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LPIN1 / Phosphatidate phosphatase LPIN1 · IHC design guide
Design Immunohistochemistry for LPIN1
Plan LPIN1 IHC-P around cytoplasmic staining in most tissues, with high signal in kidney tubules, lung macrophages and Leydig cells (HPA tissue IHC). This guide covers fixation consistency, controls, chromogenic detection and cell-specific scoring.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LPIN1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic signal; high in lung macrophages and kidney tubules (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Kidney+4 more · see all | |
| Negative control | Cerebellum+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02467) | |
| Caveat | Staining has medium consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Muscle enriched; adipose abundant (UniProt) | |
| Isoform / epitope | 7 isoforms; no extracellular region; check epitope coverage (UniProt) |
Section 1
Recommended LPIN1 IHC & IF Protocols
The catalog antibody IHC-P protocol is accompanied by published LPIN1 IHC conditions for colonic tissue (PMC12395820) and breast cancer tissue (PMC8124251).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A02467); 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-LPIN1, 1:100–1:500 starting range (standard) |
| 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 | LPIN1-positive staining in cells in tubules of kidney (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page retrieval); the colonic tissue protocol uses EDTA at 95 °C for 10 min (PMC12395820).
Section 2
What Is the Expected LPIN1 Staining Pattern?
LPIN1 should usually appear cytoplasmic in tissue sections (HPA tissue IHC: cytoplasmic in most tissues), with strong staining in kidney tubular cells, lung macrophages and testis Leydig cells (HPA tissue IHC: High). HPA rates the tissue pattern Approved, with medium consistency between staining and RNA data (HPA tissue IHC). Cytosolic and membrane-associated localisation is plausible for a protein without a transmembrane segment (UniProt Q14693: localisation and topology).
| Distinct cytoplasmic staining in kidney tubules, lung macrophages or testis Leydig cells. | This matches the reported High cell-specific tissue pattern (HPA tissue IHC). Compare cells within the section; staining of the entire tissue is not the expected readout (HPA tissue IHC: cell-level observations). |
| Predominantly nuclear interior staining, with little cytoplasmic signal. | Treat a nuclear-only pattern as suspect: tissue IHC is mainly cytoplasmic (HPA tissue IHC), although UniProt lists the nuclear membrane (UniProt Q14693: localisation). Check whether any apparent nuclear signal follows the rim before calling it discordant. |
| Strong staining in cells reported as undetected, such as pancreatic exocrine glandular cells. | This conflicts with that cell-level reference (HPA tissue IHC: Not detected). Cross-reactivity or endogenous detection activity is possible (standard IHC practice); confirm cell identity and inspect controls before assigning LPIN1. |
| Uniform colour across stroma, empty spaces and several unrelated cell types. | This is more consistent with background than the reported cell-specific pattern (HPA tissue IHC). Review the no-primary control, blocking and detection steps (standard chromogenic IHC practice). |
| No staining in kidney tubular cells or lung macrophages. | A blank known-positive cell population suggests an assay problem, though HPA reports only medium staining–RNA consistency (HPA tissue IHC). Check section quality, controls and the antibody's documented IHC use before interpreting the absence biologically (standard IHC practice). |
💡Expected LPIN1 appearanceCall a section positive when cytoplasmic signal is clearly enriched in kidney tubular cells, lung macrophages or testis Leydig cells at the reported High level (HPA tissue IHC); uniform background or isolated nuclear-interior colour is a suspect pattern (HPA tissue IHC: cytoplasmic profile; standard IHC practice).
