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GSR / Glutathione reductase, mitochondrial · IHC design guide
Design Immunohistochemistry for GSR
Plan chromogenic IHC for GSR in paraffin sections using the catalog antibody's documented conditions (datasheet: A01479-1). Compare staining with the widespread cytoplasmic and nuclear tissue pattern (HPA tissue IHC), while treating nuclear localisation cautiously because external data contradict it (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GSR 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 and nuclear tissue staining (HPA tissue IHC); mitochondrial localisation (UniProt) | |
| Staining pattern | Widespread cytoplasmic and nuclear staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01479-1) | |
| 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 | Nuclear staining needs independent verification (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | Five isoforms; check which epitopes the antibody covers (UniProt) |
Section 1
Recommended GSR IHC & IF Protocols
Compare the catalog antibody’s EDTA pH 8.0 IHC protocol (datasheet: A01479-1) with two published mouse lung GSR protocols (PMC12133717; PMC2634824).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A01479-1) |
| Fixation | Image fixative and duration unreported (datasheet A01479-1); 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 A01479-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01479-1) |
| Primary antibody | Rabbit anti-GSR, 2-5μg/ml (datasheet A01479-1) |
| Primary incubation | Overnight at 4 °C (datasheet A01479-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01479-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GSR-positive staining in glandular cells of epididymis (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic and nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet: A01479-1); both published mouse lung protocols used citrate pH 6.0 (PMC12133717; PMC2634824).
Section 2
What Is the Expected GSR Staining Pattern?
GSR is a soluble protein associated with the cytoplasm and mitochondria, with no transmembrane segment (UniProt P00390). In paraffin sections, expect cytoplasmic staining in glandular cells of epididymis and fallopian tube, where HPA reports high staining (HPA tissue IHC). HPA also describes a ubiquitous cytoplasmic and nuclear profile, but rates its IHC evidence Approved with medium consistency and a subcellular location contradicted by external data (HPA tissue IHC).
| Glandular cells show clear cytoplasmic chromogen in epididymis or fallopian tube. | This fits the reported high staining in those cells (HPA tissue IHC) and GSR's cytoplasmic localization (UniProt P00390). Score the glandular cells themselves and compare signal with adjacent tissue background. |
| Signal is mainly nuclear, or forms a sharp outline around cells. | HPA describes nuclear staining in tissue but flags a subcellular conflict (HPA tissue IHC). A nuclear-dominant pattern alone is therefore inconclusive; a crisp cell-surface outline also conflicts with the lack of a transmembrane segment (UniProt P00390). Review controls before assigning either pattern to GSR. |
| Cells expected to be unstained show strong chromogen, especially where nearby glandular cells also stain. | Adipocytes and cardiomyocytes are reported as not detected in their respective tissues (HPA tissue IHC). Strong signal in those cells warrants checks for antibody cross-reactivity or endogenous chromogenic detection activity; a tissue-level negative call would overstate the cell-specific HPA observation. |
| Pale, diffuse color covers cells and extracellular areas with little separation between them. | Poor contrast prevents a reliable cell-level interpretation. Treat widespread haze as possible assay background, including nonspecific antibody binding or detection activity, and compare it with a section processed without primary antibody (general IHC practice). |
| No staining appears in glandular cells of an epididymis or fallopian tube section. | That result conflicts with the high staining reported for those cells (HPA tissue IHC). Check section integrity, the antibody's IHC-P instructions, detection reagents and a working positive control before interpreting other negative areas as true absence (general IHC practice). |
💡Expected GSR appearanceA convincing positive is clear cytoplasmic staining in epididymal or fallopian-tube glandular cells at the reported high level (HPA tissue IHC; UniProt P00390); nuclear-only color, sharp membrane outlines or uniform slide-wide haze needs control-based review.
