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LGALS2 / Galectin-2 · IHC design guide
Design Immunohistochemistry for LGALS2
HPA reports high LGALS2 staining in intestinal enterocytes and gallbladder and pancreatic glandular cells (HPA tissue IHC). Use this guide to plan paraffin IHC controls and interpret cytoplasmic staining.
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 in GI, gallbladder, pancreatic and renal cells (HPA tissue IHC) | |
| Staining pattern | High in enterocytes and glandular cells; medium in renal tubules (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); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04114-2) | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms; one chain spans residues 1–132 (UniProt) |
Section 1
Recommended LGALS2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A04114-2) appears alongside one published LGALS2 tissue microarray protocol (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 heat retrieval for the catalog antibody (datasheet: A04114-2); the published protocol used citrate pH 6.0 (PMC9242595).
Section 2
What Is the Expected LGALS2 Staining Pattern?
In paraffin-section IHC, expect predominantly cytoplasmic LGALS2 staining in intestinal enterocytes, gallbladder glandular cells and pancreatic exocrine glandular cells; renal proximal tubule cell bodies show a weaker pattern (HPA tissue IHC). HPA rates the tissue staining “Enhanced,” with medium consistency between antibody staining and RNA expression (HPA tissue IHC). LGALS2 has no annotated transmembrane segment, while UniProt does not assign a subcellular location (UniProt P05162 topology and subcellular record).
| Strong cytoplasmic staining in enterocytes or gallbladder and pancreatic glandular cells; less intense proximal tubule cell-body staining. | This matches HPA’s High calls for enterocytes in appendix, colon, duodenum, rectum and small intestine, and for gallbladder and pancreatic glandular cells. Kidney proximal tubules are Medium (HPA tissue IHC). Score the stained cell population as well as intensity (standard IHC practice). |
| Predominantly membrane-rim or nuclear chromogenic staining in an otherwise expected positive cell population. | Treat this as a compartment mismatch requiring review: HPA describes tissue IHC as cytoplasmic, and UniProt reports no transmembrane segment (HPA tissue IHC; UniProt P05162 topology). HPA also reports nucleoplasmic ICC-IF signal, so nuclear staining alone cannot establish an IHC artefact (HPA subcellular ICC-IF). |
| Prominent staining in adipocytes, adrenal glandular cells or bronchial respiratory epithelial cells. | These populations are Not detected in the supplied HPA tissue IHC record. Check for cross-reactivity or endogenous chromogen-generating activity before calling them LGALS2-positive (HPA tissue IHC; standard IHC practice). A mismatch is a reason to investigate, rather than proof of either cause. |
| Diffuse chromogen across cells and surrounding tissue, without a discernible cytoplasmic cell pattern. | The slide does not reproduce the cell-restricted cytoplasmic profile (HPA tissue IHC). Background from detection reagents or inadequate blocking can obscure a specific signal; assess a matched negative control and the no-primary control (standard IHC practice). |
| No detectable staining in an appendix, colon or gallbladder section expected to contain the listed positive cells. | The result conflicts with HPA’s High staining calls for enterocytes or gallbladder glandular cells (HPA tissue IHC). Confirm that those cells are present, then review the IHC-validated antibody and detection workflow before interpreting the sample as negative (standard IHC practice). |
💡Expected LGALS2 appearanceCall a section positive when the expected enterocytes or glandular cells show clear cytoplasmic chromogen—typically High in the listed gastrointestinal, gallbladder and pancreatic populations and Medium in proximal tubules—while staining outside those cells or as diffuse background prompts control review (HPA tissue IHC; standard IHC practice).