| Cell and tissue context | HPA reports High staining in kidney tubules, lung macrophages and Leydig cells, but Medium in adipocytes (HPA tissue IHC). UniProt describes skeletal muscle specificity and abundant adipose expression (UniProt Q14693); RNA or tissue-level descriptions do not replace cell-level IHC observations. |
| Compartment and movement | LPIN1 is listed in cytosol, ER membrane and nuclear membrane, and can move from cytosol to ER after KAT5 acetylation (UniProt Q14693: localisation). Interpret variable cytoplasmic distribution cautiously; these annotations do not predict a particular paraffin-section pattern. |
| Antibody validation by application | HPA038021 is IHC Approved, while HPA024591 is ICC Approved (HPA antibodies). These are distinct application records; the ICC result does not establish an IHC pattern for HPA024591. HPA's tissue profile has medium staining–RNA consistency (HPA tissue IHC). |
| Isoforms and antibody target | UniProt lists 7 LPIN1 isoforms (UniProt Q14693). The supplied records give no epitope or isoform coverage for the IHC antibody, so a difference between sections cannot be assigned to a particular isoform from staining alone. |
| Protein topology and processing | LPIN1 has no transmembrane segment, signal peptide or propeptide in the supplied record (UniProt Q14693: topology and processing). Membrane-associated localisation therefore does not establish integral membrane staining or extracellular deposition; evaluate the observed cellular compartment instead (UniProt Q14693: localisation). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive tissue is blank. | The assay may have failed; kidney tubular cells and lung macrophages are reported High (HPA tissue IHC). | Check the tissue and run controls, then review the IHC antibody's documented conditions and detection reagents (standard IHC practice). No LPIN1-specific retrieval or fixation effect is supplied. |
| Colour is diffuse across the section. | Non-specific binding, residual endogenous detection activity or incomplete washing can produce background (standard chromogenic IHC practice). | Inspect the no-primary control and review blocking, washes and detection chemistry (standard IHC practice). Score only cell-associated staining against the HPA cell-level reference (HPA tissue IHC). |
| Nuclear-interior staining dominates. | A nuclear-only result differs from the usual cytoplasmic tissue profile (HPA tissue IHC); the nuclear membrane annotation alone does not explain filled nuclei (UniProt Q14693: localisation). | Compare cytoplasm and nuclear rim in known-positive cells, then check controls and antibody specificity before accepting the pattern (standard IHC practice). |
| An expected negative cell type stains strongly. | Cross-reactivity or endogenous chromogen activity may mimic signal (standard IHC practice); pancreatic exocrine glandular cells are reported Not detected (HPA tissue IHC). | Verify the cell type and compare positive tissue with no-primary and detection controls (standard IHC practice). Treat the HPA negative as a reference observation, not proof of universal absence. |
| Only a low-expression tissue looks blank. | Glial cells in hippocampus and caudate are reported Low, whereas several other cell types are Not detected (HPA tissue IHC). A faint or absent result there is less informative than a blank High reference. | Assess kidney tubular cells or lung macrophages in the same run as positive references (HPA tissue IHC); use controls to judge assay performance (standard IHC practice). |
| IF/ICC shows peripheral signal: is that an IHC positive pattern? | ICC-IF reports approved cytosolic localisation plus additional plasma membrane localisation (HPA subcellular); tissue IHC reports a generally cytoplasmic profile (HPA tissue IHC). | Interpret the IF/ICC observation in its own application context and assess paraffin IHC against the tissue cell pattern (HPA subcellular; HPA tissue IHC). The two application records do not establish identical staining distributions. |
Experiment evidence
IHC & IF images showing LPIN1 staining
A02467 · IHCImmunohistochemistry of LPIN1 in human small intestine tissue with LPIN1 antibody at 5 μg/ml.
A02467 · IFImmunofluorescence of LPIN1 in human small intestine tissue with LPIN1 antibody at 20 μg/ml.
A02467-2 · IFIF analysis of Lipin 1/LPIN1 using anti-Lipin 1/LPIN1 antibody (A02467-2). Lipin 1/LPIN1 was detected in an immunocytochemical section of PC
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000134324-LPIN1/tissue/cerebral+cortex#imid_9866052
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000134324-LPIN1/tissue/colon#imid_9863718
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000134324-LPIN1/tissue/liver#imid_9863711
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000134324-LPIN1/tissue/kidney#imid_9863720
HPA · ICC-IF ASC52teloHuman Protein Atlas immunofluorescence, ASC52telo, antibody HPA024591; green = LPIN1, blue = nucleus, red = microtubules.
HPA · ICC-IF ASC52teloHuman Protein Atlas immunofluorescence, ASC52telo, antibody HPA024591; green = LPIN1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LPIN1 IHC & IF
🧪Run kidney first: tubular cells should stain strongly (HPA: High in cells in tubules). Use cerebellum as a negative tissue, focusing on granular-layer cells (HPA: Not detected in cells in granular layer); on the kidney slide, assess adjacent non-tubular areas for background, without assuming those cells are LPIN1-negative (HPA: High designation applies to cells in tubules).
Positive control tissue: Kidney (Cells in tubules, HPA High)
Negative control tissue: Cerebellum (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LPIN1 in ASC52telo, SK-MEL-30, U2OS, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an irrelevant antibody matched to the primary antibody’s host species and, for a monoclonal antibody, its isotype (standard IHC practice). Confirm specificity with LPIN1-knockout material or an antigen-peptide competition control where available (standard IHC practice).
