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GLO1 / Lactoylglutathione lyase · IHC design guide
Design Immunohistochemistry for GLO1
Plan chromogenic IHC for GLO1 in paraffin sections using the catalog antibody’s validated conditions. Compare cytoplasmic staining in Leydig and glandular cells with appropriate negative controls, and score the relevant cell types separately (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GLO1 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 several tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic; strong in Leydig and selected glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A01703-1) | |
| Positive control | Epididymis+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 | High staining is confined to specific cell types (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unspecified (UniProt) |
Section 1
Recommended GLO1 IHC & IF Protocols
The catalog antibody has a citrate pH 6 IHC-P protocol (datasheet A01703-1). The published options below cover prostate, aortic sinus, and liver or heart sections (PMC8304603; PMC4693023; PMC5813374).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A01703-1) |
| Fixation | Image fixative and duration unreported (datasheet A01703-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A01703-1) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01703-1) |
| Primary antibody | Rabbit anti-GLO1, 0.5-1μg/ml (datasheet A01703-1) |
| Primary incubation | Overnight at 4 °C (datasheet A01703-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01703-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GLO1-positive staining in glandular cells of epididymis (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues, highly abundant in Leydig cells and in in prostate, epididymis and thyroid glandular cells. No signal in the no-primary control. |
💡Decision noteStart with heat retrieval at pH 6 (datasheet A01703-1); the aortic sinus and liver protocols also use pH 6 retrieval (PMC4693023; PMC5813374).
Section 2
What Is the Expected GLO1 Staining Pattern?
For paraffin-section IHC, expect predominantly cytoplasmic GLO1 staining, strongest in epididymal, prostate and thyroid glandular cells and testicular Leydig cells (HPA tissue IHC: High). HPA rates the tissue profile Enhanced, with medium antibody–RNA consistency and agreement between paired antibodies (HPA tissue IHC: Enhanced). GLO1 has no transmembrane segment; UniProt does not assign it a subcellular location (UniProt Q04760: topology and subcellular annotation).
| Clear cytoplasmic stain in glandular cells of epididymis, prostate or thyroid, or in testicular Leydig cells (HPA tissue IHC: High). | This matches the reported high-signal cell types and cytoplasmic tissue profile (HPA tissue IHC). Judge the named cells against neighboring cells on the same section; staining across an entire tissue is less informative than its cellular distribution (general IHC practice). |
| Predominantly nuclear or sharply membrane-only stain replaces the expected cytoplasmic IHC pattern (HPA tissue IHC: cytoplasmic). | Treat an isolated compartment shift as a reason to check specificity and detection conditions, rather than calling it GLO1 from location alone (general IHC practice). Nucleoplasm and plasma membrane are supported in ICC-IF, so those locations cannot be declared intrinsically impossible (HPA subcellular ICC-IF). |
| Strong stain appears in adipocytes, cardiomyocytes or skeletal myocytes while the expected positive cells are weak (HPA tissue IHC: Not detected in those cell types). | This conflicts with the reported cell-level IHC pattern and raises concern for cross-reactivity or endogenous detection activity (HPA tissue IHC; general IHC practice). A Not detected call applies to the specified cells under HPA's assay conditions, not to every cell in that tissue. |
| Chromogen coats extracellular areas or many unrelated cell types without a readable intracellular pattern (general IHC practice). | Diffuse background limits interpretation even if a positive tissue is present. Assess a no-primary control and the detection reagents, blocking and washes before assigning that color to GLO1 (general IHC practice); HPA's cell-level profile cannot validate featureless staining. |
| No stain is visible in glandular cells of epididymis, prostate or thyroid, or in testicular Leydig cells (HPA tissue IHC: High). | A blank expected-positive cell population makes a negative call elsewhere unreliable. Check tissue integrity and an on-run positive control, then review the antibody and IHC detection workflow (general IHC practice). HPA's High rating is an expectation, not a guarantee for every section. |
💡Expected GLO1 appearanceCall a convincing IHC positive when the expected glandular cells or Leydig cells show prominent cytoplasmic color (HPA tissue IHC: High, cytoplasmic); widespread cell-independent haze or dominant staining of HPA Not detected cell types is suspect (HPA tissue IHC; general IHC practice).
