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GLRX / Glutaredoxin-1 · IHC design guide
Design Immunohistochemistry for GLRX
Plan chromogenic IHC for GLRX using kidney tubules as a high-staining reference (HPA tissue IHC). Compare cytoplasmic staining across sections (HPA tissue IHC), starting the IHC-validated antibody at 2–5 μg/ml (datasheet A03392-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GLRX 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 in tissue sections (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in several tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03392-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Breast+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Regulation not annotated (UniProt) | |
| Isoform / epitope | No isoforms annotated; chain spans residues 2–106 (UniProt) |
Section 1
Recommended GLRX IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by 3 published GLRX chromogenic IHC protocols (datasheet: A03392-1; PMC8805200; PMC10916419; PMC8442864).
| Sample | Paraffin-embedded human kidney tissue; fixative not specified (datasheet A03392-1) |
| Fixation | Image fixative and duration unreported (datasheet A03392-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A03392-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03392-1) |
| Primary antibody | Rabbit anti-GLRX, 2-5 μg/ml (datasheet A03392-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03392-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03392-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GLRX-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues, including kidney, gastrointestinal tract and placenta. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 for the catalog antibody (datasheet: A03392-1). The published protocols used citrate or citric acid retrieval (PMC8805200; PMC10916419; PMC8442864).
Section 2
What Is the Expected GLRX Staining Pattern?
GLRX should appear mainly in the cytoplasm of stained cells (UniProt P35754: cytoplasm; HPA: cytoplasmic tissue expression). High IHC staining is reported in kidney tubule cells and several glandular or epithelial cell populations (HPA: tissue IHC). Treat these as useful comparators, not universal thresholds: HPA rates the tissue staining Approved but reports medium consistency with RNA expression (HPA: tissue IHC reliability). GLRX has no transmembrane segment (UniProt P35754: topology).
| Kidney tubule cells show clear cytoplasmic chromogenic staining, with little deposit outside the cells. | This fits a useful positive control: kidney tubule cells are High by tissue IHC (HPA: kidney), and GLRX is cytoplasmic (UniProt P35754: subcellular location). Compare the stained cells and compartment together; staining intensity alone does not establish specificity (general IHC practice). |
| Signal is confined to nuclei or forms a discrete intracellular pattern without convincing cytoplasmic staining. | A predominantly nuclear pattern conflicts with the annotated cytoplasmic location (UniProt P35754: subcellular location) and HPA tissue profile (HPA: cytoplasmic expression). Reassess morphology and controls before calling it GLRX; nonspecific binding or chromogen artefact can produce misleading deposits (general IHC practice). |
| Breast adipocytes or colon endothelial cells stain strongly while expected positive cells are weak. | Those specific cell populations are Not detected in HPA tissue IHC (HPA: breast adipocytes; colon endothelial cells). Check antibody specificity and endogenous detection activity with suitable controls (general IHC practice). Their HPA result is a cell-specific comparator, not a claim that every cell in either tissue lacks GLRX. |
| A low-contrast haze covers cells, stroma, and empty spaces rather than following cell boundaries. | Uniform slide-wide deposit is difficult to reconcile with the reported cell-specific cytoplasmic profile (HPA: tissue IHC). Review the no-primary control, blocking, washes, and chromogen development (general IHC practice). The pattern cannot be assigned to GLRX from appearance alone. |
| Kidney tubule cells are unstained, including in the positive-control section. | HPA reports High staining in kidney tubule cells (HPA: kidney). First check section integrity, retrieval, antibody application, and detection using appropriate process controls (general IHC practice). HPA's Approved rating has medium RNA-staining consistency (HPA: tissue IHC reliability), so a single negative section does not establish biological absence. |
💡Expected GLRX appearanceCall a positive result when cytoplasmic staining is clear in an HPA High cell population such as kidney tubule cells or duodenal glandular cells (HPA: kidney; duodenum; UniProt P35754: cytoplasm); isolated nuclear deposit or broad cell-independent haze warrants control review (general IHC practice).
