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LYPLA1 / Acyl-protein thioesterase 1 · IHC design guide
Design Immunohistochemistry for LYPLA1
Plan LYPLA1 staining in paraffin sections using cytoplasmic glandular staining as a reference pattern (HPA tissue IHC). Compare the catalog antibody’s IHC conditions with positive and negative tissue controls (datasheet A07249-2; HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LYPLA1 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 | Predominantly cytoplasmic in tissue (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in glandular cells of several tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A07249-2) | |
| Positive control | Colon+4 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 | Esophageal carcinoma stains despite negative normal squamous cells (datasheet A07249-2; HPA tissue IHC) | |
| Regulation | No specific expression regulator reported (UniProt) | |
| Isoform / epitope | 2 isoforms; check antibody epitope coverage (UniProt) |
Section 1
Recommended LYPLA1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published LYPLA1 staining protocols for head and neck biopsies, kidney sections, and breast cancer tissue microarrays (PMC7367650; PMC9676193; PMC10463390).
| Sample | Paraffin-embedded human esophageal squamous carcinoma tissue; fixative not specified (datasheet A07249-2) |
| Fixation | Image fixative and duration unreported (datasheet A07249-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A07249-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A07249-2) |
| Primary antibody | Rabbit anti-LYPLA1, 2-5 μg/ml (datasheet A07249-2) |
| Primary incubation | Overnight at 4 °C (datasheet A07249-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A07249-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LYPLA1-positive staining in glandular cells of colon (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 for the catalog antibody (datasheet: A07249-2); published protocols used Tris-EDTA or citrate retrieval (PMC7367650; PMC9676193).
Section 2
What Is the Expected LYPLA1 Staining Pattern?
LYPLA1 should appear mainly in the cytoplasm, with weak cell, nuclear and endoplasmic reticulum membrane localization possible (UniProt O75608 localization). It has no transmembrane segment (UniProt O75608 topology). In paraffin sections, look first at glandular cells of the colon, duodenum, gallbladder, rectum, small intestine or stomach, where staining is high (HPA: tissue IHC). HPA calls the tissue staining reliability Enhanced but reports only medium consistency with RNA expression (HPA: tissue IHC).
| Glandular cells show clear cytoplasmic chromogen, especially in colon, duodenum, gallbladder, rectum, small intestine or stomach (HPA: High in glandular cells). | This fits the reported tissue pattern and predominant cytoplasmic localization (HPA: tissue IHC; UniProt O75608 localization). Judge intensity within the identified cell type; a generally dark section alone does not establish specific staining (general IHC practice). |
| Strong, isolated nuclear or sharply membrane-only staining dominates, with little glandular cytoplasm stained. | That distribution conflicts with the predominant cytoplasmic pattern (UniProt O75608 localization; HPA: tissue IHC). Weak nuclear or cell membrane signal remains plausible (UniProt O75608 localization), so check controls and antibody behavior before calling the dominant pattern an artefact (general IHC practice). |
| Adipocytes or esophageal squamous epithelial cells stain prominently while glandular cells do not. | Those cell types were not detected in the HPA tissue survey, whereas several glandular cell populations were high (HPA: tissue IHC). The mismatch warrants checks for cross-reactivity or endogenous chromogenic activity; neither cause is established by HPA alone (general IHC practice). |
| Chromogen is spread across the section, including spaces between cells or the negative control. | A diffuse deposit cannot be scored as LYPLA1's reported cellular pattern (HPA: cytoplasmic tissue expression). Review blocking, washes, detection reagents and the negative control for background sources (general IHC practice); the record does not identify a LYPLA1-specific cause. |
| A well-preserved glandular area in a reported high-staining tissue has no detectable signal. | This disagrees with the reported HPA pattern for that cell type (HPA: High in glandular cells). First confirm cell identity and that a control section developed; then review retrieval and detection conditions as general IHC troubleshooting, without assuming LYPLA1-specific fixation sensitivity (general IHC practice). |
💡Expected LYPLA1 appearanceCall a section positive when glandular-cell cytoplasm is clearly stained, with high signal expected in the listed gastrointestinal and gallbladder tissues (HPA: tissue IHC); dominant isolated nuclear staining or equally strong staining in HPA-undetected cell types needs investigation (UniProt O75608 localization; HPA: tissue IHC).