| Cell type and tissue selection (HPA tissue IHC). | Epididymal, prostate and thyroid glandular cells and testicular Leydig cells are High; adrenal and appendix glandular cells, marrow hematopoietic cells and bronchial basal cells are Medium (HPA tissue IHC). Score the specified cells, since adjacent cell populations need not share a rating. |
| Negative comparison cells (HPA tissue IHC). | Adipocytes, cardiomyocytes, skeletal myocytes and several other listed cell types are Not detected (HPA tissue IHC). Use those calls as cell-specific comparisons; they do not establish a whole-organ negative control or prove that every unstained cell lacks GLO1. |
| IHC evidence and antibody choice (HPA tissue IHC; HPA antibodies). | The tissue profile is Enhanced, with medium antibody–RNA consistency and supporting agreement between paired antibodies (HPA tissue IHC). Two listed mouse monoclonals have Enhanced IHC validation; the listed rabbit polyclonal has no IHC status supplied (HPA antibodies). |
| Compartment evidence differs by application (HPA tissue IHC; HPA subcellular ICC-IF). | Tissue IHC is described as cytoplasmic, while ICC-IF supports nucleoplasm, plasma membrane and cytosol (HPA). IF localization informs a compartment discrepancy but does not, by itself, validate a nuclear or membrane-only pattern in paraffin-section chromogenic IHC. |
| Protein features and epitope uncertainty (UniProt Q04760). | GLO1 has two isoforms, a VOC domain at residues 31–177, no signal peptide or transmembrane segment, and no annotated subcellular location (UniProt Q04760). Without the antibody epitope and isoform coverage, these features cannot predict retrieval conditions or an isoform-specific staining pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected-positive cells are blank (HPA tissue IHC: High cell types). | A failed run, unsuitable antibody conditions or poor tissue preservation can prevent an interpretable IHC result (general IHC practice); the supplied sources give no GLO1-specific fixation sensitivity. | Check an on-run positive section, primary-antibody use, detection reagents and tissue morphology. Review the catalog antibody's IHC-P instructions for retrieval and dilution before adjusting conditions (general IHC practice). |
| Broad, weak color obscures cell boundaries (general IHC practice). | Nonspecific primary binding, detection background or inadequate washing are possible general IHC causes; the HPA pattern alone cannot identify which applies. | Compare a no-primary control, inspect blocking and wash steps, and adjust antibody or detection conditions according to the IHC-P instructions (general IHC practice). Interpret GLO1 only when cellular staining separates from background. |
| Chromogen persists in the no-primary control (general IHC practice). | The primary antibody cannot account for signal in this control. Endogenous detection activity or detection-system background is possible (general chromogenic IHC practice). | Inspect the detection system and its appropriate endogenous-activity blocking step, then repeat the control (general IHC practice). Do not score persistent control color as GLO1. |
| A nuclear or membrane-only pattern dominates paraffin-section IHC (HPA tissue IHC: cytoplasmic). | The pattern differs from HPA's tissue-IHC summary, although those compartments are supported in ICC-IF (HPA subcellular ICC-IF). The discrepancy alone does not identify its cause. | Recheck the expected-positive cell types, no-primary control and IHC antibody validation. Record the compartment discrepancy rather than treating IF localization as confirmation of the IHC result (HPA tissue IHC; HPA antibodies; general IHC practice). |
| Cells rated Not detected stain strongly (HPA tissue IHC). | Cross-reactivity or detection background is possible (general IHC practice). HPA's Not detected calls concern named cell populations and do not certify a completely blank tissue. | Identify the stained cell type on the counterstained section, compare a no-primary control, and check staining in a High reference cell population from the same run (HPA tissue IHC; general IHC practice). |
