IHC practice). | Inspect the deposits and detection controls; require a reproducible cellular pattern in an expected positive tissue before calling it MLIP (general IHC practice; HPA tissue IHC). |
| Nuclear or membrane-associated color accompanies muscle-cell staining. | UniProt lists nuclear, nuclear-envelope and sarcolemma locations, while HPA tissue IHC emphasizes cytoplasm (UniProt Q5VWP3; HPA tissue IHC). | Document the separate compartments and compare their cell distribution with controls; do not reject the stain solely because it is not exclusively cytoplasmic (general IHC practice; UniProt Q5VWP3). |
| Weak staining is seen outside heart and skeletal muscle. | Some nonmuscle cell types are reported Low, so weak staining outside the principal positive tissues is not automatically contradictory (HPA tissue IHC). | Identify the stained cell type and compare its reported HPA level; weigh weak signal against controls and the stronger expected muscle-cell pattern (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing MLIP staining
A12261 · IHCImmunohistochemistry of MLIP in human heart tissue with MLIP antibody at 5 μg/mL.
A12261 · IFImmunofluorescence of MLIP in human heart tissue with MLIP antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000146147-MLIP/tissue/cerebral+cortex#imid_19669843
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000146147-MLIP/tissue/colon#imid_19669761
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000146147-MLIP/tissue/liver#imid_19669762
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000146147-MLIP/tissue/kidney#imid_19669771
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA046654; green = MLIP, blue = nucleus, red = microtubules.
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA046654; green = MLIP, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MLIP IHC & IF
🧪Run heart muscle first; cardiomyocytes should stain strongly (HPA: High in cardiomyocytes). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); on the heart slide, assess non-cardiomyocyte areas for background without assuming every such cell is MLIP-negative (HPA: High in cardiomyocytes).
Positive control tissue: Heart muscle (Cardiomyocytes, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MLIP in Rh30, SK-MEL-30, U2OS, with annotated localisation: Nucleoplasm (approved), Plasma membrane (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and isotype controls, matching the isotype antibody to the catalog antibody’s host species and clonality, plus MLIP-knockout tissue as a biological negative if available (standard IHC practice). Quench endogenous peroxidase for chromogenic heart IHC and assess tissue autofluorescence if using heart sections for IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A12261 heart-tissue IHC caption does not state a fixative (selected-SKU caption). Retrieval dependency is unreported; optimize retrieval for paraffin sections rather than assuming a condition from the HPA staining pattern (HPA: High in cardiomyocytes; standard IHC practice). The supplied evidence does not establish whether frozen sections or IF are easier for MLIP; heart pigment can complicate interpretation, especially through autofluorescence in IF (standard IHC/IF practice).
HPA tissue IHC evidence for MLIP
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | High | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Medium | 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 → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MLIP IHC Tips
Troubleshoot MLIP staining in paraffin sections by checking retrieval, compartment and cell type before comparing chromogenic IHC results with IF.
How should I retrieve MLIP in paraffin sections when nuclear staining is weak?
Start with Tris-EDTA at pH 9.0, 95–98 °C for 20 min for chromogenic IHC on paraffin sections (page protocol). Let sections cool in retrieval buffer before washing, and keep heating and cooling conditions consistent across comparison slides (standard IHC practice). MLIP has reported nuclear and nuclear-envelope locations, while tissue IHC mainly shows cytoplasmic muscle staining; assess both compartments separately (UniProt Q5VWP3 localisation; HPA tissue IHC). If signal remains weak, compare a small retrieval time series while holding antibody concentration and detection constant, because excessive retrieval can damage tissue morphology (standard IHC practice).
Could fixation explain weak or uneven MLIP staining in my paraffin sections?
MLIP-specific sensitivity to fixation is unknown from the supplied evidence; do not infer it from tissue staining patterns, topology or phosphorylation sites (supplied evidence). The human heart image reports 5 µg/mL antibody but does not state its fixative (A12261 caption). For new paraffin specimens, record fixative, fixation duration and processing history, then compare matched sections under the same retrieval and detection conditions (standard IHC practice). Check whether weak regions also show poor morphology or uneven staining by a tissue control, and standardise fixation before interpreting a difference as altered MLIP expression (standard IHC practice).
Which MLIP staining compartments should I expect in heart and skeletal muscle?
In tissue IHC, expect predominantly cytoplasmic staining in heart and skeletal muscle; cardiomyocytes and skeletal myocytes are reported as High (HPA tissue IHC). MLIP is also annotated in the nucleus, nuclear envelope, PML bodies, cytosol and sarcolemma (UniProt Q5VWP3 localisation). Cell-based IF reports approved nucleoplasmic and plasma-membrane locations, with vesicles as an additional location; that evidence does not establish their relative prominence in tissue IHC (HPA subcellular; HPA tissue IHC). Score cytoplasmic, nuclear and membrane-associated signal separately, and verify boundaries with a counterstain and adjacent morphology before calling a compartment-specific change (standard IHC practice).
How can isoforms or epitope accessibility complicate MLIP IHC?