| Compartment and topology | GLRX is annotated in the cytoplasm and has no transmembrane segment (UniProt P35754: location; topology). In paraffin IHC, judge the cell-associated cytoplasmic pattern; the topology entry alone does not predict staining at a visible cell boundary or specify an antibody epitope. |
| Choice of tissue comparator | Kidney tubule, duodenal glandular, and pancreatic exocrine glandular cells are High (HPA: tissue IHC). Appendix glandular cells are Medium, while thyroid glandular cells are Low (HPA: tissue IHC). Choose a clearly annotated High population for the positive comparison, and record the exact cell type when interpreting weaker fields. |
| Strength of tissue evidence | The HPA tissue profile is Approved, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability). Three listed antibodies have Approved IHC status (HPA: antibody validation). These ratings support a reference pattern, but do not prove that every positive deposit from a separate catalog antibody is specific. |
| Protein processing and epitope | UniProt lists the GLRX chain as residues 2–106, with no signal peptide, propeptide, or annotated isoforms (UniProt P35754: processing; isoforms). It also lists modified residues at positions 2 and 9 (UniProt P35754: modified residues). Without a mapped antibody epitope, these facts do not predict retrieval needs or fixation sensitivity. |
| IF/ICC Q&A: what localisation should be expected? | HPA reports cytosol as supported and plasma membrane as approved in ICC-IF (HPA: subcellular). That observation can inform interpretation of the separate IF/ICC guide; it does not override the cytoplasmic tissue IHC profile or supply an IF/ICC protocol (HPA: tissue IHC; subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in a kidney positive-control section. | The expected High kidney tubule pattern is missing (HPA: kidney); the failed step is not identifiable from this image alone. | Confirm tubules are present, then review retrieval, antibody application, detection reagents, and control-slide results in sequence (general IHC practice). Repeat the control before interpreting test sections. |
| Every tissue compartment stains, including areas outside cells. | Widespread deposit conflicts with the cell-associated cytoplasmic pattern (HPA: tissue IHC; UniProt P35754: cytoplasm). Excess detection background is possible (general IHC practice). | Inspect no-primary controls; review blocking, wash stringency, and chromogen development (general IHC practice). Reassess specificity only after the background is controlled. |
| The signal appears mainly nuclear. | Predominant nuclear staining is inconsistent with the reported cytoplasmic location (UniProt P35754: location; HPA: tissue IHC). | Check counterstain and focus, then compare positive and no-primary controls (general IHC practice). Do not score nuclear-only staining as the expected GLRX pattern without independent validation. |
| Breast adipocytes or colon endothelial cells show unexpected strong staining. | HPA lists those cell populations as Not detected (HPA: breast adipocytes; colon endothelial cells); cross-reactivity or endogenous detection activity is possible (general IHC practice). | Verify cell identity on the section and compare no-primary and detection-system controls (general IHC practice). Report the discrepancy by cell type rather than treating the entire tissue as an HPA negative. |
| A Low population stains as strongly as the kidney control. | HPA lists thyroid glandular cells as Low and kidney tubule cells as High (HPA: thyroid gland; kidney), but staining levels are qualitative reference categories. | Compare matched sections processed together and inspect controls (general IHC practice). Record the observed intensity and cell type; avoid converting HPA categories into a fixed numeric cutoff. |
| The IHC pattern differs from an ICC-IF image showing a cell-edge signal. | HPA reports approved plasma-membrane and supported cytosolic localisation in ICC-IF (HPA: subcellular), while its tissue IHC profile describes cytoplasmic expression (HPA: tissue IHC). | Interpret each preparation against its own HPA reference and verify the IHC cell type and controls (general IHC practice). Do not treat ICC-IF cell-edge signal alone as proof of a membrane-restricted paraffin IHC pattern. |
Experiment evidence
IHC & IF images showing GLRX staining
A03392-1 · IHCIHC analysis of GLRX using anti-GLRX antibody (A03392-1). GLRX was detected in a paraffin-embedded section of human kidney tissue. Heat medi
A03392-1 · IFIF analysis of Glutaredoxin 1/GLRX using anti-Glutaredoxin 1/GLRX antibody (A03392-1). Glutaredoxin 1/GLRX was detected in an immunocytochem
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000173221-GLRX/tissue/cerebral+cortex#imid_18966945
HPA · DuodenumHuman Protein Atlas IHC, Duodenum. Source: https://www.proteinatlas.org/ENSG00000173221-GLRX/tissue/duodenum#imid_18966862
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000173221-GLRX/tissue/colon#imid_18966871
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000173221-GLRX/tissue/liver#imid_18966856
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB008634; green = GLRX, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB008634; green = GLRX, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GLRX IHC & IF
🧪Run kidney first: tubular cells should stain strongly (HPA: High in cells in tubules). Use cervix glandular cells as a negative comparator (HPA: Not detected); on the kidney slide, treat morphologically identifiable cells without staining as an internal background reference, without assuming a particular renal cell type is negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Breast (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GLRX in A-431, U-251MG, U2OS, with annotated localisation: Plasma membrane (approved), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and concentration-matched nonimmune rabbit IgG matched to the primary antibody’s format (selected-SKU caption: rabbit anti-GLRX); use validated GLRX knockout material, if available, as a biological negative. Quench endogenous peroxidase in kidney sections before HRP/DAB detection (selected-SKU caption: HRP/DAB).