| Which cells provide an interpretable contrast? | Epididymal and fallopian-tube glandular cells are high; adipocytes and cardiomyocytes are not detected in their listed tissues (HPA tissue IHC). Compare named cell types, not entire organs. |
| How should nuclear chromogen be weighed? | HPA reports cytoplasmic and nuclear tissue staining yet notes an externally contradicted subcellular location (HPA tissue IHC). UniProt lists cytoplasm and mitochondrion (UniProt P00390); nuclear-only staining needs independent support. |
| Do all GSR forms imply one fine-scale pattern? | UniProt lists mitochondrial and cytoplasmic isoforms among five forms (UniProt P00390). A cytoplasmic signal is compatible with GSR; this record does not establish which isoform an IHC antibody recognizes or whether chromogen resolves mitochondria. |
| Does the protein's topology predict a membrane rim? | No transmembrane segment is annotated (UniProt P00390). A strong, continuous cell-surface rim is therefore unexpected for GSR and should be checked against background and antibody specificity. |
| How strong is the antibody-based tissue evidence? | Two listed antibodies have Approved IHC status, while one has no listed IHC status (HPA antibodies). The tissue profile itself has medium staining–RNA consistency and pending external verification (HPA tissue IHC); interpret discordant compartments cautiously. |
| IF/ICC: what localization is reported? | HPA reports mainly cytosol, with additional endoplasmic-reticulum localization, in ICC-IF (HPA subcellular). This is context for comparing localization; IF/ICC workflow belongs in its separate guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected glandular-cell signal is absent. | A failed staining run or unsuitable working conditions are possible (general IHC practice); HPA reports high signal in these cells (HPA tissue IHC). | Confirm the named cells are present, run a known-positive section and follow the IHC-validated antibody's documented IHC-P procedure. Check detection reagents before drawing a biological conclusion (general IHC practice). |
| Nuclear signal dominates with little cytoplasmic staining. | HPA describes nuclear tissue staining but flags its subcellular assignment as contradicted by external data (HPA tissue IHC); cytoplasm and mitochondrion are listed by UniProt (UniProt P00390). | Record nuclear and cytoplasmic scores separately. Compare the pattern across positive tissues and controls; avoid calling nuclear-only signal confirmed GSR localization. |
| A uniform membrane rim or unrelated cell population stains strongly. | GSR has no annotated transmembrane segment (UniProt P00390). Antibody cross-reactivity or nonspecific detection is possible (general IHC practice). | Compare cell types with HPA's cell-specific observations, include a no-primary control and reassess specificity if the unexpected pattern persists (HPA tissue IHC; general IHC practice). |
| Brown haze obscures cell boundaries. | Nonspecific antibody binding or chromogenic detection background can reduce contrast (general IHC practice). | Inspect the no-primary control, confirm blocking and washes, and optimize antibody concentration within its documented IHC-P guidance (general IHC practice). Score only resolved cellular signal. |
| Chromogen appears even without primary antibody. | Endogenous activity or nonspecific detection-reagent binding may generate color (general IHC practice). | Address the detection system's endogenous-activity block and reagent controls, then repeat the comparison with primary antibody (general IHC practice). Do not count control-matched color as GSR. |
| A presumed negative tissue contains some positive cells. | HPA's not-detected entries name particular cells, such as adipocytes or cardiomyocytes, rather than every cell in an organ (HPA tissue IHC). | Identify and score the exact cell type before declaring a discrepancy. If the named negative cells stain, compare their intensity and distribution with positive tissue and controls (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing GSR staining
A01479-1 · IHCIHC analysis of Glutathione Reductase using anti-Glutathione Reductase antibody (A01479-1). Glutathione Reductase was detected in a paraffin
A01479-1 · IHCIHC analysis of Glutathione Reductase using anti-Glutathione Reductase antibody (A01479-1). Glutathione Reductase was detected in a paraffin
A01479-1 · IFIF analysis of Glutathione Reductase using anti-Glutathione Reductase antibody (A01479-1). Glutathione Reductase was detected in an immunocy
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000104687-GSR/tissue/cerebral+cortex#imid_891048
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000104687-GSR/tissue/colon#imid_890990
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000104687-GSR/tissue/liver#imid_890983
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000104687-GSR/tissue/kidney#imid_890984
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA064806; green = GSR, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA064806; green = GSR, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GSR IHC & IF
🧪Run epididymis first; its glandular cells should stain for GSR (HPA: High in epididymal glandular cells). Use adipose tissue adipocytes as the negative comparator (HPA: Not detected in adipocytes), and assess nonglandular cells on the epididymis slide for background signal without assuming they lack GSR.
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 GSR in A-549, Hep-G2, U2OS, A-431, U-251MG, KOLF2.1J, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host- and clonality-matched isotype controls; the caption identifies a rabbit primary but does not report clonality (selected-SKU IHC caption: rabbit primary). Use GSR knockout material as a biological negative, or cognate peptide competition if the immunizing peptide is available; for chromogenic detection, block endogenous peroxidase and check for background DAB signal (selected-SKU IHC caption: HRP/DAB detection).
⚠️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 IHC caption: fixative unreported). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that this retrieval is required; the supplied evidence also does not establish that frozen sections or IF are easier than IHC-P (selected-SKU IHC caption: EDTA retrieval; HPA: ICC-IF localization). No epididymis-specific artefact is documented in the supplied evidence; inspect gland lumens and adjacent cells for background against the controls (HPA: High in epididymal glandular cells).