For kidney, block endogenous biotin if using avidin–biotin detection, because renal tubules can produce biotin-related background (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected A02467 small-intestine IHC caption does not state its fixative (caption: fixative not stated). Antigen-retrieval dependency is also unreported; optimize retrieval empirically for paraffin IHC, and do not assume frozen sections or IF will be easier (standard IHC practice). With kidney as the positive tissue, distinguish tubular signal from endogenous-biotin background when using avidin–biotin detection (HPA: High in cells in tubules; standard IHC practice).
HPA tissue IHC evidence for LPIN1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium 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 |
|---|---|---|---|---|
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Lung | Macrophages | High | Protein (IHC) | HPA → |
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LPIN1 IHC Tips
Troubleshoot LPIN1 staining in paraffin sections by checking retrieval, tissue controls, subcellular pattern, and cell-specific scoring before interpreting signal.
How should I adjust retrieval when LPIN1 staining is weak or uneven?
Use Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page retrieval setting). Keep section thickness, cooling time, and detection conditions constant while comparing retrieved sections with a no-primary control (standard IHC practice). Examine intact tissue away from section edges before increasing retrieval time, because edge staining and tissue damage can distort the comparison (standard IHC practice). If staining remains weak, test citrate pH 6.0 on matched sections as a fallback optimization, documenting the change rather than treating it as a validated LPIN1 condition (standard IHC practice). Compare cytoplasmic signal in renal tubule cells with the negative control (HPA: High in kidney tubule cells).
Could fixation explain weak or patchy LPIN1 staining in paraffin sections?
LPIN1-specific fixation sensitivity is unknown from the supplied evidence, so record fixative, fixation duration, and processing history for each paraffin block (supplied LPIN1 evidence: no target-specific fixation comparison; standard IHC practice). The A02467 small-intestine image caption reports antibody use at 5 µg/ml but does not state its fixative (A02467 tissue-IHC caption). Compare adjacent sections from blocks with known processing histories under the same Tris-EDTA pH 9.0 retrieval and detection conditions (page retrieval setting; standard IHC practice). Include an independently stained positive control and a no-primary control in each run to distinguish a block-specific loss of signal from assay failure (standard IHC practice).
Where should LPIN1 staining appear, and how should unexpected nuclear signal be checked?
Expect predominantly cytoplasmic staining in tissue sections (HPA: cytoplasmic expression in most tissues), consistent with cytosolic LPIN1 (UniProt Q14693: cytosol). LPIN1 is also annotated at the endoplasmic reticulum membrane and nuclear membrane, and cytosol-to-endoplasmic-reticulum translocation follows KAT5 acetylation (UniProt Q14693: subcellular location). Thus, record perinuclear or membrane-adjacent signal separately from diffuse nuclear chromogen; localisation alone does not establish the protein's functional state (UniProt Q14693: subcellular location; standard IHC interpretation). Compare the pattern in high-staining renal tubule cells with a no-primary section and examine individual cells at higher magnification (HPA: High in kidney tubule cells; standard IHC practice).
Can this antibody distinguish LPIN1 isoforms or modified forms in tissue?
LPIN1 has 7 annotated isoforms and multiple modified residues, including phosphoserines at positions 106 and 150 (UniProt Q14693: isoforms and modified residues). The supplied A02467 caption gives a tissue staining concentration but no epitope sequence or isoform-specific validation (A02467 tissue-IHC caption). Check the antibody's documented immunogen and isoform coverage before assigning staining to one splice form or phosphorylation state (standard IHC interpretation). If epitope information remains unavailable, report the result as LPIN1 immunoreactivity and compare matched tissue sections under identical retrieval and detection conditions (standard IHC practice). A changed compartmental pattern alone cannot identify an isoform (UniProt Q14693: multiple subcellular locations and isoforms).
How can IF test whether LPIN1 signal belongs to the expected cells?
On the separate IF/ICC guide, multiplex LPIN1 with a validated marker for the cell population being examined, such as a macrophage marker when assessing lung macrophages (HPA: High in lung macrophages; standard IF practice). Choose spectrally separated fluorophores and inspect an unstained section to select a detection channel with less tissue autofluorescence (standard IF practice). Use mild permeabilisation for an intracellular epitope, then compare permeabilised and untreated samples if access is uncertain; LPIN1 has no transmembrane segment and is annotated in the cytosol and at the endoplasmic reticulum membrane (UniProt Q14693: topology and subcellular location; standard IF practice). Keep single-stain and no-primary controls for bleed-through and nonspecific fluorescence (standard IF practice).
What should I check when LPIN1 chromogen covers many unrelated cells?