| Compartment and topology | Predominantly cytoplasmic localization guides scoring; weak cell, nuclear and endoplasmic reticulum membrane signal can occur (UniProt O75608 localization). Absence of a transmembrane segment does not rule out the reported weak membrane association (UniProt O75608 topology and localization). |
| Cell type within a tissue | HPA reports high staining in glandular cells at six named sites, medium in adrenal gland and appendix glandular cells, and undetected staining in selected adipocytes, squamous cells and other cell types (HPA: tissue IHC). Score the stated cell population rather than treating an entire organ as uniformly positive or negative. |
| Strength of supporting evidence | HPA050941 has Enhanced IHC validation, while the tissue profile has Enhanced reliability with medium antibody–RNA consistency (HPA: antibody validation; HPA: tissue IHC). These labels support the observed pattern but do not validate every antibody, section or unexpected compartment. |
| Isoforms and processing | UniProt lists two LYPLA1 isoforms and a chain spanning residues 1–230, with no signal peptide or propeptide reported (UniProt O75608 isoforms and processing). The supplied sources do not map the IHC antibody epitope or show whether isoforms stain differently; do not infer isoform identity from the slide. |
| Situation | Likely cause | Next action |
|---|---|---|
| High-staining glandular cells are blank (HPA: tissue IHC). | The expected cellular signal is missing; the supplied sources do not establish a LYPLA1-specific fixation or retrieval effect. | Verify cell identity and a working positive control, then review retrieval, primary antibody and chromogen steps (general IHC practice). |
| The section has widespread faint brown haze. | Diffuse signal lacks the reported cytoplasmic cell pattern (HPA: tissue IHC); background or endogenous detection activity is possible (general IHC practice). | Compare a primary-omission control and review blocking, washes and detection chemistry (general IHC practice). |
| Only nuclei are strongly stained. | Predominantly cytoplasmic localization is expected; HPA's uncertain nucleoplasmic IF finding does not establish a nuclear-dominant IHC pattern (UniProt O75608 localization; HPA: subcellular ICC-IF). | Check the negative control and repeat scoring against glandular cytoplasm; investigate persistent nuclear-dominant signal (general IHC practice). |
| Adipocytes or esophageal squamous cells stain more than nearby glandular cells. | HPA reports those particular cell types as not detected, while several glandular populations are high (HPA: tissue IHC). | Confirm cell identification, inspect controls for nonspecific or endogenous signal, and avoid calling those cells LYPLA1-positive on intensity alone (general IHC practice). |
| A weak membrane rim is visible alongside cytoplasm. | Weak cell and nuclear membrane localization is reported, despite no transmembrane segment (UniProt O75608 localization and topology). | Score the accompanying cytoplasmic pattern and cell type; investigate a membrane-only result with controls (HPA: tissue IHC; general IHC practice). |
| What should IF/ICC show? | HPA reports supported cytosol and uncertain nucleoplasm in ICC-IF, with a caution that the finding uses antibodies targeting proteins from multiple genes (HPA: subcellular ICC-IF). | Expect cytosolic signal as the better-supported comparison; treat nucleoplasmic signal cautiously (HPA: subcellular ICC-IF). Use the separate IF/ICC guide for experimental conditions. |
Experiment evidence
IHC & IF images showing LYPLA1 staining
A07249-2 · IHCIHC analysis of LYPLA1 using anti-LYPLA1 antibody (A07249-2). LYPLA1 was detected in a paraffin-embedded section of human esophageal squamou
A07249-2 · IHCIHC analysis of LYPLA1 using anti-LYPLA1 antibody (A07249-2). LYPLA1 was detected in a paraffin-embedded section of human rectum adenocarcin
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000120992-LYPLA1/tissue/cerebral+cortex#imid_14530688
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000120992-LYPLA1/tissue/colon#imid_14530614
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000120992-LYPLA1/tissue/liver#imid_14530631
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000120992-LYPLA1/tissue/kidney#imid_14530632
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA063151; green = LYPLA1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA063151; green = LYPLA1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LYPLA1 IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use esophageal squamous epithelium as a negative tissue comparator (HPA: Not detected in esophageal squamous epithelial cells); on the colon slide, independently verified LYPLA1-negative cells should show only counterstain and background, since HPA does not identify an internal negative cell type there (HPA: High in colon glandular cells).
Positive control tissue: Colon (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LYPLA1 in A-431, SH-SY5Y, with annotated localisation: Nucleoplasm (uncertain), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host and format, and a LYPLA1 knockout sample or validated peptide-block control (standard IHC practice; caption: rabbit anti-LYPLA1 A07249-2). For colon sections, quench endogenous peroxidase and inspect background before interpreting HRP-DAB staining (caption: peroxidase secondary and DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A07249-2 paraffin-section caption does not state the fixative (caption: fixative not stated). The reported IHC procedure uses heat-mediated retrieval in EDTA at pH 8.0, but the supplied evidence does not establish whether retrieval is required (caption: EDTA retrieval, pH 8.0). Frozen sections cannot be judged easier from these sources; IF/ICC images exist for A-431 and SH-SY5Y, while intestinal tissue background remains a practical staining check (HPA subcellular: ICC-IF images; standard IHC practice).