HPA tissue IHC evidence for GSR
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Subcellular location is contradicted by external data. Pending external verification.). 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 → |
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | 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 → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GSR IHC Tips
Use the catalog antibody’s tissue staining conditions as the IHC starting point, then check compartment, cell type and controls before interpreting GSR signal.
Which retrieval conditions should I try first for GSR in paraffin sections?
Start with heat-mediated EDTA retrieval at pH 8.0 for paraffin sections (datasheet A01479-1). The catalog antibody stained a paraffin-embedded human lung cancer section after this retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet A01479-1). If staining is weak, adjust heating time on matched sections while keeping buffer and detection conditions constant, and compare tissue structure as well as signal (standard IHC practice). Use a no-primary control to identify detection background, and judge improvement in plausible cytoplasmic staining rather than a rise in staining across every compartment (UniProt P00390 localisation; standard IHC practice).
How should I troubleshoot fixation-related loss of GSR staining?
Target-specific fixation sensitivity is unknown: the selected tissue-IHC caption identifies a paraffin-embedded section but does not state its fixative (datasheet A01479-1). Record the actual fixative and fixation duration for each specimen, then compare similarly processed sections before changing retrieval or antibody concentration (standard IHC practice). For routine formalin-fixed material, review tissue preservation and test a small retrieval-time series with the stated EDTA pH 8.0 method; this is a general troubleshooting step, not a demonstrated GSR-specific rescue (datasheet A01479-1; standard IHC practice). Interpret apparent signal loss against a same-run positive tissue control and a no-primary control (standard IHC practice).
What GSR staining pattern is plausible, and how should I assess nuclear signal?
Assess cytoplasmic staining first: GSR is annotated in the cytoplasm and mitochondrion, while HPA subcellular data support mainly cytosolic localisation (UniProt P00390 localisation; HPA subcellular). The HPA tissue-IHC profile also describes nuclear expression, but its reliability note says subcellular location is contradicted by external data and awaits verification (HPA tissue IHC). Score nuclear staining separately from cytoplasmic staining rather than combining them into one positive result (standard IHC practice). If nuclei dominate, compare no-primary staining, retrieval intensity and a second validated approach before assigning that pattern to GSR (HPA tissue IHC reliability; standard IHC practice).
Can this antibody distinguish mitochondrial and cytoplasmic GSR isoforms in IHC?
Do not assign an IHC signal to one isoform without an epitope map and isoform-specific validation for this antibody (standard IHC practice). The record lists 5 GSR isoforms, including mitochondrial and cytoplasmic forms, and annotates a mature mitochondrial chain spanning residues 44–522 (UniProt P00390 isoforms and processing). It also lists no transmembrane segment and one modified residue, acetyllysine 97; neither annotation establishes which forms this antibody detects in fixed sections (UniProt P00390 topology and modified residues). Compare staining with independently validated isoform-specific evidence if isoform attribution is essential, while reporting routine chromogenic results as GSR immunoreactivity (standard IHC practice).
How can IF help check an ambiguous GSR IHC pattern?
Use IF as a separate validation experiment and multiplex GSR with a marker of the expected cell type, such as a validated glandular-cell marker when examining the HPA-positive epididymis or fallopian tube (HPA tissue IHC; standard IF practice). Choose fluorophores after inspecting unstained tissue for autofluorescence, and reserve a well-separated channel for the weaker signal (standard IF practice). Because GSR has cytoplasmic and mitochondrial annotations but no transmembrane segment, permeabilise sufficiently to access intracellular epitopes; the antibody’s exact epitope and membrane orientation are unreported (UniProt P00390 localisation and topology; standard IF practice). Compare compartment patterns with the chromogenic section without treating IF conditions as validated by the tissue-IHC caption (datasheet A01479-1; standard IF practice).
What should I change when GSR DAB staining is widespread or patchy?
First compare a no-primary section with the stained section to locate secondary-reagent or chromogen background (standard IHC practice). The selected example used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, a peroxidase-conjugated secondary and DAB detection (datasheet A01479-1). Apply an endogenous-peroxidase block and check wash stringency as general chromogenic IHC controls, especially before interpreting diffuse brown deposits (standard IHC practice). Because GSR has broad tissue expression and HPA reports a ubiquitous cytoplasmic and nuclear IHC profile, background must be judged against cell morphology, compartment and controls rather than staining prevalence alone (UniProt P00390 localisation; HPA tissue IHC).
