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LGALS2 / Galectin-2 · IHC design guide
Design Immunohistochemistry for LGALS2
Plan LGALS2 chromogenic IHC around cytoplasmic staining in gastrointestinal enterocytes and gallbladder glandular cells (HPA tissue IHC). The IHC-validated antibody A04114-2 has a recommended concentration of 2–5 μg/mL for paraffin sections (datasheet: A04114-2).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LGALS2 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 (HPA tissue IHC) | |
| Staining pattern | Enterocytes and glandular cells show cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04114-2) | |
| Positive control | Appendix+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 | Staining and RNA show only medium consistency (HPA tissue IHC) | |
| Regulation | Regulation not established (UniProt) | |
| Isoform / epitope | No annotated isoforms or processing; chain 1–132 (UniProt) |
Section 1
Recommended LGALS2 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 heat retrieval (datasheet A04114-2). One published LGALS2 IHC protocol uses citrate pH 6.0 retrieval on tumor microarrays (PMC9242595).
| Sample | Paraffin-embedded human pancreatic cancer tissue; fixative not specified (datasheet A04114-2) |
| Fixation | Image fixative and duration unreported (datasheet A04114-2); 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 A04114-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04114-2) |
| Primary antibody | Rabbit anti-LGALS2, 2-5μg/ml (datasheet A04114-2) |
| Primary incubation | Overnight at 4 °C (datasheet A04114-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A04114-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LGALS2-positive staining in enterocytes of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in gall bladder, gastrointestinal tract, exocrine pancreas and renal tubules. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 for the catalog antibody (datasheet A04114-2). Use citrate pH 6.0 to reproduce the published protocol (PMC9242595).
Section 2
What Is the Expected LGALS2 Staining Pattern?
In paraffin section IHC, expect mainly cytoplasmic LGALS2 staining in gastrointestinal enterocytes, gallbladder glandular cells and pancreatic exocrine glandular cells; kidney proximal tubules show a weaker reference pattern (HPA tissue IHC). HPA rates the tissue pattern Enhanced, with medium consistency between staining and RNA data (HPA tissue IHC). LGALS2 has no annotated transmembrane segment, and UniProt does not assign it a subcellular location (UniProt P05162 topology and subcellular annotation).
| Strong cytoplasmic staining in gastrointestinal enterocytes, gallbladder glands or pancreatic exocrine cells. | This matches HPA's High cell level observations and cytoplasmic tissue profile (HPA tissue IHC). Judge the cell type and compartment together: staining somewhere in a positive organ is insufficient if the expected cells remain clear. |
| Cytoplasmic staining of kidney proximal tubule cell bodies, less intense than the High reference cells. | This is compatible with HPA's Medium kidney observation (HPA tissue IHC). Compare matched sections and staining conditions before treating an intensity difference as biological; the HPA levels are reference categories, not a quantitative cutoff. |
| Predominantly nuclear, membranous or extracellular staining in an IHC positive reference tissue. | That does not match the reported cytoplasmic tissue IHC profile (HPA tissue IHC). Review morphology and controls before calling it LGALS2; HPA separately reports mitochondrial and additional nucleoplasmic localisation by ICC-IF, which is a different assay (HPA subcellular ICC-IF). |
| Strong signal in an unexpected cell population while the expected positive cells are faint or clear. | Consider antibody cross-reactivity or endogenous detection activity, and check the detection only control. HPA reports High staining in specified enterocytes and glandular cells, not every cell in those tissues (HPA tissue IHC). |
| Diffuse staining obscures cell borders, or a known High reference cell population has no signal. | The result cannot yet support a confident pattern call. Diffuse signal calls for background controls; absent signal calls for a run control and review of IHC processing. HPA's High reference populations provide a comparison, but do not diagnose the failed step (HPA tissue IHC). |
💡Expected LGALS2 appearanceCall the IHC result positive when cytoplasmic signal is clear in High reference enterocytes or gallbladder or pancreatic glandular cells, with Medium proximal tubule staining also compatible; isolated nuclear or widespread unrelated cell staining is suspect and needs controls (HPA tissue IHC; HPA subcellular ICC-IF for the assay distinction).