MLIP has 4 listed isoforms, but the supplied evidence does not map the catalog antibody’s epitope to any of them (UniProt Q5VWP3 isoforms; supplied evidence). Therefore, a negative section cannot establish that every MLIP isoform is absent; check the antibody’s documented immunogen or epitope before making an isoform-specific claim (standard IHC interpretation). Reported phosphoserines include positions 146 and 818, without evidence here that either changes this antibody’s binding (UniProt Q5VWP3 modified residues; supplied evidence). Compare retrieval conditions on matched sections and keep detection settings fixed when investigating epitope accessibility, while monitoring tissue integrity (standard IHC practice).
How can IF help assess an ambiguous MLIP IHC pattern?
Use IF as a separate cross-check of the cell and compartment assignment, since tissue IHC mainly reports cytoplasmic muscle staining and cell-based IF reports nucleoplasm and plasma membrane (HPA tissue IHC; HPA subcellular). Multiplex MLIP with a validated marker of the expected cardiomyocyte or myocyte population, plus a nuclear counterstain, to confirm which cells carry signal (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and check unstained tissue for autofluorescence, especially before interpreting faint signal (standard IF practice). For intracellular epitopes, evaluate controlled permeabilisation; for a surface-facing epitope, assess staining without permeabilisation, although this antibody’s epitope side is unreported (standard IF practice; supplied evidence).
What controls distinguish MLIP signal from chromogenic background?
Run a no-primary control through the same chromogenic workflow to reveal secondary-reagent and substrate background, and use an appropriate peroxidase block before DAB development (standard IHC practice). Compare background with a high-expression reference such as heart muscle cardiomyocytes or skeletal myocytes, while matching section thickness and development time (HPA tissue IHC; standard IHC practice). Adipocytes in adipose tissue are reported as Not detected, but this reference pattern is a comparison, not proof that every brown deposit is specific (HPA tissue IHC). If diffuse staining persists, optimise blocking, antibody concentration and washing one variable at a time; interpret DAB deposits against cellular boundaries and the no-primary slide (standard IHC practice).
How should I quantify MLIP staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored population before imaging, because cardiomyocytes and skeletal myocytes have reported High staining while smooth muscle cells are Medium (HPA tissue IHC). For cytoplasmic DAB, report an H-score from 0–300 or the percentage of positive target cells, using a fixed intensity threshold across slides (standard IHC practice). If counting discrete positive cells, report density per mm² of viable, analysable tissue and state the counting rule (standard IHC practice). Normalise to the number or area of the relevant muscle cells rather than total section area, exclude damaged regions consistently, and report nuclear or membrane-associated signal separately (standard IHC practice; UniProt Q5VWP3 localisation).
When is an unusual MLIP staining pattern likely to be artefactual?
A credible result should match the cell type and compartment being scored: tissue IHC mainly shows cytoplasmic signal in heart and skeletal muscle, with high staining in cardiomyocytes and myocytes (HPA tissue IHC). Nuclear or membrane-associated signal is biologically plausible, but requires independent compartment checks because those locations are documented separately from the dominant tissue IHC pattern (UniProt Q5VWP3 localisation; HPA subcellular; HPA tissue IHC). Treat staining confined to section edges, necrotic regions or the no-primary control as suspect, and examine residual endogenous peroxidase before attributing brown deposits to MLIP (standard IHC practice). Recheck unexpected positive cells against morphology and a matched control before concluding that MLIP distribution has changed (standard IHC practice).
Boster reagents
Best MLIP / Muscular LMNA-interacting protein IHC Antibodies
A12261 is a human-reactive anti-MLIP antibody with IHC and IF images from human heart tissue (catalog reactivity; A12261 image captions).
Real IHC data
Anti-MLIP Antibody
Cat # A12261
A12261 lists IHC-P and IF for human samples (catalog applications; catalog reactivity). Its IHC and IF images each show human heart tissue (A12261 image captions).
Which to pick: For tissue IHC, choose A12261 for human paraffin sections: IHC-P is listed, and its own IHC caption shows human heart tissue at 5 μg/mL; the fixative is unreported (catalog applications; A12261 IHC image caption). For IF, A12261 has a human-heart image at 20 μg/mL; ICC validation and clonality are unreported (catalog applications; A12261 IF image caption; catalog clone). No cross-species option is documented because A12261 lists human reactivity only (catalog reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q5VWP3 (MLIP_HUMAN, Muscular LMNA-interacting protein).
- Human Protein Atlas. MLIP tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MLIP subcellular location (ICC-IF): Mainly localized to the nucleoplasm and plasma membrane. In addition localized to vesicles..
- Human Protein Atlas. MLIP antibody validation summary (3 antibodies).
- Identification of a novel muscle A-type lamin-interacting protein (MLIP). The Journal of biological chemistry 2011 — PMC3103349.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MLIP (Muscular LMNA-interacting protein). Expected localisation Muscle cytoplasm (HPA tissue IHC); nuclear sites (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A12261 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Heart muscle (Cardiomyocytes); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A12261); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-MLIP, 5 μg/mL (datasheet A12261); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: IHC/RNA concordance is medium; verify with controls (HPA tissue IHC); Isoform / epitope: 4 isoforms; confirm antibody epitope coverage (UniProt); Regulation: Heart and skeletal muscle enriched (UniProt). Evidence: UniProt Q5VWP3, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm and plasma membrane. In addition…, Datasheet A12261, PMC3103349. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MLIP/immunohistochemistry-ihc-guide.