| Which cells and intensity should anchor scoring? | HPA calls the listed enterocytes, gallbladder glandular cells and pancreatic exocrine glandular cells High, but kidney proximal tubules Medium (HPA tissue IHC). Compare like cell populations; a weaker renal result need not contradict a stronger intestinal result. |
| What do the target annotations support? | LGALS2 is a 132-aa galectin with no annotated signal peptide, propeptide or transmembrane segment; UniProt lists one chain and no isoforms (UniProt P05162). These annotations support checking a membrane-only IHC result, but do not establish fixation sensitivity or antigen-retrieval requirements. |
| How strong is the tissue-pattern evidence? | HPA rates tissue IHC Enhanced and describes medium consistency with RNA expression; both listed antibodies have Enhanced IHC status (HPA tissue IHC; HPA antibodies). Use the observed cell pattern as a reference, while retaining the stated consistency limit. |
| Does the IF/ICC location define the paraffin-section result? | No. HPA’s ICC-IF record places LGALS2 mainly in mitochondria and additionally in nucleoplasm, whereas its tissue IHC profile is cytoplasmic (HPA subcellular ICC-IF; HPA tissue IHC). Interpret each application against its own evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive intestinal or gallbladder cells show no chromogen. | The expected cells may be absent from the section, or the IHC detection workflow may have failed (HPA tissue IHC; standard IHC practice). | Verify cell identity and section integrity, then review antibody use, antigen retrieval and detection controls according to the assay instructions (standard IHC practice). No LGALS2-specific retrieval condition is supplied. |
| Kidney proximal tubules appear weaker than intestinal enterocytes. | HPA reports Medium staining in proximal tubules and High staining in the listed enterocytes (HPA tissue IHC). | Score each population against its own expected level and inspect the cell-body pattern before changing assay conditions (HPA tissue IHC; standard IHC practice). |
| A Not detected reference population has conspicuous staining. | Cross-reactivity or endogenous detection activity is possible; the supplied HPA record does not distinguish these causes (HPA tissue IHC; standard IHC practice). | Compare a matched no-primary control and review the detection-system blocking steps; confirm the cell identity before assigning LGALS2 positivity (standard IHC practice). |
| Chromogen is diffuse across the section. | General detection background may hide the expected cell-restricted cytoplasmic pattern (HPA tissue IHC; standard IHC practice). | Inspect a no-primary control and review blocking, reagent exposure and washing under the assay protocol (standard IHC practice). Interpret intensity only after the background is resolved. |
| Membrane-rim or nuclear staining dominates the IHC slide. | That distribution differs from HPA’s cytoplasmic tissue IHC profile; ICC-IF nucleoplasmic signal is separate evidence (HPA tissue IHC; HPA subcellular ICC-IF). | Recheck the stained cell population, control slides and detection specificity before calling the compartment result positive (standard IHC practice). |
| An ICC-IF image appears mitochondrial, while a paraffin section appears cytoplasmic. | These are the respective HPA application-level descriptions, and a tissue IHC chromogen pattern does not resolve mitochondria by itself (HPA subcellular ICC-IF; HPA tissue IHC; standard IHC practice). | Record the IHC finding as cytoplasmic at the resolution of the section and assess IF/ICC using its separate guide and controls (HPA tissue IHC; HPA subcellular ICC-IF). |
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: enterocytes should stain strongly (HPA: High in colon enterocytes). Use bronchus respiratory epithelium as the negative tissue (HPA: Not detected in bronchus respiratory epithelial cells); on the colon slide, morphologically distinct non-enterocyte cells can serve as provisional internal negatives if they show only counterstain and background, without specific chromogen.
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, a concentration-matched rabbit IgG isotype control matched to the primary antibody’s clonality where known, and LGALS2-knockout tissue as a biological negative (caption: rabbit primary antibody). In colon, quench endogenous peroxidase and assess endogenous biotin if using biotin-based detection, especially in inflammatory areas (caption: biotinylated secondary and streptavidin–biotin detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A04114-2 tissue-IHC caption does not state the fixative (caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 was used for the reported paraffin-section image, but whether LGALS2 staining depends on that retrieval is unreported (caption: heat-mediated EDTA retrieval). No supplied matched evidence establishes that frozen sections or IF are easier; in colon, assess mucin and inflammatory-cell background when interpreting chromogenic staining (HPA: High in colon enterocytes).
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 type and subcellular pattern before interpreting chromogenic signal.
What retrieval should I try first if LGALS2 staining is weak?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A04114-2). The selected tissue image used that retrieval before incubation with 2 μg/mL primary antibody overnight at 4°C, so keep those conditions together for the first comparison (datasheet A04114-2). If staining remains weak, compare retrieval duration on matched sections while holding antibody concentration, detection and development constant (standard IHC practice). Include a section with strongly stained enterocytes or gallbladder glandular cells to distinguish failed retrieval from genuinely low expression in the sample (HPA: High in enterocytes and gallbladder glandular cells).
Could fixation explain variable LGALS2 staining between paraffin blocks?
The selected paraffin-section caption does not report a fixative, so LGALS2-specific sensitivity to fixation is unknown (datasheet A04114-2). Record each block’s fixation and processing history, then compare sections of similar thickness using the same EDTA pH 8.0 retrieval and staining run (datasheet A04114-2; standard IHC practice). Include a known positive tissue section in each run; inconsistent staining there points toward a processing or assay variable (HPA: High in intestinal enterocytes; standard IHC practice). If only particular blocks fail, assess tissue preservation and compare matched morphology before attributing the difference to LGALS2 biology (standard IHC practice).
How should I assess punctate or nuclear LGALS2 signal in tissue?
Assess the chromogenic signal first against the reported cytoplasmic tissue pattern in gastrointestinal epithelium, exocrine pancreas and renal tubules (HPA tissue IHC). Mitochondrial localisation with additional nucleoplasmic localisation was reported in cell-based imaging, but that finding does not establish how distinct compartments will resolve in chromogenic sections (HPA subcellular; standard IHC practice). Compare suspicious puncta or nuclear staining with a no-primary control and with adjacent cells that retain clear morphology (standard IHC practice). Record the compartment and cell type separately; do not convert diffuse brown precipitate into a mitochondrial call without independent compartment evidence (standard IHC practice).