How should I quantify GSR IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic compartment before scoring, and keep nuclear staining as a separate readout because its interpretation is disputed (UniProt P00390 localisation; HPA tissue IHC reliability). For cellular staining, report the percentage of positive cells and an H-score using fixed intensity categories; retain the same threshold and imaging settings across the comparison (standard IHC practice). If the question concerns infiltrating positive cells, report positive-cell density per mm² of viable tissue rather than total brown area (standard IHC practice). Normalise each result to the eligible cell count or viable tissue area, exclude necrosis and section edges, and include same-run controls (standard IHC practice).
How do I separate genuine GSR staining from artefact?
A credible result shows staining in intact cells with a plausible cytoplasmic distribution, consistent with GSR’s cytoplasmic and mitochondrial annotation and HPA’s mainly cytosolic subcellular result (UniProt P00390 localisation; HPA subcellular). Cell identity matters: HPA reports high staining in epididymal and fallopian-tube glandular cells, while several listed cell populations are not detected (HPA tissue IHC). Treat isolated nuclear dominance cautiously because HPA flags a subcellular discrepancy; examine edge staining, necrotic deposits and staining on no-primary sections before calling a positive result (HPA tissue IHC reliability; standard IHC practice). Investigate residual brown signal after omitting primary antibody or peroxidase blocking as possible endogenous enzyme or detection artefact (standard IHC practice).
Boster reagents
Best GSR / Glutathione reductase, mitochondrial IHC Antibodies
The IHC-validated anti-GSR antibody has paraffin-section images from human lung, tonsil, rectal, and thyroid samples (catalog IHC captions) and an IF image from A549 cells (catalog IF caption).
Real IHC data
Anti-Glutathione Reductase/GSR Antibody ®
Cat # A01479-1
A01479-1 has IHC images from paraffin sections of human lung cancer, tonsil, rectal cancer, and thyroid cancer (catalog IHC captions). Its IF/ICC image uses A549 cells (catalog IF caption), and its listed reactivity is Human, Mouse, and Rat (catalog applications and reactivity).
Which to pick: Choose A01479-1 for tissue IHC: its human paraffin-section images used EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (catalog IHC captions); the fixative is unreported (catalog IHC captions). For IF/ICC, A01479-1 has an A549 cell image using 5 μg/ml primary antibody (catalog IF caption). For mouse or rat samples, A01479-1 lists both species as reactive, while the supplied IHC images show human samples (catalog reactivity; catalog IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P00390 (GSHR_HUMAN, Glutathione reductase, mitochondrial).
- Human Protein Atlas. GSR tissue IHC expression (reliability: Approved).
- Human Protein Atlas. GSR subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the endoplasmic reticulum..
- Human Protein Atlas. GSR antibody validation summary (3 antibodies).
- Distinct roles for the thioredoxin and glutathione antioxidant systems in Nrf2-Mediated lung tumor initiation and progression. Redox biology 2025 — PMC12133717.
- A Novel Biological Activity of the STAT3 Inhibitor Stattic in Inhibiting Glutathione Reductase and Suppressing the Tumorigenicity of Human Cervical Cancer Cells via a ROS-Dependent Pathway. OncoTargets and therapy 2021 — PMC8275107.
- Identification of Metabolic-Associated Genes for the Prediction of Colon and Rectal Adenocarcinoma. OncoTargets and therapy 2021 — PMC8020594.
- Nrf2-regulated glutathione recycling independent of biosynthesis is critical for cell survival during oxidative stress. Free radical biology & medicine 2009 — PMC2634824.
- PubMed PMID:2185014 — UniProt-cited evidence.
- PubMed PMID:10708558 — UniProt-cited evidence.
- PubMed PMID:20628807 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GSR (Glutathione reductase, mitochondrial). Expected localisation Cytoplasmic and nuclear tissue staining (HPA tissue IHC); mitochondrial localisation (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01479-1). Validated IHC antibody: A01479-1 at 2-5μg/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 A01479-1); 2. Fixation: Image fixative and duration unreported (datasheet A01479-1); 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 A01479-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01479-1); 8. Primary antibody: Rabbit anti-GSR, 2-5μg/ml (datasheet A01479-1); 9. Primary incubation: Overnight at 4 °C (datasheet A01479-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Nuclear staining needs independent verification (HPA tissue IHC); Isoform / epitope: Five isoforms; check which epitopes the antibody covers (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt P00390, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A01479-1, PMC12133717, PMC2634824. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GSR/immunohistochemistry-ihc-guide.