This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
GLO1 / Lactoylglutathione lyase · IHC design guide
Design Immunohistochemistry for GLO1
Plan chromogenic GLO1 IHC in paraffin sections using high staining in Leydig cells and prostate, epididymal or thyroid glandular cells as reference patterns (HPA tissue IHC). Compare cell-specific cytoplasmic staining with appropriate controls and keep fixation consistent across samples (HPA tissue IHC; standard IHC practice).
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 signal in Leydig and 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 tissue fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining has medium RNA consistency (HPA tissue IHC) | |
| Regulation | Expression regulation is unannotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended GLO1 IHC & IF Protocols
The catalog antibody protocol is followed by three published chromogenic IHC protocols for GLO1 in paraffin 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-mediated citrate retrieval at pH 6 (datasheet A01703-1); the published aortic sinus and liver protocols also use pH 6 retrieval (PMC4693023 methods; PMC5813374 methods).
Section 2
What Is the Expected GLO1 Staining Pattern?
Expect predominantly cytoplasmic GLO1 staining in glandular cells of epididymis, prostate and thyroid gland, and in testicular Leydig cells (HPA tissue IHC: High; reliability Enhanced). HPA also observes medium staining in selected other cell populations (HPA tissue IHC). GLO1 has no annotated transmembrane segment, while UniProt does not assign it a subcellular location (UniProt Q04760 topology and subcellular record).
| Strong cytoplasmic staining in epididymal, prostate or thyroid glandular cells, or testicular Leydig cells. | This matches the High cell-specific IHC pattern (HPA tissue IHC). Judge the named cells, since an entire tissue section contains other cell types. HPA rates the tissue profile Enhanced, with medium agreement between antibody staining and RNA data (HPA tissue IHC reliability). |
| Predominantly extracellular or luminal deposit, with little staining in the expected cells. | That distribution does not match HPA's cytoplasmic tissue pattern (HPA tissue IHC). Inspect morphology and the detection controls before interpreting it as GLO1. Nuclear or membrane signal alone is less decisive: those locations are supported by ICC-IF, but their prominence in paraffin IHC is not established here (HPA subcellular ICC-IF). |
| Strong staining in adipocytes, cardiomyocytes or skeletal myocytes. | These cell types are listed as Not detected by HPA tissue IHC (HPA tissue IHC). Unexpected chromogen may reflect antibody cross-reactivity or endogenous detection activity (general IHC practice); the image alone cannot distinguish them. Recheck cell identity and compare appropriate detection controls. |
| Diffuse colour across cells and surrounding tissue, obscuring cell boundaries. | A uniform haze is difficult to reconcile with the named high-staining cell populations (HPA tissue IHC). Background from detection chemistry or nonspecific antibody binding is possible (general IHC practice). Interpret a positive result only where cellular morphology and compartment can still be resolved. |
| No staining in an otherwise interpretable epididymis, prostate, testis or thyroid section. | A missing signal conflicts with the listed High populations if those cells are present (HPA tissue IHC). Confirm their presence on the section, then assess the staining run and antibody conditions (general IHC practice). One blank field cannot establish absent GLO1 expression throughout a tissue. |
💡Expected GLO1 appearanceCall a paraffin IHC result positive when cytoplasmic colour is clear in the expected glandular cells or Leydig cells, with High staining in those populations (HPA tissue IHC); widespread extracellular deposit or strong colour in HPA-listed undetected cells warrants control review before a GLO1 call (HPA tissue IHC; general IHC practice).
