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GLUD2 / Glutamate dehydrogenase 2, mitochondrial · IHC design guide
Design Immunohistochemistry for GLUD2
Plan GLUD2 chromogenic IHC on paraffin sections using catalog antibody A04531 (datasheet A04531). Interpret variable cytoplasmic staining (HPA tissue IHC) alongside mitochondrial matrix localization (UniProt), with caution because the tissue staining may detect protein from more than one gene (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GLUD2 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 | Variable cytoplasmic staining (HPA tissue IHC); mitochondrial matrix localization (UniProt) | |
| Staining pattern | Variable cytoplasmic staining in glandular and glial cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, heat-mediated (datasheet A04531) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Staining may detect protein from more than one gene (HPA tissue IHC) | |
| Regulation | No staining-linked regulator reported (UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain spans residues 54–558 (UniProt) |
Section 1
Recommended GLUD2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A04531) is paired with a published protocol for FFPE glioblastoma tissues (PMC6284416).
| Sample | Paraffin-embedded human colon carcinoma tissue; fixative not specified (datasheet A04531) |
| Fixation | Image fixative and duration unreported (datasheet A04531); 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 6.0 (datasheet A04531); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-GLUD2, 1:50-1:200 (datasheet A04531) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GLUD2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression at variable levels. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate pH 6.0 retrieval (datasheet A04531); the published protocol used MS-unmasker solution in a microwave (PMC6284416).
Section 2
What Is the Expected GLUD2 Staining Pattern?
GLUD2 is a mitochondrial matrix protein without a transmembrane segment (UniProt P49448). In IHC, assess cytoplasmic staining in the cell types HPA reports as positive, including glandular cells and caudate glial cells (HPA tissue IHC). Interpret apparent positives cautiously: HPA rates tissue IHC Uncertain because staining agrees poorly with RNA data and antibodies may detect proteins from more than one gene (HPA tissue IHC).
| Cytoplasmic staining is strongest in adrenal or appendix glandular cells, or bronchial respiratory epithelial cells (HPA: High). | This matches reported high-staining cells, but agreement alone does not establish GLUD2 specificity (HPA tissue IHC: Uncertain). Assess the distribution within cells; GLUD2 is assigned to the mitochondrial matrix (UniProt P49448). |
| Staining is mainly nuclear or extracellular, with little cytoplasmic signal. | That distribution conflicts with mitochondrial matrix localisation (UniProt P49448; HPA subcellular: mitochondria supported). Treat it as possible artefact or nonspecific detection, and review the negative control and staining distribution before scoring it positive (general IHC practice). |
| Strong staining appears in skeletal myocytes, soft-tissue fibroblasts or bone-marrow hematopoietic cells (HPA: Low). | This contrasts with HPA's low-staining examples (HPA tissue IHC). Cross-reactivity or endogenous detection activity is possible; neither can be diagnosed from location alone. Compare controls and the expected cell distribution before assigning GLUD2 positivity (general IHC practice). |
| Colour is diffuse across cells or tissue, without a discernible cellular pattern. | Diffuse background cannot reliably identify the mitochondrial matrix (UniProt P49448). Check the no-primary control for detection background, then review blocking, washing and detection conditions as general IHC variables (general IHC practice). |
| No signal appears in an adrenal or appendix section where HPA reports high glandular-cell staining (HPA: High). | First check tissue integrity, the assay control and whether the expected cells are present (general IHC practice). A negative result merits investigation, but HPA's Uncertain IHC reliability prevents treating one section as a definitive GLUD2-negative or assay-failure verdict (HPA tissue IHC). |
💡Expected GLUD2 appearanceCall positive a cell-associated cytoplasmic signal in HPA-reported high-staining cell types, potentially punctate in keeping with mitochondrial localisation; predominantly nuclear, extracellular or uniform background colour is suspect (HPA tissue IHC; UniProt P49448; general IHC practice).