| Reference tissue and cell type | Appendix, colon, duodenum, rectum and small intestine enterocytes, gallbladder glandular cells and pancreatic exocrine glandular cells are High; kidney proximal tubules are Medium (HPA tissue IHC). Use the named cells when selecting and reading a positive reference. |
| Reported negative comparison | HPA reports Not detected in adipocytes of adipose tissue and respiratory epithelial cells of bronchus, among other specified populations (HPA tissue IHC). These are cell level comparisons; another cell type in the same section is not automatically a negative control. |
| Antibody validation and evidence limit | HPA lists IHC Enhanced for HPA003536 and CAB025198 and describes tissue reliability as Enhanced with medium staining–RNA consistency (HPA antibodies; HPA tissue IHC). This supports comparison with the reported pattern, but cannot establish the specificity of every signal in a new run. |
| Protein structure and processing | UniProt records a 132 aa chain, a galectin domain at residues 4–131, no transmembrane segment, no signal peptide or propeptide, and no annotated glycosylation sites (UniProt P05162). These entries do not identify an antibody epitope or establish antigen retrieval or fixation sensitivity. |
| IF/ICC localisation question | Does a mitochondrial pattern in IF/ICC contradict cytoplasmic IHC? HPA reports mitochondria as the main ICC-IF location and nucleoplasm as an additional location, while its tissue IHC profile is cytoplasmic (HPA subcellular ICC-IF; HPA tissue IHC). Interpret each against its own assay evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| No visible staining in a High reference cell population. | The IHC run may have lost detectable signal; the slide alone cannot identify whether processing, retrieval, antibody incubation or detection is responsible. | Check an on-run positive control and review the established paraffin IHC steps one at a time. Use HPA's High enterocyte or glandular cell references to assess recovery (HPA tissue IHC); no LGALS2-specific retrieval or fixation sensitivity is supplied. |
| Uniform haze or precipitate makes cytoplasmic detail hard to score. | Background from detection reagents, insufficient blocking or incomplete washing can obscure a cellular pattern in routine chromogenic IHC. | Inspect the detection only control and section morphology, then review blocking, washes and chromogen development. Score LGALS2 only after the expected cells can be distinguished from background (HPA tissue IHC for expected cells). |
| Strong staining appears in cells outside the reported positive populations. | Cross-reactivity or endogenous detection activity is possible; an unexpected cell signal is not proof of either cause. | Compare a detection only control and a matched positive reference. Check whether the named High cells show the expected cytoplasmic pattern before assigning LGALS2 positivity (HPA tissue IHC). |
| Kidney proximal tubules stain more weakly than gastrointestinal enterocytes. | HPA classifies proximal tubules as Medium and the specified enterocytes as High (HPA tissue IHC). | Read intensity relative to the correct cell level reference under comparable run conditions. Do not reject a clear proximal tubule cell body signal solely because it is weaker than a High reference (HPA tissue IHC). |
| A nominally negative section contains scattered stained cells. | HPA's Not detected calls apply to named cells, such as adipocytes in adipose tissue, rather than every cell in the section (HPA tissue IHC). | Identify the stained cell type first. If the named negative population itself stains, inspect controls for background or cross-reactivity before treating the result as LGALS2 (HPA tissue IHC). |
| IF/ICC shows mitochondrial puncta or additional nucleoplasmic signal. | Those locations are reported for ICC-IF, whereas the supplied tissue IHC profile is cytoplasmic (HPA subcellular ICC-IF; HPA tissue IHC). | Use the separate IF/ICC guide for that assay. For this paraffin IHC interpretation, compare the slide with HPA's cytoplasmic pattern in the specified tissue cells (HPA tissue IHC). |
Experiment evidence
IHC & IF images showing LGALS2 staining
A04114-2 · IHCIHC analysis of Galectin 2/LGALS2 using anti-Galectin 2/LGALS2 antibody (A04114-2). Galectin 2/LGALS2 was detected in paraffin-embedded sect
A04114-2 · IHCIHC analysis of Galectin 2/LGALS2 using anti-Galectin 2/LGALS2 antibody (A04114-2). Galectin 2/LGALS2 was detected in paraffin-embedded sect