| Cell population and reference intensity | HPA lists glandular cells in epididymis, prostate and thyroid, plus testicular Leydig cells, as High; adrenal and appendix glandular cells, marrow hematopoietic cells and bronchial basal cells are Medium (HPA tissue IHC). Use the named cell population when comparing sections; a medium population need not resemble a high one. |
| Evidence behind the IHC pattern | The tissue profile is rated Enhanced, with medium consistency between staining and RNA expression; similar staining from paired antibodies supports the profile (HPA tissue IHC reliability). CAB040541 and CAB040542 each have Enhanced IHC validation (HPA antibodies). This supports the observed pattern without proving every individual positive cell is specific. |
| Isoforms and antibody epitope | UniProt records 2 GLO1 isoforms and a VOC domain spanning residues 31–177 (UniProt Q04760). The payload gives no epitope location or isoform coverage for an IHC antibody, so a difference between antibodies cannot be assigned to isoform recognition from these records. Check the antibody's documented epitope before making that interpretation. |
| IF/ICC Q&A: where may GLO1 appear? | HPA supports nucleoplasm, plasma membrane and cytosol in ICC-IF images from A-431, U-251MG and U2OS (HPA subcellular ICC-IF). That answers the IF localisation question; it does not set a separate paraffin IHC scoring rule. UniProt lists no subcellular location and no transmembrane segment (UniProt Q04760). |
| Retrieval and detection conditions | No GLO1-specific retrieval setting or fixation sensitivity is supplied by UniProt or HPA. For chromogenic paraffin IHC, compare candidate retrieval conditions using the same known-positive and low-signal reference cells, with detection controls (general IHC practice; HPA tissue IHC for reference cells). Do not infer a target-specific fixation effect from a staining level. |
| Situation | Likely cause | Next action |
|---|---|---|
| High-reference cells show no colour. | The intended cells may be absent from the viewed field, or the staining run may have failed (general IHC practice); HPA lists these populations as High (HPA tissue IHC). | Locate the named glandular cells or Leydig cells on the section first. Review the run's positive control, antibody application and detection steps before calling the sample negative (general IHC practice). |
| Signal is faint in a High reference but visible in Medium populations. | The observed ranking disagrees with the reference profile, which assigns different levels to specific cell populations (HPA tissue IHC). Section composition or staining conditions may affect the comparison (general IHC practice). | Compare equivalent, intact cell populations on the same run. Document intensity by cell type and review antibody and detection conditions before revising the biological interpretation (general IHC practice). |
| Adipocytes or muscle cells stain strongly. | HPA records adipocytes, cardiomyocytes and skeletal myocytes as Not detected (HPA tissue IHC). Cross-reactivity or endogenous detection activity could produce an apparent positive (general IHC practice). | Confirm the stained cell type and inspect a detection control without primary antibody. If colour persists, investigate the detection system; if it does not, reassess antibody specificity (general IHC practice). |
| Background colour obscures boundaries. | Nonspecific antibody binding or detection background may be masking the cellular distribution (general IHC practice). The reference pattern is cytoplasmic in selected tissue cells (HPA tissue IHC). | Check the no-primary detection control, blocking and antibody concentration; compare morphology in counterstained sections. Score GLO1 only where the expected cells and their cytoplasm remain distinguishable (general IHC practice; HPA tissue IHC). |
| Colour pools in lumina or outside cells. | That distribution differs from the cytoplasmic tissue profile (HPA tissue IHC). Deposited detection product or section artefact is possible (general IHC practice); the supplied records do not establish secreted GLO1 staining. | Review adjacent cellular morphology and detection controls. Require a reproducible signal in the HPA-listed positive cells before attributing extracellular colour to GLO1 (HPA tissue IHC; general IHC practice). |
| A prominent nuclear or membrane signal dominates the IHC slide. | HPA supports both locations in ICC-IF, alongside cytosol, but describes the tissue IHC profile as cytoplasmic (HPA subcellular ICC-IF; HPA tissue IHC). The two applications do not establish identical compartment intensities. | Record the compartment and cell type separately. Compare cytoplasmic staining in a listed High population and review IHC detection controls before treating a nuclear or membrane dominant pattern as the expected paraffin result (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 epididymis first; its glandular cells should stain for GLO1 (HPA: High in epididymis glandular cells). Use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipocytes); on the epididymis slide, assess any non-glandular cells for absent staining, but HPA does not designate an internal negative cell population (HPA: epididymis glandular-cell row).