| Compartment and topology | GLUD2 is assigned to the mitochondrial matrix and has no transmembrane segment (UniProt P49448). Interpret cytoplasmic cellular signal in that context; nuclear or extracellular colour needs separate scrutiny (general IHC practice). |
| Protein processing | UniProt annotates the mature chain as residues 54–558 (UniProt P49448). If an antibody's epitope is known, compare it with that annotation; no epitope is supplied here, so no effect on staining can be predicted. |
| Tissue and cell context | HPA reports high staining in several glandular and epithelial populations and caudate glial cells, but low staining in skeletal myocytes, fibroblasts and bone-marrow hematopoietic cells (HPA tissue IHC). Use these as comparisons, not absolute controls. |
| IHC specificity evidence | HPA rates tissue IHC Uncertain, citing poor staining–RNA consistency and possible detection of proteins from more than one gene; its listed IHC antibodies are also rated Uncertain (HPA tissue IHC; HPA antibodies). |
| IF/ICC localisation evidence | HPA reports supported mitochondrial ICC-IF localisation, while warning that the antibodies may target proteins from multiple genes (HPA subcellular). That supports a compartment comparison; it does not validate a chromogenic IHC result or resolve GLUD2 specificity. |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected glandular cells show no colour in an HPA high-staining tissue (HPA tissue IHC). | The expected cells may be absent from the section, or the IHC run may have failed (general IHC practice). HPA's uncertain tissue evidence also limits this comparison (HPA tissue IHC). | Confirm the cell population and section quality, inspect the run control, then review retrieval and antibody conditions as general IHC variables. No GLUD2-specific retrieval or fixation effect is established by these sources. |
| Every region, including low-staining comparison cells, appears strongly coloured (HPA: Low examples). | Widespread detection background, endogenous activity or cross-reactivity is possible (general IHC practice); HPA also flags possible detection of proteins from more than one gene (HPA tissue IHC). | Compare the no-primary control, inspect background outside target cells and review blocking and detection conditions (general IHC practice). Avoid scoring uniform colour as GLUD2-specific. |
| Colour concentrates in nuclei or outside cells. | The distribution conflicts with mitochondrial matrix localisation (UniProt P49448; HPA subcellular: mitochondria supported). | Review control staining and cellular boundaries; score the discordant compartment separately from cytoplasmic signal (general IHC practice). Do not infer GLUD2 localisation from the discordant colour. |
| High-staining cells vary across sections of the same tissue (HPA tissue IHC). | Different sampled cell populations or variable staining may explain the difference (general IHC practice). HPA reports variable general cytoplasmic expression and Uncertain tissue IHC reliability (HPA tissue IHC). | Compare the same cell type and tissue region across sections, document the distribution and retain the uncertainty in the interpretation (general IHC practice; HPA tissue IHC). |
| The positive signal is faint and difficult to assign to individual cells. | Weak colour or counterstain may obscure cellular boundaries (general IHC practice). A fine mitochondrial pattern may be difficult to resolve by chromogenic IHC (general IHC practice). | Review the unstained and negative-control areas, then assess counterstain and detection conditions on the IHC run (general IHC practice). Report only clearly cell-associated signal; do not require visible individual mitochondria. |
| Does mitochondrial IF/ICC staining confirm the IHC call? | HPA supports mitochondrial ICC-IF localisation but cautions that the antibodies may target proteins from multiple genes (HPA subcellular). Its tissue IHC assessment remains Uncertain (HPA tissue IHC). | Use IF/ICC as a localisation cross-check only. Judge the paraffin-section result using its own cellular pattern and controls (general IHC practice); do not transfer ICC-IF support into an IHC specificity claim. |
Experiment evidence
IHC & IF images showing GLUD2 staining
A04531 · IHCImmunohistochemistry of paraffin-embedded human colon carcinoma using GLUD2 Rabbit pAb at dilution of 1:50 .Perform high pressure antigen re
A04531 · IHCImmunohistochemistry of paraffin-embedded rat kidney using GLUD2 Rabbit pAb at dilution of 1:50 .Perform high pressure antigen retrieval wit
A04531 · IFImmunofluorescence analysis of HepG2 cells using GLUD2 Rabbit pAb at dilution of 1:50 . Blue: DAPI for nuclear staining.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000182890-GLUD2/tissue/cerebral+cortex#imid_12548657
HPA · StomachHuman Protein Atlas IHC, Stomach. Source: https://www.proteinatlas.org/ENSG00000182890-GLUD2/tissue/stomach+1#imid_12548573
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000182890-GLUD2/tissue/colon#imid_12548591
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000182890-GLUD2/tissue/liver#imid_12548592
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA044839; green = GLUD2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA044839; green = GLUD2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GLUD2 IHC & IF
🧪Run caudate first: glial cells should stain (HPA: High in caudate glial cells). HPA detects GLUD2 in all 45 scored tissues (HPA: no negative tissue rows), so use no-primary and isotype controls for the negative; neighboring cells without signal should show only background staining, but the supplied HPA rows do not establish a truly GLUD2-negative cell type.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: GLUD2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GLUD2 in A-431, U-251MG, U2OS, CACO-2, Hep-G2, KOLF2.1J, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, host-matched nonimmune rabbit IgG as the polyclonal isotype control (catalog caption: rabbit pAb), and GLUD2 knockout tissue or a peptide-block control as a biological negative. For chromogenic IHC, quench endogenous peroxidase and check for residual pigment in caudate sections; for IF, assess tissue autofluorescence.