HPA tissue IHC evidence for LYPLA1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Rectum | Glandular cells | High | Protein (IHC) | HPA → |
| Small intestine | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LYPLA1 IHC Tips
Troubleshoot LYPLA1 staining in paraffin sections by checking retrieval, cellular compartment, controls and scoring before interpreting chromogenic signal.
Which retrieval condition should I start with for LYPLA1 in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A07249-2). The selected tissue image used this condition before incubation with 2 μg/ml antibody overnight at 4°C, so keep those conditions together during an initial comparison (datasheet A07249-2). If staining is weak, vary heating duration on adjacent sections while holding antibody concentration, detection and development constant; include a well-stained reference section in every run (standard IHC practice). Judge improvement by clearer cytoplasmic staining with preserved morphology, since cytoplasm is the predominant reported compartment for LYPLA1 (UniProt O75608 localization; HPA: cytoplasmic expression in several tissues).
How can I assess whether fixation is causing weak LYPLA1 staining?
Target-specific sensitivity to fixation is unknown from the supplied evidence, and the selected paraffin-section caption does not report a fixative (datasheet A07249-2: fixative unreported). Compare sections with documented processing histories, holding the EDTA pH 8.0 retrieval and staining run constant (datasheet A07249-2; standard IHC practice). Record fixation type and duration, section age, morphology and the fraction of evaluable cells before attributing weak signal to antigen loss (standard IHC practice). A processing effect is more credible if similarly handled reference tissue also changes, but staining patterns alone cannot establish how LYPLA1 responds to fixation (standard IHC practice).
What staining compartment should I expect, and how should I assess membrane signal?
Expect predominantly cytoplasmic staining; weak cell, nuclear-membrane and endoplasmic-reticulum localization is also reported for LYPLA1 (UniProt O75608 localization). HPA tissue IHC likewise describes cytoplasmic expression across several tissues, while its ICC/IF record supports cytosol and labels nucleoplasm uncertain (HPA tissue IHC; HPA subcellular). Score cytoplasmic intensity separately from any crisp membrane or nuclear signal, using a counterstain and comparable fields to confirm cellular boundaries (standard IHC practice). Because LYPLA1 has no annotated transmembrane segment, diffuse cytoplasmic signal should not be rejected merely because membrane staining is faint (UniProt O75608 topology and localization).
Can this IHC result distinguish LYPLA1 isoforms or an epitope affected by processing?
LYPLA1 has 2 listed isoforms, but the supplied antibody caption does not identify its epitope or establish isoform selectivity (UniProt O75608 isoforms; datasheet A07249-2). Treat staining as LYPLA1-associated signal without assigning it to a specific isoform, and obtain the immunogen sequence before making an isoform claim (standard antibody-validation practice). UniProt lists no signal peptide, propeptide or transmembrane segment and notes an N6-acetyllysine at residue 224; none establishes whether this antibody's binding changes with processing or acetylation (UniProt O75608 processing, topology and modified residues). If results conflict with another reagent, compare documented epitopes and independent controls before inferring a biological isoform difference (standard IHC practice).
How should IF help resolve ambiguous LYPLA1 staining in these IHC sections?
Use the separate IF/ICC guide for the fluorescence workflow; its images include A-431 and SH-SY5Y cells, with cytosol supported and nucleoplasm uncertain (HPA subcellular). For a tissue comparison, multiplex LYPLA1 with a validated marker of the cell type being assessed, then examine overlap at the single-cell level rather than comparing adjacent regions alone (standard IF practice). Choose spectrally separated fluorophores and an acquisition channel with low tissue autofluorescence, checking unstained material before interpreting dim signal (standard IF practice). Match permeabilisation to antibody epitope access: an intracellular epitope needs access through the plasma membrane, while epitope location for the catalog antibody is not supplied (standard IF practice; datasheet A07249-2).
How can I separate LYPLA1 staining from diffuse DAB background?
The selected paraffin-section image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and peroxidase-based DAB detection (datasheet A07249-2). Include a no-primary control, apply a peroxidase block, and keep DAB development time consistent when comparing sections; these are general chromogenic IHC controls (standard IHC practice). If background persists, compare antibody concentration and wash conditions on adjacent sections while retaining EDTA pH 8.0 retrieval (datasheet A07249-2; standard IHC practice). A convincing result should resolve into cell-associated cytoplasmic staining rather than uniform color over tissue and empty spaces (UniProt O75608 localization; standard IHC practice).