| Signal varies within an expected-positive glandular population (HPA tissue IHC: High). | Section quality or uneven staining may complicate comparison (general IHC practice); HPA's High category does not require identical intensity in every cell. | Compare intact areas and another section processed in the same run. Score the named cells and cytoplasmic pattern before interpreting variation as biological (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing GLO1 staining
A01703-1 · IHCIHC analysis of GLO1 using anti-GLO1 antibody (A01703-1). GLO1 was detected in paraffin-embedded section of human lung cancer tissue. Heat m
A01703-1 · IHCIHC analysis of GLO1 using anti-GLO1 antibody (A01703-1). GLO1 was detected in paraffin-embedded section of rat small intestine tissue. Heat
A01703-1 · IFIF analysis of GLO1 using anti-GLO1 antibody (A01703-1) and anti-Tubulin Alpha antibody (M03989-3). GLO1 was detected in immunocytochemical
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000124767-GLO1/tissue/cerebral+cortex#imid_10264477
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000124767-GLO1/tissue/colon#imid_10264403
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000124767-GLO1/tissue/liver#imid_10264396
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000124767-GLO1/tissue/kidney#imid_10264413
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB040541; green = GLO1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB040541; green = GLO1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GLO1 IHC & IF
🧪Run prostate first and expect glandular cells to stain (HPA: High in prostate glandular cells). Use esophageal squamous epithelium as the negative tissue (HPA: Not detected in esophageal squamous epithelial cells); on the prostate slide, use stromal cells as an internal background comparator, without assuming they are GLO1-negative because their staining status is not supplied (HPA: prostate glandular cells only).
Positive control tissue: Epididymis (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GLO1 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (supported), Plasma membrane (supported), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (selected-SKU caption: rabbit primary antibody). Confirm specificity with a GLO1 knockout specimen where available, and check endogenous peroxidase and biotin signal because the reported IHC detection uses SABC and DAB (selected-SKU caption: SABC/DAB).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption does not state a fixative (selected-SKU caption: fixative unreported). The reported IHC procedure uses heat retrieval in citrate buffer at pH 6 for 20 minutes, but retrieval dependence has not been established (selected-SKU caption: retrieval conditions). The supplied evidence does not establish whether frozen sections or IF are easier; score prostate glandular cells separately from stromal background and check detection-only controls for peroxidase or biotin signal (HPA: High in prostate glandular cells; selected-SKU caption: SABC/DAB).
HPA tissue IHC evidence for GLO1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Paired antibodies with high similarity supports the protein expression profile.). 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 |
|---|---|---|---|---|
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Prostate | Glandular cells | High | Protein (IHC) | HPA → |
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
| Thyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GLO1 IHC Tips
Troubleshoot GLO1 staining in paraffin sections by checking retrieval, cell type, compartment and detection controls before comparing staining intensity.
Which retrieval conditions should I try first for weak GLO1 staining?
Start with heat-mediated retrieval in citrate buffer at pH 6 (datasheet A01703-1). The catalog antibody’s paraffin-section image used 20 minutes of retrieval under those conditions, followed by 1 µg/mL primary antibody overnight at 4°C (caption A01703-1). Check that sections remained covered by buffer throughout heating and cooled consistently before staining (standard IHC practice). If staining remains weak, compare a small retrieval time series while keeping antibody concentration and detection conditions fixed (standard IHC practice). Interpret gains alongside tissue morphology and a matched negative control, since stronger staining alone does not establish specificity (standard IHC practice).
Could fixation explain inconsistent GLO1 staining across paraffin blocks?
