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LUC7L3 / Luc7-like protein 3 · IHC design guide
Design Immunohistochemistry for LUC7L3
Plan paraffin-section LUC7L3 staining around its widespread nuclear tissue pattern (HPA tissue IHC). This guide covers fixation consistency, nuclear scoring, and epitope coverage across its 2 isoforms (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LUC7L3 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 | Nuclear staining (HPA tissue IHC); speckle localization (UniProt) | |
| Staining pattern | Widespread nuclear staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A08633-3) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Cisplatin can alter subnuclear localization (UniProt) | |
| Regulation | Broad expression; regulation unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; confirm epitope coverage for both (UniProt) |
Section 1
Recommended LUC7L3 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A08633-3) is accompanied by one published LUC7L3 cell-block protocol (PMC12530123).
| Sample | Paraffin-embedded human highly differentiated adenocarcinoma of the colon tissue; fixative not specified (datasheet A08633-3) |
| Fixation | Image fixative and duration unreported (datasheet A08633-3); 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 A08633-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08633-3) |
| Primary antibody | Rabbit anti-LUC7L3, 1-2 μg/ml (datasheet A08633-3) |
| Primary incubation | Overnight at 4 °C (datasheet A08633-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A08633-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LUC7L3-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet A08633-3); the published protocol does not specify retrieval (PMC12530123).
Section 2
What Is the Expected LUC7L3 Staining Pattern?
LUC7L3 should appear in nuclei across many tissue cell types (HPA: ubiquitous nuclear expression; Enhanced tissue IHC reliability). High staining is reported in adipocytes, glandular cells, hematopoietic cells, respiratory epithelial cells and selected brain cells (HPA: tissue IHC). Nuclear speckles are its more precise location (UniProt O95232: nucleus speckle); it has no transmembrane segment (UniProt O95232: topology).
| Nuclear chromogen appears in several cell populations, including the cell types reported as High in the chosen tissue (HPA: tissue IHC). | This fits the broad nuclear pattern (HPA: ubiquitous nuclear expression). Compare intensity within identifiable cells rather than requiring every nucleus to stain equally (general IHC interpretation); HPA's High calls describe observed populations, not a mandatory intensity threshold for every section. |
| Strong cytoplasmic or membrane staining dominates while nuclei are weak or clear. | That compartment conflicts with the nuclear tissue profile (HPA: ubiquitous nuclear expression) and the annotated nuclear speckle location (UniProt O95232: subcellular location). Review morphology and controls before calling it LUC7L3; a broad extranuclear deposit can reflect nonspecific staining or detection artefact (general IHC practice). |
| Signal is restricted to cells outside the population expected from the selected HPA tissue image. | Check cell identification first: LUC7L3 is widely expressed (UniProt O95232: tissue specificity), so an additional positive cell type alone does not prove cross-reactivity. If staining follows pigment, blood, or reagent-associated activity rather than nuclei, evaluate cross-reactivity and endogenous detection activity (general IHC practice). |
| A uniform haze obscures nuclei and tissue boundaries. | This is difficult to score as the reported nuclear pattern (HPA: tissue IHC profile). Review a reagent-only control, blocking, washes and chromogen development to locate nonspecific binding or detection background (general IHC practice); do not infer a target-specific fixation effect from this appearance. |
| No nuclei stain in an HPA-reported High population, such as cerebellar granular-layer cells (HPA: High). | Treat the result as technically unresolved before concluding that LUC7L3 is absent. Check section integrity, antibody and detection performance, and a matched positive control (general IHC practice). An HPA High observation supports a useful comparator, but does not guarantee staining in every specimen or run (HPA: tissue IHC). |
💡Expected LUC7L3 appearanceCall the IHC result positive when identifiable cells show convincing nuclear chromogen, especially in an HPA-reported High population (HPA: ubiquitous nuclear expression; tissue IHC); predominantly cytoplasmic colour or uniform tissue haze is suspect (UniProt O95232: nucleus speckle; general IHC interpretation).