How should I quantify LYPLA1 IHC across tissues with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and analysis area before scoring, because HPA reports high staining in glandular cells of colon, duodenum, gallbladder, rectum, small intestine and stomach (HPA tissue IHC). For those cells, record percent positive and cytoplasmic intensity, then calculate an H-score as the sum of each intensity category multiplied by its percentage of cells (standard IHC practice). Use the same thresholds, illumination and DAB development across sections, and exclude folds, edges and necrotic regions from the denominator (standard IHC practice). Normalize to evaluable cells of the specified type or report positive-cell density per mm² when tissue composition varies (standard IHC practice).
What evidence makes a LYPLA1-positive IHC field biologically credible?
A credible positive field has reproducible, cell-associated cytoplasmic staining, consistent with the predominant localization reported by UniProt and HPA (UniProt O75608 localization; HPA tissue IHC). In glandular tissue, compare the defined cells with HPA's high-staining glandular examples; do not assume every cell in a section shares that profile (HPA tissue IHC). Recheck isolated nuclear-only signal, stained section edges, necrotic areas and color in the no-primary control before calling a field positive (HPA subcellular: nucleoplasm uncertain; standard IHC practice). Endogenous peroxidase can also produce chromogenic signal, so assess the peroxidase-blocked control alongside morphology and the expected compartment (standard IHC practice).
Boster reagents
Best LYPLA1 / Acyl-protein thioesterase 1 IHC Antibodies
A07249-2 has IHC images from human paraffin sections (catalog IHC captions); M07249-1 lists ICC/IF for human, mouse and rat, with no IF image supplied (catalog applications, reactivity and IF image alts).
Real IHC data
Anti-LYPLA1 Antibody ®
Cat # A07249-2
A07249-2 lists IHC and shows LYPLA1 staining in paraffin sections of human esophageal squamous carcinoma, rectum adenocarcinoma and lung cancer (catalog applications and IHC captions). M07249-1 lists ICC/IF and human, mouse and rat reactivity, but supplies no IF image (catalog applications, reactivity and IF image alts).
Which to pick: Choose A07249-2 for tissue IHC: its own captions document human paraffin sections, EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody; the fixative is unreported (catalog A07249-2 IHC captions). Choose M07249-1 for IF/ICC based on its listed application, rabbit monoclonal clone 21L68 and 1:50 IF dilution; no IF image is supplied (catalog M07249-1 applications, clone, dilution and IF image alts). For cross-species work, both list human, mouse and rat reactivity, but the supplied IHC images show human sections only and no IF images are supplied (catalog reactivity and image alts).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75608 (LYPA1_HUMAN, Acyl-protein thioesterase 1).
- Human Protein Atlas. LYPLA1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. LYPLA1 subcellular location (ICC-IF): Localized to the nucleoplasm and cytosol. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. LYPLA1 antibody validation summary (2 antibodies).
- Lipid and protein tumor markers for head and neck squamous cell carcinoma identified by imaging mass spectrometry. Oncotarget 2020 — PMC7367650.
- Renal lysophospholipase A1 contributes to Enterococcus faecalis-induced hypertension by enhancing sodium reabsorption. iScience 2022 — PMC9676193.
- Identifying prognostic markers in spatially heterogeneous breast cancer microenvironment. Journal of translational medicine 2023 — PMC10463390.
- PubMed PMID:11080636 — UniProt-cited evidence.
- PubMed PMID:11042152 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LYPLA1 (Acyl-protein thioesterase 1). Expected localisation Predominantly cytoplasmic in tissue (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A07249-2). Validated IHC antibody: A07249-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Colon (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human esophageal squamous carcinoma tissue; fixative not specified (datasheet A07249-2); 2. Fixation: Image fixative and duration unreported (datasheet A07249-2); 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: EDTA pH 8.0 (datasheet A07249-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A07249-2); 8. Primary antibody: Rabbit anti-LYPLA1, 2-5 μg/ml (datasheet A07249-2); 9. Primary incubation: Overnight at 4 °C (datasheet A07249-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Esophageal carcinoma stains despite negative normal squamous cells (datasheet A07249-2; HPA tissue IHC); Isoform / epitope: 2 isoforms; check antibody epitope coverage (UniProt); Regulation: No specific expression regulator reported (UniProt). Evidence: UniProt O75608, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm and cytosol. Caution: Based on antibodies targeting…, Datasheet A07249-2, PMC7367650, PMC9676193. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LYPLA1/immunohistochemistry-ihc-guide.