Target-specific sensitivity of GLO1 to fixation is unknown from the supplied evidence: the catalog tissue-IHC caption does not state a fixative (caption A01703-1). Record each block’s fixative and fixation duration before comparing staining between blocks (standard IHC practice). For a controlled comparison, section blocks at the same thickness and hold citrate retrieval at pH 6 for 20 minutes, matching the documented image conditions (caption A01703-1). Include a consistently processed reference section in each run to detect shifts in staining (standard IHC practice). Do not assign a fixation effect to GLO1 from staining intensity alone; processing and detection also influence the result (standard IHC practice).
How should I assess GLO1 staining outside the cytoplasm?
Expect cytoplasmic staining in several tissues by IHC, with high abundance reported in Leydig cells and selected glandular cells (HPA tissue IHC). Nucleoplasm, plasma membrane and cytosol are supported locations in cell-based imaging, while UniProt does not annotate a subcellular location for GLO1 (HPA subcellular; UniProt Q04760). For paraffin sections, first score cytoplasmic signal against cellular morphology and a negative control (HPA tissue IHC; standard IHC practice). Review convincing nuclear or membrane staining separately rather than folding it into the cytoplasmic score (standard IHC practice). GLO1 has no annotated transmembrane segment, so a sharp membrane outline warrants additional specificity checks (UniProt Q04760 topology; standard IHC practice).
Can epitope differences explain variable GLO1 staining?
GLO1 has 2 annotated isoforms, and the supplied evidence does not map the catalog antibody’s epitope to either one (UniProt Q04760; supplied catalog evidence). Its VOC domain spans residues 31–177, and annotated modified residues include phosphothreonine 107 and alternate modifications at 139 and 148 (UniProt Q04760). These annotations identify features to check against an epitope map; they do not establish which feature affects staining by this antibody (UniProt Q04760; supplied catalog evidence). If an epitope map becomes available, compare its sequence with both isoforms before interpreting a negative IHC result (standard IHC practice). Validate unexpected patterns with an independent antibody or another specificity control (standard IHC practice).
How can IF help check cell type and compartment assignments?
Use the separate IF/ICC guide for experimental conditions; the evidence here supports GLO1 in the nucleoplasm, plasma membrane and cytosol in cell-based imaging (HPA subcellular). In multiplex IF, pair GLO1 with a marker that identifies the expected cell type in the specimen, such as a Leydig-cell or glandular-cell marker where applicable (HPA tissue IHC; standard IF practice). Choose fluorophores after measuring tissue autofluorescence, and include single-channel controls to assess spectral bleed-through (standard IF practice). Plan permeabilisation around epitope access: an intracellular epitope needs membrane access, whereas a surface-facing epitope may be assessed before permeabilisation (standard IF practice). The catalog antibody’s tissue-IHC caption does not establish an IF/ICC fixation or permeabilisation condition (caption A01703-1).
What should I check when GLO1 DAB staining is diffuse?
Compare the stained section with a no-primary control and inspect whether diffuse colour follows tissue boundaries, damaged areas or the expected cells (standard IHC practice). The catalog image used 10% goat serum block, biotinylated secondary antibody, a streptavidin-biotin complex and DAB detection (caption A01703-1). For that detection format, check endogenous peroxidase blocking and consider whether endogenous biotin contributes signal; these are general workflow concerns, not demonstrated GLO1-specific effects (standard IHC practice). Adjust blocking, washes and primary concentration one variable at a time while retaining a reference section (standard IHC practice). Compare the outcome with the reported cytoplasmic tissue pattern before accepting stronger DAB colour as GLO1 signal (HPA tissue IHC).
How should I compare GLO1 IHC staining between specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and scoring compartment before measuring colour: HPA reports cytoplasmic tissue staining, including high signal in Leydig cells and selected glandular cells (HPA tissue IHC). Within that population, record the percentage of positive cells and intensity, or calculate an H-score using the same intensity bins for every specimen (standard IHC practice). If counting discrete positive cells, report density per mm² of viable, evaluable tissue and document excluded areas (standard IHC practice). Normalise comparisons to the relevant cell population or tissue area, rather than the whole section when cellular composition differs (standard IHC practice). Keep staining batches, imaging and thresholds consistent and include a reference section across runs (standard IHC practice).