A04114-2 · IFIF analysis of Galectin 2/LGALS2 using anti-Galectin 2/LGALS2 antibody (A04114-2). Galectin 2/LGALS2 was detected in immunocytochemical sect
A04114 · IHCImmunohistochemistry analysis of paraffin-embedded human heart tissue, using LEG2 Antibody. The picture on the right is blocked with the syn
A04114 · IFImmunofluorescence analysis of COS7 cells, using LEG2 Antibody. The picture on the right is blocked with the synthesized peptide.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000100079-LGALS2/tissue/cerebral+cortex#imid_1273916
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000100079-LGALS2/tissue/colon#imid_1263266
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000100079-LGALS2/tissue/liver#imid_1263259
HPA · GallbladderHuman Protein Atlas IHC, Gallbladder. Source: https://www.proteinatlas.org/ENSG00000100079-LGALS2/tissue/gallbladder#imid_1263235
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003536; green = LGALS2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003536; green = LGALS2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LGALS2 IHC & IF
🧪Run colon first and score enterocytes for staining (HPA: High in colon enterocytes). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes), and compare enterocytes with adjacent non-enterocyte stromal cells on the colon slide as an internal background reference.
Positive control tissue: Appendix (Enterocytes, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LGALS2 in A-431, CACO-2, U2OS, with annotated localisation: Mitochondria (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control appropriate to the primary antibody’s clonality (selected-SKU caption: rabbit primary). Use LGALS2 knockout material as a biological negative where available, and block endogenous peroxidase before chromogenic detection in colon sections (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU tissue-IHC caption does not state a fixative. The selected-SKU caption demonstrates heat-mediated retrieval in EDTA at pH 8.0 for a paraffin section of pancreatic cancer tissue; it does not establish that the same retrieval conditions are required for colon (selected-SKU caption). The supplied evidence does not establish whether frozen sections or IF are easier for LGALS2; assess intestinal luminal staining and endogenous peroxidase as possible sources of chromogenic background when scoring colon (standard IHC practice).
HPA tissue IHC evidence for LGALS2
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 |
|---|---|---|---|---|
| Appendix | Enterocytes | High | Protein (IHC) | HPA → |
| Colon | Enterocytes | High | Protein (IHC) | HPA → |
| Duodenum | Enterocytes | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Pancreas | Exocrine 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 → |
| 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 → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LGALS2 IHC Tips
Troubleshoot LGALS2 staining in paraffin sections by checking retrieval, controls, cell identity and compartment before assigning a biological interpretation.
Which retrieval conditions should I use when LGALS2 staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A04114-2). The documented tissue image used this retrieval before an overnight 2 µg/mL primary incubation at 4°C (datasheet A04114-2). If staining remains weak, check that sections were fully deparaffinised, then vary heating duration on adjacent sections while keeping the EDTA buffer and detection conditions constant (standard IHC practice). Compare each condition with a high-expression reference, such as gallbladder glandular cells or intestinal enterocytes (HPA tissue IHC). Reject a hotter condition if morphology deteriorates or background rises, even when the chromogenic signal increases (standard IHC practice).
Could fixation explain a weak or uneven LGALS2 stain?
Target-specific fixation sensitivity is unknown because the selected tissue-IHC caption does not state a fixative (datasheet A04114-2). Record the fixative, fixation duration and processing history for each block before comparing staining intensity across specimens (standard IHC practice). Review a reference section from the same processing batch and check for uneven staining across thick areas, folds or tissue edges (standard IHC practice). If the batch is suspect, compare adjacent sections using the documented EDTA pH 8.0 retrieval and 2 µg/mL overnight primary incubation (datasheet A04114-2). Neither the tissue-expression profile nor the protein’s topology establishes how LGALS2 responds to fixation (HPA tissue IHC; UniProt P05162).