⚠️Feasibility: Paraffin-section kidney IHC is demonstrated with heat-mediated EDTA retrieval at pH 8.0 (selected-SKU caption), but that single condition does not establish whether retrieval is required. A target-specific fixation window and fixation effect are unreported, and the selected-SKU caption does not report a fixative (selected-SKU caption: fixative not stated). Frozen sections and IF cannot be ranked as easier from the supplied evidence; kidney endogenous peroxidase may contribute background with HRP/DAB detection (selected-SKU caption: HRP/DAB).
HPA tissue IHC evidence for GLRX
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Pancreas | Exocrine glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Colon | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GLRX IHC Tips
Use the kidney IHC example as a starting point, then judge GLRX staining by compartment, cell type, controls and the limits of the available evidence.
Which retrieval conditions should I try first for GLRX paraffin-section IHC?
Begin with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A03392-1). The selected kidney image used this retrieval before overnight incubation at 4°C with 2 μg/ml primary antibody (datasheet A03392-1). Keep retrieval conditions identical across comparison sections, and include kidney tubules as a positive tissue reference because their staining is reported as high (HPA: kidney tubule cells, High). If staining is weak, optimize heating and cooling against preserved morphology before considering another buffer as a fallback (standard IHC practice). The image caption identifies paraffin embedding but does not report the fixative, so it cannot establish a fixation-specific retrieval requirement (datasheet A03392-1).
How should I handle fixation when GLRX staining varies between paraffin blocks?
The selected tissue image identifies a paraffin-embedded kidney section but gives no fixative or fixation duration; target-specific fixation sensitivity is therefore unknown (datasheet A03392-1). Record each block’s fixative and processing history, then compare sections processed together with the same EDTA pH 8.0 retrieval and detection conditions (datasheet A03392-1; standard IHC practice). Inspect morphology alongside staining, because damaged tissue can make differences in chromogen intensity difficult to interpret (standard IHC practice). Use kidney tubule cells as a positive reference, while recognizing that their reported high staining does not establish which fixation condition is best (HPA: kidney tubule cells, High).
Should GLRX staining appear only in the cytoplasm?
Expect a substantial cytoplasmic signal in tissue sections: UniProt assigns GLRX to the cytoplasm, and the tissue atlas describes cytoplasmic expression in several tissues, including kidney (UniProt P35754 subcellular location; HPA tissue IHC). A cell-imaging dataset also reports cytosol and plasma membrane localisation, so a membrane-associated pattern merits review rather than automatic rejection (HPA subcellular: plasma membrane approved, cytosol supported). GLRX has no annotated transmembrane segment, which does not by itself rule out membrane association (UniProt P35754 topology). Compare the distribution within intact cells against no-primary controls and adjacent morphology before scoring a sharply outlined membrane signal (standard IHC practice).
Can isoforms or processing explain inconsistent GLRX staining?
UniProt lists 0 GLRX isoforms and describes the mature chain as residues 2–106, so an annotated isoform switch does not explain differing IHC patterns (UniProt P35754 isoforms and processing). The supplied antibody caption does not identify its binding epitope; do not assume that staining measures a particular region of GLRX (datasheet A03392-1). UniProt records N-acetylalanine at residue 2 and N6-succinyllysine at residue 9, but these annotations do not establish an effect on this antibody’s binding (UniProt P35754 modified residues). When sections disagree, compare retrieval, tissue preservation and staining controls before proposing an epitope-specific mechanism (standard IHC practice).
How can I assess GLRX by multiplex IF alongside the IHC findings?
Treat IF as a separate assay and validate antibody staining with appropriate controls before comparing it with chromogenic IHC (standard IF practice). Pair GLRX with a marker that identifies the cell population being assessed; kidney tubule cells are one documented high-staining population for choosing a comparison region (HPA: kidney tubule cells, High). Select fluorophores after checking the section’s autofluorescence in each channel, and keep single-stain controls to detect spectral bleed-through (standard IF practice). For an intracellular epitope, assess whether permeabilisation is needed to expose cytosolic GLRX; do not presume the antibody epitope or its side of a membrane from the tissue-IHC caption (UniProt P35754 subcellular location; datasheet A03392-1).
What should I check when GLRX DAB staining looks diffuse?
First compare the section with a no-primary control and inspect whether diffuse color follows tissue edges, damaged areas or the expected cell boundaries (standard IHC practice). The selected protocol uses a peroxidase-linked secondary and DAB, so include an endogenous peroxidase block and check its effectiveness as part of the chromogenic workflow (datasheet A03392-1; standard IHC practice). Reassess blocking, washes and primary concentration if background persists; the kidney example used 10% goat serum and 2 μg/ml primary antibody (datasheet A03392-1). Preserve the same DAB development conditions across controls and test sections so differences in color do not merely reflect development time (standard IHC practice).