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 slide and a rabbit IgG isotype control matched to the primary antibody’s clonality if known (selected A01703-1 caption: rabbit primary); confirm specificity with GLO1 knockout material or peptide competition if a suitable peptide is available (standard IHC practice). Block endogenous peroxidase and check endogenous biotin background when using the caption’s SABC/DAB detection in epididymis (selected A01703-1 caption: SABC/DAB).
⚠️Feasibility: The selected A01703-1 paraffin-section caption reports citrate retrieval at pH 6 for 20 minutes, but does not report the fixative or a target-specific fixation window or effect; retrieval dependence beyond that example is unreported (selected A01703-1 caption). HPA has ICC-IF images for GLO1, but the supplied evidence does not establish whether frozen sections or IF are easier than paraffin IHC (HPA: GLO1 subcellular ICC-IF record). No epididymis-specific artefact is documented in the supplied tissue evidence (HPA: epididymis glandular-cell row).
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 section chromogenic IHC using the catalog antibody’s tissue result and independent expression data.
What retrieval should I try first if GLO1 staining is weak in paraffin sections?
Start with heat mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A01703-1). This is the condition used for the catalog antibody’s staining of a paraffin section of human lung cancer tissue (datasheet A01703-1). Keep section thickness, heating and cooling consistent while comparing a weak sample with a positive control (standard IHC practice). If staining remains weak, test an alternative retrieval condition on adjacent sections and compare signal with tissue damage and background (standard IHC practice). Record any change as a local optimisation, since the supplied tissue result supports only the citrate condition (datasheet A01703-1).
Could fixation explain weak or uneven GLO1 staining?
The catalog image identifies a paraffin section but does not report its fixative, so GLO1 sensitivity to fixation is unknown (datasheet A01703-1). Compare sections with documented, consistent fixation histories before attributing unequal staining to the antibody or biology (standard IHC practice). Use the reported citrate pH 6, 20 minute retrieval condition as a consistent starting point across those sections (datasheet A01703-1). Examine morphology alongside staining: uneven preservation can make intensity comparisons unreliable even when the detection steps match (standard IHC practice). Neither tissue expression patterns nor GLO1’s sequence features establish a target specific fixation effect here (HPA tissue IHC; UniProt Q04760).
Is nuclear or membrane staining plausible when I expect cytoplasmic GLO1?
Cytoplasmic staining across several tissues is the reported tissue IHC pattern (HPA tissue IHC). Independent cell imaging also supports nucleoplasm, plasma membrane and cytosol localisation, so nuclear or membrane signal deserves assessment rather than automatic rejection (HPA subcellular). GLO1 has no annotated transmembrane segment; membrane staining alone therefore does not establish that the protein spans the membrane (UniProt Q04760 topology). Compare compartments within clearly identified cells using the same counterstain and exposure conditions, then check whether the pattern persists in replicate sections (standard IHC practice). Judge unusual staining against cell specific controls, because tissue level localisation and cell imaging provide different kinds of evidence (HPA tissue IHC; HPA subcellular).
Can an epitope or GLO1 isoform difference explain discordant IHC staining?
GLO1 has 2 annotated isoforms, but the supplied catalog image does not map this antibody’s epitope or establish isoform selectivity (UniProt Q04760; datasheet A01703-1). The protein contains a VOC domain at residues 31–177 and reported modified residues, including positions 88 and 107 (UniProt Q04760). An epitope overlapping a variable or modified region could affect recognition, but that remains a hypothesis until its sequence is known (UniProt Q04760; standard IHC practice). Request epitope mapping or compare independently validated antibodies before assigning a staining difference to an isoform (standard IHC practice). Keep retrieval and detection matched during that comparison so technical changes do not confound it (standard IHC practice).
How should I check a disputed GLO1 pattern by multiplex IF?