Where should convincing LGALS2 staining appear in tissue sections?
Expect predominantly cytoplasmic staining in the documented gallbladder, gastrointestinal, exocrine pancreatic and renal tubular tissue patterns (HPA tissue IHC). Enterocytes and gallbladder glandular cells provide strongly stained cell populations for comparison, while kidney proximal tubule cell bodies show a medium level (HPA tissue IHC). Score the cell type and compartment separately on a hematoxylin-counterstained section; diffuse chromogen over luminal debris should not be assigned to intact cells (standard IHC practice). LGALS2 has no annotated transmembrane segment or signal peptide, so a crisp membrane-only pattern needs independent confirmation (UniProt P05162). Cell-line ICC/IF reports mitochondrial and additional nucleoplasmic localisation, which should be evaluated separately from tissue IHC (HPA subcellular).
How do I assess a staining pattern when the antibody epitope is unspecified?
Check the catalog antibody’s stated immunogen or epitope before attributing an unexpected staining pattern to LGALS2 (standard IHC practice). The reference record lists a 132-residue chain, a galectin domain spanning residues 4–131, and no annotated isoforms, glycosylation sites or modified residues (UniProt P05162). Those annotations do not identify the antibody’s binding site or prove that fixation preserves it (UniProt P05162; standard IHC practice). Compare the suspect section with a tissue reference stained in the same run under EDTA pH 8.0 retrieval (datasheet A04114-2; HPA tissue IHC). If an independent antibody is available, agreement in cell type and compartment strengthens an epitope-specific interpretation (standard IHC practice).
How can IF help check an ambiguous LGALS2 IHC pattern?
Use IF/ICC as a separate confirmation experiment, since the selected antibody’s supplied image documents chromogenic tissue IHC (datasheet A04114-2). Multiplex LGALS2 with an independently validated marker for the cell type being examined, such as an enterocyte marker when evaluating intestinal epithelium (HPA tissue IHC; standard IF practice). Choose fluorophores after imaging an unstained section so tissue autofluorescence does not mask the LGALS2 channel, and include single-label controls (standard IF practice). For an intracellular epitope, optimise permeabilisation to permit access to the expected compartment without disrupting morphology; LGALS2 has no annotated transmembrane segment (UniProt P05162; standard IF practice). Assess mitochondrial or nucleoplasmic overlap separately from the tissue-IHC cytoplasmic pattern (HPA subcellular; HPA tissue IHC).
What should I check when DAB background obscures LGALS2 staining?
First inspect a no-primary control alongside the stained section to identify signal arising from detection reagents or endogenous tissue activity (standard IHC practice). The documented workflow uses 10% goat serum, a biotinylated secondary and streptavidin-biotin detection with DAB (datasheet A04114-2). Check peroxidase blocking, secondary-only staining and potential endogenous biotin signal before changing the primary concentration (standard IHC practice). If background persists, titrate around the documented 2 µg/mL primary concentration and keep retrieval at EDTA pH 8.0 for the comparison (datasheet A04114-2; standard IHC practice). Interpret improved contrast against a reference with expected cellular staining, rather than by background reduction alone (HPA tissue IHC; standard IHC practice).
How should I quantify LGALS2 across differently composed tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring; the reported tissue pattern is cytoplasmic and varies by cell type (HPA tissue IHC). For a defined population, record the percentage of positive cells and staining intensity, then calculate an H-score using percentages at intensity levels 0–3 (standard IHC practice). Alternatively, report positive-cell density per mm² of viable annotated tissue when cell counts better fit the specimen (standard IHC practice). Normalise comparisons to the number or area of the relevant cells, rather than total section area when tissue composition differs (standard IHC practice). Apply one threshold and imaging setup across the run, and exclude folds, edges and necrotic regions by prespecified rules (standard IHC practice).
How can I distinguish true LGALS2 staining from artefact?