Could an LGALS2 isoform or processing difference explain discordant staining?
The supplied record lists one 132-amino-acid chain, no isoforms, no signal peptide, no propeptide and no annotated glycosylation sites (UniProt P05162). Those annotations give no basis for assigning different tissue staining patterns to a documented LGALS2 isoform or cleavage product (UniProt P05162). The antibody epitope is not specified in the supplied caption, so its accessibility after paraffin processing cannot be predicted from the record (datasheet A04114-2). Compare EDTA pH 8.0 retrieval on matched sections and evaluate staining against expected positive cell populations before proposing an epitope-specific explanation (datasheet A04114-2; HPA tissue IHC; standard IHC practice).
How can IF help evaluate an uncertain chromogenic LGALS2 pattern?
Use IF as a separate comparison, multiplexing LGALS2 with a marker that identifies the cell population under study; in tissue sections, enterocytes and exocrine glandular cells provide reported high-expression populations (HPA tissue IHC; standard IF practice). Choose fluorophores after checking unstained tissue for autofluorescence, and include single-stain controls to assess bleed-through (standard IF practice). Because LGALS2 has no transmembrane segment, permeabilise when testing intracellular epitopes and verify access under the chosen fixation conditions (UniProt P05162 topology; standard IF practice). Compare compartment patterns cautiously: cell-based imaging reports mainly mitochondrial and additional nucleoplasmic localisation, while tissue IHC reports cytoplasmic expression (HPA subcellular; HPA tissue IHC).
What should I check when brown signal appears across the section?
Run no-primary and secondary-only controls alongside the LGALS2 section to locate signal arising from detection reagents (standard IHC practice). For the selected assay, the caption specifies a biotinylated secondary, avidin-biotin detection and DAB; check for background associated with those workflow components before changing the primary antibody (datasheet A04114-2; standard IHC practice). Apply an endogenous peroxidase block and assess whether additional blocking or washing reduces diffuse precipitate, using matched sections for each change (standard IHC practice). Preserve a positive cell population in the comparison, since excessively aggressive background reduction can also conceal expected enterocyte or glandular staining (HPA: High in enterocytes and gallbladder glandular cells; standard IHC practice).
How should I quantify LGALS2 staining across different tissues? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored compartment and cell population before measuring LGALS2, using the reported cytoplasmic tissue pattern as the initial annotation guide (HPA tissue IHC). For epithelial samples, report the percentage of positive target cells and an H-score from 0–300, recording intensity thresholds and reader agreement (standard IHC practice). Normalise counts to the number of evaluable target cells, or report positive-cell density per mm² of annotated viable tissue when cell counts are impractical (standard IHC practice). Keep regions with necrosis, folds or section edges out of the denominator, and compare batches only after checking a shared control section (standard IHC practice).
When is an unexpected LGALS2-positive region convincing?
A convincing result follows intact cells and a reproducible compartment pattern, with appropriate signal in reported positive populations such as intestinal enterocytes or pancreatic exocrine glandular cells (HPA tissue IHC; standard IHC practice). Treat isolated nuclear staining cautiously because tissue IHC reports cytoplasmic expression, although cell-based imaging also reports nucleoplasmic localisation (HPA tissue IHC; HPA subcellular). Inspect unexpected cell types against no-primary controls and adjacent morphology, particularly around section edges or necrotic areas where staining artefacts can concentrate (standard IHC practice). If DAB persists without primary antibody, investigate endogenous enzyme activity or detection background before assigning LGALS2 positivity (standard IHC practice).
Boster reagents
Best LGALS2 / Galectin-2 IHC Antibodies
Two anti-LGALS2 antibodies have IHC images from human paraffin sections; both list IF/ICC, and one lists human, mouse and rat reactivity (catalog applications, reactivity and 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 reactivity and has IHC images from paraffin sections of human pancreatic and rectal cancer, plus IF data from A549 cells (catalog reactivity and image captions). A04114 lists human, mouse and rat reactivity and has an IHC image from a paraffin section of human heart, plus IF data from COS7 cells (catalog reactivity and image captions).
Which to pick: For tissue IHC, A04114-2 has documented retrieval and detection conditions for human paraffin sections: EDTA pH 8.0 retrieval, 2 μg/ml primary antibody and DAB detection (A04114-2 IHC image captions). For IF/ICC, both SKUs list the applications; A04114-2 documents 5 μg/ml in A549 cells, while A04114 shows COS7 IF with peptide blocking (catalog applications and IF image captions). Choose A04114 when listed mouse or rat reactivity is needed; its IHC caption documents human heart only, and neither SKU’s IHC caption reports the tissue fixative (catalog reactivity and IHC image captions).
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 in GI, gallbladder, pancreatic and renal cells (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); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04114-2); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one chain spans residues 1–132 (UniProt); Regulation: No expression regulator annotated (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.