First inspect a no-primary control and a matched section stained with the catalog antibody to separate detection background from primary-dependent signal (standard IHC practice). In peroxidase-based chromogenic IHC, check the peroxidase block before DAB development and shorten DAB exposure if diffuse brown deposit obscures cell boundaries (standard IHC practice). Review blocking, washes, and antibody concentration together; the A02467 small-intestine caption used 5 µg/ml, which is an image condition rather than a universal optimum (A02467 tissue-IHC caption; standard IHC practice). Assess cytoplasmic staining in expected positive cells alongside HPA-listed unstained cell populations, while treating HPA's Approved rating as medium consistency with RNA data (HPA: tissue IHC).
How should I score LPIN1 IHC across sections with different cell mixtures? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic compartment before scoring, because HPA reports cytoplasmic expression across many tissues and different levels among cell types (HPA: tissue IHC profile). For a uniform population, report the percentage of positive cells and a prespecified intensity-based H-score; keep the intensity scale and threshold fixed across slides (standard IHC quantification). For scattered positive cells, report positive cells per mm² of viable tissue and the proportion of the relevant cell population that is positive (standard IHC quantification). Normalise comparisons to viable tissue area or the counted target-cell population, and exclude folds, edges, and necrotic regions using the same rules for every section (standard IHC practice).
When is LPIN1 staining convincing rather than a tissue or detection artefact?
A convincing result shows reproducible, mainly cytoplasmic signal in intact expected cells, such as renal tubule cells or lung macrophages (HPA: cytoplasmic profile; HPA: High in kidney tubule cells and lung macrophages). Check the same run's no-primary control for chromogen generated by the detection system, including residual endogenous peroxidase (standard chromogenic IHC practice). Treat isolated edge deposits, necrotic areas, and staining that obscures cell boundaries as suspect until they recur in well-preserved central tissue (standard IHC interpretation). A predominantly diffuse nuclear pattern warrants further control testing because cytosol is the principal annotated location, although LPIN1 is also annotated at the nuclear membrane (HPA: cytosol approved; UniProt Q14693: nuclear membrane).
Boster reagents
Best LPIN1 / Phosphatidate phosphatase LPIN1 IHC Antibodies
A02467 has human small-intestine IHC and IF images (catalog: image captions); A02467-2 has human PC-3 ICC/IF data (catalog: IF image caption).
Real IHC data
Anti-LPIN1 Antibody
Cat # A02467
Real IF data
Anti-Lipin 1/LPIN1 Antibody ®
Cat # A02467-2
A02467 will render with a human small-intestine IHC image; its listed applications also include IF (catalog: IHC image caption; applications). A02467-2 will render with a human PC-3 ICC/IF image (catalog: IF image caption).
Which to pick: Choose A02467 for tissue IHC: it lists IHC-P and shows staining in human small intestine (catalog: applications; IHC image caption); the caption does not report the fixative (catalog: IHC image caption). Choose A02467-2 for human cell ICC/IF: its PC-3 image used 5 μg/mL (catalog: applications; IF image caption). For planned mouse or rat work, A02467 lists both species and IHC-P, while its reported IHC and IF images are from human tissue (catalog: reactivity; applications; image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q14693 (LPIN1_HUMAN, Phosphatidate phosphatase LPIN1).
- Human Protein Atlas. LPIN1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. LPIN1 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the plasma membrane..
- Human Protein Atlas. LPIN1 antibody validation summary (2 antibodies).
- Significant correlation between ferroptosis-associated genes and ulcerative colitis: implications for diagnosis and treatment. European journal of medical research 2025 — PMC12395820.
- IL-33-Induced Transcriptional Activation of LPIN1 Accelerates Breast Tumorigenesis. Cancers 2021 — PMC8124251.
- Lipin 1 deficiency causes adult-onset myasthenia with motor neuron dysfunction in humans and neuromuscular junction defects in zebrafish. Theranostics 2021 — PMC7806489.
- PubMed PMID:8724849 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LPIN1 (Phosphatidate phosphatase LPIN1). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A02467 at 1:100–1:500 (standard starting range) (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in tubules); negative Cerebellum. 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 A02467); 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-LPIN1, 1:100–1:500 starting range (standard); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02467); Caveat: Staining has medium consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 7 isoforms ; no extracellular region; check epitope coverage (UniProt); Regulation: Muscle enriched; adipose abundant (UniProt). Evidence: UniProt Q14693, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the plasma…, Datasheet A02467, PMC12395820, PMC8124251. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LPIN1/immunohistochemistry-ihc-guide.