| Tissue and cell context | Expression is widespread (UniProt O95232: tissue specificity), and HPA reports High nuclear staining in several named cell populations (HPA: tissue IHC); use the chosen tissue's observed pattern as a comparator, not an exclusivity rule. |
| Subnuclear resolution | Nuclear speckles are supported by ICC-IF (HPA: enhanced nuclear speckles) and UniProt (O95232: nucleus speckle). In chromogenic sections, assess a nuclear result; apparent speckling depends on optical and tissue resolution (general IHC practice). |
| Antibody evidence | The listed HPA antibodies have Enhanced IHC status (HPA: HPA018475, HPA018484, HPA020017). This supports the reported pattern; it does not validate an unlisted antibody, its dilution, or a new tissue preparation (HPA: antibody validation summary). |
| Treatment context | UniProt reports that cisplatin affects subnuclear localization (UniProt O95232: subcellular-location note). Document treatment when comparing nuclear detail; the record supplies no direction, magnitude, or IHC-specific prediction for the change. |
| Molecular forms and target access | Two isoforms are listed (UniProt O95232: isoforms), with no signal peptide, propeptide, or transmembrane segment annotated (UniProt O95232: processing and topology). The payload gives no epitope map, so isoform detection and retrieval needs remain antibody dependent. |
| Target-specific fixation evidence | No fixation sensitivity is reported in the supplied UniProt or HPA records. Retrieval and section processing may be optimized as general IHC practice, but these records do not support a LUC7L3-specific fixation or epitope-masking claim. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive comparator lacks nuclear staining. | The staining run or section may have failed; absence cannot be assigned to LUC7L3 from one slide (general IHC practice). | Check tissue integrity and positive-control nuclei, then verify antibody application and detection steps against the established IHC workflow (general IHC practice); compare with the selected HPA High cell population (HPA: tissue IHC). |
| Cytoplasmic colour overwhelms nuclear colour. | The result conflicts with the expected compartment (HPA: ubiquitous nuclear expression; UniProt O95232: nucleus speckle); nonspecific binding or chromogen background is possible (general IHC practice). | Compare with a reagent-only control and inspect whether colour tracks tissue edges or deposits; reassess blocking, washes and development (general IHC practice). |
| Unexpected cells stain while the HPA-listed cells remain clear. | Cell identity, variable biology, cross-reactivity, or endogenous detection activity may explain the mismatch (general IHC interpretation); HPA High calls are observations, not exclusive expression rules (HPA: tissue IHC). | Verify cell identity on the counterstained section, compare another HPA-listed positive population, and use appropriate detection controls (general IHC practice). |
| All compartments show faint colour. | Diffuse background prevents a reliable nuclear call (HPA: nuclear tissue profile); excess detection signal or incomplete washing may contribute (general IHC practice). | Review reagent-only control, wash performance and development endpoint; score only cell-associated nuclear colour above local background (general IHC practice). |
| Nuclear detail differs between treated and untreated samples. | Cisplatin can affect subnuclear localization (UniProt O95232: subcellular-location note), while section resolution can alter its appearance (general IHC practice). | Record treatment and compare matched sections using the same scoring approach; avoid assigning a specific redistribution without supporting images or controls (general IHC interpretation). |
| Q: What should an IF/ICC image show? | The subcellular reference is mainly nuclear speckles (HPA: ICC-IF, enhanced nuclear speckles; UniProt O95232: nucleus speckle). | A: Look for nuclear speckle enrichment and compare with an appropriate nuclear counterstain (HPA: ICC-IF; general IF interpretation); use the separate IF/ICC guide for method details. |
Experiment evidence
IHC & IF images showing LUC7L3 staining
A08633-3 · IHCIHC analysis of CROP/LUC7L3 using anti-CROP/LUC7L3 antibody (A08633-3). CROP/LUC7L3 was detected in a paraffin-embedded section of human hig
A08633-3 · IHCIHC analysis of CROP/LUC7L3 using anti-CROP/LUC7L3 antibody (A08633-3). CROP/LUC7L3 was detected in a paraffin-embedded section of human lar
A08633-3 · IFIF analysis of CROP/LUC7L3 using anti-CROP/LUC7L3 antibody (A08633-3) and anti-Beta Tubulin antibody (M01857-3). CROP/LUC7L3 was detected in
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000108848-LUC7L3/tissue/cerebral+cortex#imid_5318856
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000108848-LUC7L3/tissue/colon#imid_5318798
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000108848-LUC7L3/tissue/liver#imid_5318783
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000108848-LUC7L3/tissue/kidney#imid_5318800
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018475; green = LUC7L3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018475; green = LUC7L3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LUC7L3 IHC & IF
🧪Run bone marrow first: hematopoietic cells should stain (HPA: High in bone marrow hematopoietic cells). HPA detects LUC7L3 in all 45 scored tissues, so use no-primary and isotype controls for the negative; mature erythrocytes on the positive slide should lack nuclear staining because they have no nuclei (HPA: no negative tissue; standard IHC practice).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: LUC7L3 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LUC7L3 in A-431, U-251MG, U2OS, U2OS, siRNA 1 (10x), U2OS, siRNA 1 (40x), U2OS, scrambled (10x), U2OS, scrambled (40x), with annotated localisation: Nuclear speckles (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section and a section incubated with host- and isotype-matched rabbit IgG; use a validated LUC7L3 knockout specimen as a biological negative if available (caption: rabbit primary antibody; standard IHC practice). Quench endogenous peroxidase in bone marrow before HRP/DAB detection, particularly in myeloid cells (caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A08633-3 paraffin-section caption does not state the fixative (selected-SKU tissue-IHC caption). That caption uses heat retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required; bone marrow endogenous peroxidase can complicate HRP/DAB interpretation (selected-SKU tissue-IHC caption; standard IHC practice). The supplied evidence does not establish whether frozen sections or IF/ICC are easier; HPA ICC-IF images provide a nuclear-speckle localization reference (HPA: subcellular localization).