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the selected A04531 paraffin-section caption leaves the fixative unreported. That caption specifies high-pressure retrieval in 10 mM citrate, pH 6.0, before IHC at 1:50 (A04531 tissue-IHC caption); it does not establish that retrieval is required for every antibody or specimen. Frozen sections and IF have no matched protocol here, so neither can be judged easier; brain pigment and autofluorescence can complicate interpretation (standard IHC/IF practice).
HPA tissue IHC evidence for GLUD2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Very low consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). 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 → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: GLUD2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced GLUD2 IHC Tips
Use compartment-aware controls when optimizing chromogenic GLUD2 IHC: GLUD2 is a mitochondrial matrix protein, while available tissue staining has specificity limits (UniProt P49448; HPA tissue IHC).
What retrieval should I try first for weak GLUD2 staining in paraffin sections?
Begin with heat-mediated retrieval in 10 mM citrate, pH 6.0, for paraffin-section GLUD2 IHC (datasheet A04531). The selected image used high-pressure retrieval and a 1:50 primary dilution in human colon carcinoma; its caption does not report the fixative (caption A04531). If signal is weak, compare heating intervals on adjacent sections while keeping antibody dilution and detection constant; test another buffer only as a controlled fallback (standard IHC practice). Judge improvement by staining in the cytoplasm rather than nuclei or extracellular material, because GLUD2 resides in the mitochondrial matrix (UniProt P49448).
How should I investigate inconsistent GLUD2 staining between paraffin blocks?
Target-specific fixation sensitivity for GLUD2 IHC is unknown from the supplied evidence, and the A04531 paraffin caption does not state a fixative (caption A04531). Record each block’s fixative, fixation duration, processing history and section age before comparing intensity (standard IHC practice). Stain adjacent sections in one run with the same 10 mM citrate, pH 6.0 retrieval and antibody dilution so that processing differences are easier to assess (datasheet A04531; standard IHC practice). Include a known staining control section in every run, but do not assign a GLUD2-specific fixation effect from staining differences alone (standard IHC practice).
Does diffuse or nuclear chromogen support a GLUD2-positive call?
GLUD2 is assigned to the mitochondrial matrix and has no transmembrane segment, so evaluate cytoplasmic staining for a mitochondrial pattern rather than calling nuclear or membrane staining positive (UniProt P49448). HPA reports general cytoplasmic tissue staining but rates its IHC reliability uncertain because staining and RNA expression show very low consistency (HPA tissue IHC). Compare matched positive and negative control sections at the same magnification and exposure to distinguish fine cytoplasmic signal from diffuse chromogen haze (standard IHC practice). A mitochondrial counterstain on a separately prepared section can support compartment assignment, but spatial agreement alone cannot establish GLUD2 specificity (standard IHC practice).
How can I assess whether staining reflects GLUD2 rather than a related antigen?
The supplied record annotates 0 isoforms and a processed chain spanning residues 54–558, but it does not define this antibody’s epitope (UniProt P49448). Accordingly, neither retrieval response nor the presence of an ADP-ribosylcysteine at residue 172 establishes which sequence the antibody recognizes (UniProt P49448; standard IHC practice). HPA explicitly cautions that its tissue and subcellular antibodies target proteins from more than one gene, so those patterns cannot establish GLUD2-specific staining (HPA tissue IHC; HPA subcellular). Check the catalog antibody’s immunogen and cross-reactivity documentation, then use an independently validated specificity control before interpreting a positive section as GLUD2 (standard IHC practice).
How should I evaluate GLUD2 in a multiplex IF experiment?
Treat IF as a separate assay from the A04531 paraffin IHC image; that caption supplies no IF fixation or permeabilisation evidence (caption A04531). In caudate, pair GLUD2 with a validated glial marker because HPA reports high staining in glial cells there, while retaining its uncertainty warning (HPA tissue IHC). Choose a far-red GLUD2 fluorophore and inspect unstained tissue for autofluorescence before assigning puncta to signal (standard IF practice). Because GLUD2 is in the mitochondrial matrix, pilot detergent permeabilisation that permits antibody access across mitochondrial membranes, and compare with a mitochondrial marker and appropriate single-label controls (UniProt P49448; standard IF practice).
What should I check when GLUD2 chromogenic IHC has widespread brown background?