A credible positive should label intact cells in a plausible cytoplasmic pattern and reproduce in an appropriate reference tissue (HPA tissue IHC; standard IHC practice). Strong intestinal enterocytes, gallbladder glandular cells or pancreatic exocrine cells offer documented comparisons; adipocytes and adrenal glandular cells are reported as undetected (HPA tissue IHC). Treat staining confined to necrosis, section edges or luminal debris as suspect until it survives repeat staining and histological review (standard IHC practice). A membrane-only pattern warrants scrutiny because LGALS2 has no annotated transmembrane segment, while cell-line IF reports mitochondrial and additional nucleoplasmic signal (UniProt P05162; HPA subcellular). Check no-primary and peroxidase controls before calling diffuse brown deposits LGALS2 (standard IHC practice).
Boster reagents
Best LGALS2 / Galectin-2 IHC Antibodies
Both antibodies list IHC and IF applications with human reactivity (catalog applications/reactivity). A04114 also lists mouse and rat reactivity (A04114 reactivity); their figures show paraffin-section IHC and cell IF (image captions).
Real IHC data
Anti-Galectin 2/LGALS2 Antibody ®
Cat # A04114-2
Real IHC data
Anti-LEG2 LGALS2 Antibody
Cat # A04114
A04114-2 lists human IHC and IF applications; its figures show IHC in human pancreatic cancer paraffin sections and IF in A549 cells (A04114-2 applications/reactivity and image captions). A04114 lists human, mouse and rat reactivity with IHC and IF applications; its figures show human heart paraffin-section IHC and COS7 cell IF, each with a peptide-blocked comparison (A04114 applications/reactivity and image captions).
Which to pick: For tissue IHC, start with A04114-2 when its documented workflow fits: the human pancreatic cancer paraffin-section caption specifies EDTA pH 8 retrieval and 2 μg/ml primary antibody; A04114 instead shows human heart paraffin-section staining with peptide blocking, and neither IHC caption reports the fixative (respective IHC captions). For human cell IF/ICC, A04114-2 has an A549 IF figure using 5 μg/ml primary antibody, while A04114 has a COS7 IF figure with peptide blocking (respective IF captions). For planned mouse or rat work, choose the rabbit polyclonal A04114 based on its listed reactivity, then verify staining in the intended sample because its supplied IHC figure is human heart (A04114 host, dilution_raw, reactivity and IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P05162 (LEG2_HUMAN, Galectin-2).
- Human Protein Atlas. LGALS2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. LGALS2 subcellular location (ICC-IF): Mainly localized to the mitochondria. In addition localized to the nucleoplasm..
- Human Protein Atlas. LGALS2 antibody validation summary (2 antibodies).
- In vivo multidimensional CRISPR screens identify Lgals2 as an immunotherapy target in triple-negative breast cancer. Science advances 2022 — PMC9242595.
- LGALS2 suppresses the progression of papillary thyroid carcinoma by regulating the PI3K/AKT pathway. Gland surgery 2022 — PMC9547712.
- PubMed PMID:1375225 — UniProt-cited evidence.
- PubMed PMID:15461802 — UniProt-cited evidence.
- PubMed PMID:10591208 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LGALS2 (Galectin-2). Expected localisation Cytoplasmic staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04114-2). Validated IHC antibody: A04114-2 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Enterocytes); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human pancreatic cancer tissue; fixative not specified (datasheet A04114-2); 2. Fixation: Image fixative and duration unreported (datasheet A04114-2); 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 A04114-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04114-2); 8. Primary antibody: Rabbit anti-LGALS2, 2-5μg/ml (datasheet A04114-2); 9. Primary incubation: Overnight at 4 °C (datasheet A04114-2). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA show only medium consistency (HPA tissue IHC); Isoform / epitope: No annotated isoforms or processing; chain 1–132 (UniProt); Regulation: Regulation not established (UniProt). Evidence: UniProt P05162, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the mitochondria. In addition localized to the…, Datasheet A04114-2, PMC9242595. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LGALS2/immunohistochemistry-ihc-guide.