HPA tissue IHC evidence for LUC7L3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: LUC7L3 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced LUC7L3 IHC Tips
Use nuclear staining as the main readout for LUC7L3 in paraffin section IHC, with the catalog antibody’s documented conditions as the starting point.
Which retrieval condition should I try first for LUC7L3 paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet A08633-3). The catalog antibody’s paraffin-section image used that condition, followed by 10% goat serum blocking and 2 μg/mL primary antibody overnight at 4°C (caption A08633-3). Compare retrieved and unretrieved serial sections while holding antibody concentration, detection and development constant, and judge success by nuclear staining with preserved tissue structure (standard IHC practice; HPA: ubiquitous nuclear expression). If staining remains weak, test a different retrieval buffer or heating schedule on adjacent sections as a fallback, recording whether background and morphology change (standard IHC practice).
How can I assess whether fixation is masking the LUC7L3 epitope?
Target-specific fixation sensitivity is unknown: the catalog image identifies a paraffin-embedded section but does not state its fixative (caption A08633-3). Record the fixative, fixation duration and processing history for each block, then compare sections with similar morphology under the same EDTA pH 8.0 retrieval and detection conditions (datasheet A08633-3; standard IHC practice). Include a previously staining control section in the same run so a weak result can be assessed against run performance (standard IHC practice). Interpret a fixation effect only if matched material shows a reproducible difference after other processing variables have been controlled (standard IHC practice).
What staining pattern should count as correctly localised LUC7L3?
Score staining within nuclei first, because tissue IHC shows ubiquitous nuclear expression and subcellular evidence places LUC7L3 mainly in nuclear speckles (HPA: tissue IHC; HPA: nuclear speckles). Chromogenic sections may show nuclear signal without resolving individual speckles, so compare stained nuclei with the counterstain and local tissue architecture (standard IHC practice). Treat predominantly membrane or diffuse extracellular DAB as a specificity concern; LUC7L3 has no annotated transmembrane segment (UniProt O95232 topology; standard IHC practice). If comparing treated samples, document treatment alongside staining, since cisplatin can alter its subnuclear localisation (UniProt O95232 subcellular annotation).
Can this stain distinguish LUC7L3 isoforms or changes near the antibody epitope?
Do not assign staining to one isoform without an epitope map or isoform-specific validation: the record lists 2 isoforms, while the supplied caption does not identify the antibody epitope (UniProt O95232 isoforms; caption A08633-3). The record also lists modified residues, including phosphoserines at positions 110 and 115, but supplies no evidence that those modifications affect this antibody (UniProt O95232 modified residues). If isoform identity matters, compare the epitope sequence with each isoform and validate the interpretation independently (standard IHC practice). Report the observed nuclear staining as LUC7L3 immunoreactivity until those checks support a narrower claim (HPA: ubiquitous nuclear expression; standard IHC practice).
How should I check LUC7L3 localisation in a multiplex IF comparison?
On the separate IF/ICC workflow, pair LUC7L3 with an identity marker for the cell population being compared and a nuclear counterstain; adipocytes are among the populations with high tissue IHC staining (HPA: high in adipocytes; standard IF practice). Choose spectrally separated fluorophores, considering a far-red channel if tissue autofluorescence obscures shorter-wavelength signal (standard IF practice). Because the expected signal is nuclear and LUC7L3 has no transmembrane segment, assess permeabilisation sufficient to admit antibody to nuclei after fixation (HPA: nuclear speckles; UniProt O95232 topology; standard IF practice). Check each channel separately before interpreting overlap, and do not treat the paraffin-section caption as IF fixation evidence (caption A08633-3; standard IF practice).
How do I distinguish LUC7L3 staining from chromogenic background?