Run a no-primary control and inspect pigment, section folds and tissue edges before increasing stringency (standard IHC practice). For peroxidase-based detection, verify the endogenous peroxidase block and compare DAB development times on matched sections; these are general workflow controls, not GLUD2-specific evidence (standard IHC practice). Reduce primary concentration or incubation time one variable at a time, using the image’s 1:50 dilution only as an A04531 reference condition (caption A04531; standard IHC practice). Interpret widespread cytoplasmic staining cautiously because HPA describes variable general cytoplasmic expression and rates its tissue IHC reliability uncertain (HPA tissue IHC).
How should I score GLUD2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population, compartment and positivity threshold before scoring, and apply them consistently to all sections (standard IHC practice). For chromogenic IHC, record the percentage of positive cells and an intensity-weighted H-score from 0–300; alternatively, report positive-cell density per mm² within a predefined region (standard IHC practice). Normalize each result to the number of eligible cells or measured tissue area, and exclude folds, necrosis and section edges using prespecified rules (standard IHC practice). Report controls and staining batches alongside scores because HPA rates its GLUD2 tissue staining uncertain and warns that its antibodies can detect proteins from multiple genes (HPA tissue IHC).
Which staining features would make a GLUD2-positive result doubtful?
A plausible GLUD2 signal should occupy cytoplasm with a mitochondrial distribution, consistent with its matrix localisation (UniProt P49448). Nuclear, surface-only or extracellular staining, edge-restricted color, necrotic deposits and signal persisting in a no-primary control call for investigation before scoring (UniProt P49448; standard IHC practice). Check endogenous peroxidase and pigment when brown deposits appear with DAB detection, then compare clean regions and control sections processed in the same run (standard IHC practice). Cell identity also matters: HPA reports high caudate glial staining, yet its tissue IHC is uncertain and may detect proteins from multiple genes, so corroborate unexpected positive cells independently (HPA tissue IHC).
Boster reagents
Best GLUD2 / Glutamate dehydrogenase 2, mitochondrial IHC Antibodies
Anti-GLUD2 A04531 has IHC images from paraffin-embedded human colon carcinoma and rat kidney, plus an IF image from HepG2 cells (A04531 image captions).
Real IHC data
Anti-GLUD2 Antibody
Cat # A04531
A04531 is listed for IHC, IF and ICC in human, mouse and rat samples (catalog applications and reactivity). Its IHC captions show human colon carcinoma and rat kidney; its IF caption shows HepG2 cells (A04531 image captions).
Which to pick: For tissue IHC, choose A04531: its rabbit polyclonal antibody was imaged on paraffin-embedded sections at 1:50 after high-pressure retrieval in 10 mM citrate buffer, pH 6.0 (A04531 IHC captions); the fixative is unreported (A04531 IHC captions). For IF/ICC, A04531 is listed for both applications, with an IF image from HepG2 cells at 1:50 (catalog applications; A04531 IF caption). For cross-species work, A04531 lists human, mouse and rat reactivity, with IHC images for human and rat samples (catalog reactivity; A04531 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P49448 (DHE4_HUMAN, Glutamate dehydrogenase 2, mitochondrial).
- Human Protein Atlas. GLUD2 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. GLUD2 subcellular location (ICC-IF): Localized to the mitochondria. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. GLUD2 antibody validation summary (3 antibodies).
- Mitochondrial enzyme GLUD2 plays a critical role in glioblastoma progression. EBioMedicine 2018 — PMC6284416.
- A new mouse allele of glutamate receptor delta 2 with cerebellar atrophy and progressive ataxia. PloS one 2014 — PMC4176021.
- Disruption of cerebellar microzonal organization in GluD2 (GluRδ2) knockout mouse. Frontiers in neural circuits 2013 — PMC3747314.
- C1ql1-Bai3 signaling is necessary for climbing fiber synapse formation in mature Purkinje cells in coordination with neuronal activity. Molecular brain 2023 — PMC10367388.
- PubMed PMID:8207021 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15772651 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GLUD2 (Glutamate dehydrogenase 2, mitochondrial). Expected localisation Variable cytoplasmic staining (HPA tissue IHC); mitochondrial matrix localization (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, heat-mediated (datasheet A04531). Validated IHC antibody: A04531 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon carcinoma tissue; fixative not specified (datasheet A04531); 2. Fixation: Image fixative and duration unreported (datasheet A04531); 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 6.0 (datasheet A04531); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-GLUD2, 1:50-1:200 (datasheet A04531); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Staining may detect protein from more than one gene (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain spans residues 54–558 (UniProt); Regulation: No staining-linked regulator reported (UniProt). Evidence: UniProt P49448, HPA tissue IHC · Uncertain, HPA ICC-IF · mitochondria. Caution: Based on antibodies targeting proteins from…, Datasheet A04531, PMC6284416. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GLUD2/immunohistochemistry-ihc-guide.