For a separate IF/ICC check, pair GLO1 with a validated marker for the cell type being examined; glandular cells and Leydig cells provide documented positive contexts (HPA tissue IHC). Choose spectrally separated fluorophores and assess tissue autofluorescence in an unstained control before interpreting apparent colocalisation (standard IF practice). HPA cell imaging supports nucleoplasm, plasma membrane and cytosol localisation, while GLO1 has no annotated transmembrane segment (HPA subcellular; UniProt Q04760 topology). Permeabilise when testing an intracellular epitope; for a proposed surface epitope, establish its accessibility before detergent treatment, since this antibody’s epitope orientation is unreported (standard IF practice; datasheet A01703-1). Confirm multiplex signals with single stain controls and cell matched comparisons (standard IF practice).
How can I distinguish GLO1 signal from diffuse DAB background?
The catalog tissue result used 10% goat serum block, 1 µg/mL primary antibody overnight at 4°C, biotinylated secondary and DAB development (datasheet A01703-1). Reproduce those documented conditions first, then compare a no primary control and a known positive section through the same detection sequence (standard IHC practice). Include a peroxidase block and inspect endogenous pigment or enzyme related colour before calling weak diffuse DAB signal positive (standard chromogenic IHC practice). If background persists, adjust blocking, washing or primary concentration one variable at a time, preserving a paired control (standard IHC practice). Evaluate signal within identified cells; HPA reports uneven cell type expression across tissues (HPA tissue IHC).
What scoring method best captures heterogeneous GLO1 IHC staining? ⚠ ANSWER MARKED FOR VERIFICATION
Score GLO1 within predefined cell populations because HPA reports high glandular cell staining in prostate, epididymis and thyroid gland, and high Leydig cell staining in testis (HPA tissue IHC). For intensity, an H-score combines the percentage of cells at each 0–3 intensity level into a 0–300 score (standard IHC practice). Alternatively report percentage positive cells or positive cell density per mm² when intensity or cell abundance is the primary question (standard IHC practice). Normalise counts to viable, evaluable cells or tissue area, and exclude folds, edges and necrosis by a prespecified rule (standard IHC practice). Keep retrieval, DAB development and scoring thresholds consistent across compared sections (standard IHC practice).
When is an unexpected GLO1 positive area likely to be artefact?
First verify that staining lies in intact, identifiable cells rather than section edges, folds or necrotic areas (standard IHC practice). Cytoplasmic tissue staining is documented, while cell imaging also supports nucleoplasm and plasma membrane, so compartment alone cannot classify every unexpected signal (HPA tissue IHC; HPA subcellular). Compare the questioned cell population with a positive control such as prostate glandular cells and a reported undetected population such as adipocytes (HPA tissue IHC). Test whether apparent DAB signal survives an appropriate peroxidase block and disappears in the no primary control (standard chromogenic IHC practice). Treat persistent discordance as unresolved until cell identity, controls and section quality agree (standard IHC practice).
Boster reagents
Best GLO1 / Lactoylglutathione lyase IHC Antibodies
The IHC-validated GLO1 antibody has paraffin-section images from human and rat tissues (catalog IHC captions) and an IF/ICC image from human cells (catalog IF caption).
Real IHC data
Anti-GLO1 Antibody ®
Cat # A01703-1
A01703-1 has IHC images from human lung cancer and mammary cancer tissues and rat small intestine and spleen, all in paraffin sections (catalog IHC captions). It also has an IF/ICC image from A549 cells (catalog IF caption); the listed reactivity is human, mouse and rat (catalog reactivity).
Which to pick: For tissue IHC, choose A01703-1: its images show staining in human and rat paraffin sections (catalog IHC captions); the fixative is unreported (catalog IHC captions). For IF/ICC, A01703-1 has an A549 cell image (catalog IF caption) and is listed for IF and ICC (catalog applications). For cross-species studies, A01703-1 is a polyclonal antibody (catalog dilution data) listed as reactive with human, mouse and rat (catalog reactivity), though the supplied IHC images cover human and rat only (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 tissue fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Antibody staining has medium RNA consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unknown (UniProt); Regulation: Expression regulation is unannotated (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.