Inspect a no-primary control and the tissue edges alongside the test section; diffuse DAB, edge accentuation or staining over damaged tissue warrants caution (standard IHC practice). The documented image used 10% goat serum blocking, 2 μg/mL primary antibody overnight at 4°C, and an HRP/DAB detection system (caption A08633-3). Check that peroxidase blocking and washes are effective, and shorten DAB development if the control accumulates colour (standard IHC practice). Compare nuclear signal with the counterstain, because the expected tissue pattern is nuclear rather than a uniform coating across the section (HPA: ubiquitous nuclear expression; standard IHC practice).
What should I measure when comparing LUC7L3 IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define positive nuclei against a same-run control, then report the percentage of positive nuclei and, when intensity matters, a nuclear H-score (standard IHC practice; HPA: ubiquitous nuclear expression). Calculate the H-score from the percentages at intensity grades 0–3, giving a possible range of 0–300 (standard IHC practice). Normalise counts to evaluable nuclei within the same annotated cell compartment, and use cells per mm² only when cell density is itself the question (standard IHC practice). Keep retrieval, exposure to DAB and scoring thresholds consistent across sections, since the supplied image gives an example condition rather than a quantitative reference scale (caption A08633-3; standard IHC practice).
What evidence would make a LUC7L3-positive IHC result convincing?
Look for nuclear staining in intact cells that exceeds same-run no-primary background and agrees with the expected ubiquitous nuclear tissue pattern (HPA: tissue IHC; standard IHC practice). High staining in adipocytes or hematopoietic cells is plausible, but neither cell population alone proves antibody specificity (HPA: high in adipocytes and hematopoietic cells; standard IHC practice). Exclude section edges, necrotic areas and endogenous peroxidase colour before scoring, and question staining confined to membranes or extracellular material (standard IHC practice; UniProt O95232 topology). Report treatment context when comparing subnuclear patterns because cisplatin can affect LUC7L3 localisation, without inferring an expression change from redistribution alone (UniProt O95232 subcellular annotation; standard IHC practice).
Boster reagents
Best LUC7L3 / Luc7-like protein 3 IHC Antibodies
A08633-3 has paraffin-section IHC images from human tissues and an IF/ICC image from U2OS cells (catalog image captions). Its listed reactivity covers human, mouse, and rat (catalog: reactivity).
Real IHC data
Anti-CROP/LUC7L3 Antibody ®
Cat # A08633-3
A08633-3 is listed for IHC and has images from human paraffin sections of colon adenocarcinoma, colonic large B-cell lymphoma, liver cancer, and lung cancer (catalog: applications; IHC image captions). A08633-3 is also listed for IF/ICC, with an IF image from U2OS cells (catalog: applications; IF image caption).
Which to pick: For tissue IHC, choose A08633-3: its images document human paraffin sections with EDTA pH 8.0 retrieval and 2 μg/ml primary antibody; the fixative is unreported (catalog: IHC image captions). For IF/ICC, the same SKU has an image at 5 μg/ml in U2OS cells (catalog: IF image caption). For mouse or rat samples, A08633-3 lists reactivity with both species, although the supplied IHC and IF images show human samples (catalog: reactivity; image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O95232 (LC7L3_HUMAN, Luc7-like protein 3).
- Human Protein Atlas. LUC7L3 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. LUC7L3 subcellular location (ICC-IF): Mainly localized to the nuclear speckles..
- Human Protein Atlas. LUC7L3 antibody validation summary (3 antibodies).
- The molecular chaperone TRAP1 promotes translation of Luc7I3 mRNA to enhance ovarian cancer cell proliferation. RNA (New York, N.Y.) 2025 — PMC12530123.
- PubMed PMID:10631324 — UniProt-cited evidence.
- PubMed PMID:16462885 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LUC7L3 (Luc7-like protein 3). Expected localisation Nuclear staining (HPA tissue IHC); speckle localization (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A08633-3). Validated IHC antibody: A08633-3 at 1-2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human highly differentiated adenocarcinoma of the colon tissue; fixative not specified…; 2. Fixation: Image fixative and duration unreported (datasheet A08633-3); 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 A08633-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08633-3); 8. Primary antibody: Rabbit anti-LUC7L3, 1-2 μg/ml (datasheet A08633-3); 9. Primary incubation: Overnight at 4 °C (datasheet A08633-3). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Cisplatin can alter subnuclear localization (UniProt); Isoform / epitope: 2 isoforms; confirm epitope coverage for both (UniProt); Regulation: Broad expression; regulation unreported (UniProt). Evidence: UniProt O95232, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nuclear speckles., Datasheet A08633-3, PMC12530123. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LUC7L3/immunohistochemistry-ihc-guide.