How can I distinguish true GLO1 signal from staining artefact?
Start with cell identity and compartment: tissue IHC reports cytoplasmic GLO1, with high staining in Leydig cells and glandular cells of the prostate, epididymis and thyroid (HPA tissue IHC). Signal confined to section edges, necrotic regions or a compartment that conflicts with the expected tissue pattern needs closer review (HPA tissue IHC; standard IHC practice). Check a no-primary control for DAB colour from endogenous enzyme activity or the detection system (standard IHC practice). Compare suspect cells with nearby morphology and an independently stained reference section under the same imaging conditions (standard IHC practice). Treat a plausible pattern as supportive evidence, then confirm unexpected distributions with an independent specificity control (standard IHC practice).
Boster reagents
Best GLO1 / Lactoylglutathione lyase IHC Antibodies
Anti-GLO1 antibody A01703-1 has paraffin-section IHC images from human and rat tissues (catalog IHC captions) and an IF/ICC image from A549 cells (catalog IF caption).
Real IHC data
Anti-GLO1 Antibody ®
Cat # A01703-1
A01703-1 will render with an IHC figure from a human lung cancer paraffin section; its other IHC captions show human mammary cancer, rat small intestine, and rat spleen paraffin sections (catalog IHC captions). Its IF/ICC caption shows staining in A549 cells (catalog IF caption).
Which to pick: Choose A01703-1 for paraffin-section IHC or A549 IF/ICC because both applications have image data for this SKU (catalog IHC and IF captions). It is polyclonal and lists Human, Mouse, and Rat reactivity, though the supplied images show human and rat IHC and A549 IF/ICC only (catalog dilution listing; catalog IHC and IF captions). The IHC captions report paraffin sections but do not report the fixative (catalog IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q04760 (LGUL_HUMAN, Lactoylglutathione lyase).
- Human Protein Atlas. GLO1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. GLO1 subcellular location (ICC-IF): Localized to the nucleoplasm, plasma membrane and cytosol..
- Human Protein Atlas. GLO1 antibody validation summary (3 antibodies).
- Glyoxalase 1 Expression as a Novel Diagnostic Marker of High-Grade Prostatic Intraepithelial Neoplasia in Prostate Cancer. Cancers 2021 — PMC8304603.
- A Glyoxalase-1 Knockdown Does Not Have Major Short Term Effects on Energy Expenditure and Atherosclerosis in Mice. Journal of diabetes research 2016 — PMC4693023.
- Proteomic identification and characterization of hepatic glyoxalase 1 dysregulation in non-alcoholic fatty liver disease. Proteome science 2018 — PMC5813374.
- Characterisation of glyoxalase I in a streptozocin-induced mouse model of diabetes with painful and insensate neuropathy. Diabetologia 2011 — PMC3762253.
- PubMed PMID:7684374 — UniProt-cited evidence.
- PubMed PMID:8449929 — UniProt-cited evidence.
- PubMed PMID:8670058 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GLO1 (Lactoylglutathione lyase). Expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A01703-1). Validated IHC antibody: A01703-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Epididymis (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A01703-1); 2. Fixation: Image fixative and duration unreported (datasheet A01703-1); 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: Citrate pH 6 , 20 min (datasheet A01703-1); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01703-1); 8. Primary antibody: Rabbit anti-GLO1, 0.5-1μg/ml (datasheet A01703-1); 9. Primary incubation: Overnight at 4 °C (datasheet A01703-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: High staining is confined to specific cell types (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences unspecified (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q04760, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm, plasma membrane and cytosol., Datasheet A01703-1, PMC8304603, PMC4693023. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GLO1/immunohistochemistry-ihc-guide.