How should I score GLRX staining across kidney sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring; kidney tubule cells provide a documented high-staining population, while the tissue atlas describes GLRX expression as cytoplasmic in several tissues (HPA: kidney tubule cells, High; HPA tissue IHC). For chromogenic sections, report the percentage of positive target cells and an intensity-weighted H-score, or positive-cell density per mm² of evaluable tissue (standard IHC practice). Normalize comparisons to the number of intact target cells or evaluable target-tissue area, and exclude folds, edges and damaged regions consistently (standard IHC practice). Keep retrieval, antibody incubation and DAB development consistent, and state whether membrane-associated staining was included in the score (datasheet A03392-1; HPA subcellular; standard IHC practice).
How can I distinguish convincing GLRX staining from artefact?
A convincing result places staining in identifiable cells with a plausible intracellular distribution and reproduces it across comparable sections and controls (UniProt P35754 subcellular location; standard IHC practice). Kidney tubule cells are a useful positive reference because high staining is reported there, but staining elsewhere should be judged by its own cell type and morphology (HPA: kidney tubule cells, High; standard IHC practice). Investigate isolated nuclear-only color, edge accentuation, necrotic areas and signal reproduced without primary antibody before assigning those pixels to GLRX (UniProt P35754 subcellular location; standard IHC practice). With DAB detection, residual endogenous peroxidase can also mimic positivity; the atlas labels its tissue-IHC reliability Approved with medium staining-to-RNA consistency (datasheet A03392-1; HPA tissue IHC).
Boster reagents
Best GLRX / Glutaredoxin-1 IHC Antibodies
A03392-1 has IHC data from a human kidney paraffin section (catalog IHC caption) and IF/ICC data from A431 cells (catalog IF caption); its listed reactivity is human (catalog: Human).
Real IHC data
Anti-Glutaredoxin 1/GLRX Antibody ®
Cat # A03392-1
A03392-1 will render with human kidney paraffin-section IHC data (catalog IHC caption). The same SKU lists IF and ICC applications (catalog: applications) and shows IF staining of A431 cells (catalog IF caption).
Which to pick: Choose A03392-1 for human paraffin-section IHC: its image shows kidney tissue with EDTA heat retrieval and 2 μg/ml primary antibody (catalog IHC caption); the fixative is unreported (catalog IHC caption). For IF/ICC, the same SKU lists both applications and shows A431-cell IF at 5 μg/ml (catalog: applications; catalog IF caption). No SKU in this payload lists reactivity beyond human, so there is no supported cross-species choice (catalog: reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P35754 (GLRX1_HUMAN, Glutaredoxin-1).
- Human Protein Atlas. GLRX tissue IHC expression (reliability: Approved).
- Human Protein Atlas. GLRX subcellular location (ICC-IF): Localized to the plasma membrane and cytosol..
- Human Protein Atlas. GLRX antibody validation summary (3 antibodies).
- Functional recovery of diabetic mouse hearts by glutaredoxin-1 gene therapy: role of Akt-FoxO-signaling network. Gene therapy 2010 — PMC2851834.
- Mechanism of miR-132-3p Promoting Neuroinflammation and Dopaminergic Neurodegeneration in Parkinson's Disease. eNeuro 2022 — PMC8805200.
- NRF1 mitigates motor dysfunction and dopamine neuron degeneration in mice with Parkinson's disease by promoting GLRX m(6) A methylation through upregulation of METTL3 transcription. CNS neuroscience & therapeutics 2024 — PMC10916419.
- The anti-fibrotic drug pirfenidone inhibits liver fibrosis by targeting the small oxidoreductase glutaredoxin-1. Science advances 2021 — PMC8442864.
- PubMed PMID:8018729 — UniProt-cited evidence.
- PubMed PMID:7851394 — UniProt-cited evidence.
- PubMed PMID:8645179 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GLRX (Glutaredoxin-1). Expected localisation Cytoplasmic in tissue sections (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03392-1). Validated IHC antibody: A03392-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Breast. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human kidney tissue; fixative not specified (datasheet A03392-1); 2. Fixation: Image fixative and duration unreported (datasheet A03392-1); 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: EDTA pH 8.0 (datasheet A03392-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03392-1); 8. Primary antibody: Rabbit anti-GLRX, 2-5 μg/ml (datasheet A03392-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03392-1). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: No isoforms annotated; chain spans residues 2–106 (UniProt); Regulation: Regulation not annotated (UniProt). Evidence: UniProt P35754, HPA tissue IHC · Approved, HPA ICC-IF · plasma membrane and cytosol., Datasheet A03392-1, PMC8805200, PMC10916419. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GLRX/immunohistochemistry-ihc-guide.