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GDAP1 / Ganglioside-induced differentiation-associated protein 1 · IHC design guide
Design Immunohistochemistry for GDAP1
Plan GDAP1 chromogenic IHC in paraffin sections around cytoplasmic staining in CNS neuropil and neuronal cells (HPA tissue IHC). Cerebellar Purkinje cells and kidney distal tubules offer high-staining controls (HPA tissue IHC); the catalog antibody’s IHC range is 1:50–1:200 (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GDAP1 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 IHC (HPA tissue IHC); mitochondrial outer membrane expected (UniProt) | |
| Staining pattern | CNS neuropil and neuronal cytoplasm predominate (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Cerebellum+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 | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | No staining regulator reported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope effects are unknown (UniProt) |
Section 1
Recommended GDAP1 IHC & IF Protocols
The catalog antibody's IHC-P protocol (datasheet) is accompanied by a published immunoperoxidase protocol for mouse and rat nervous tissue (PMC2570022).
| Sample | Paraffin-embedded rat kidney tissue; fixative not specified (datasheet A03114) |
| Fixation | Image fixative and duration unreported (datasheet A03114); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| 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-GDAP1, 1:50-1:200 (datasheet A03114) |
| 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 | GDAP1-positive staining in purkinje cells - cytoplasm/membrane of cerebellum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues, most abundant in neuropil and neuronal cells of the CNS. No signal in the no-primary control. |
💡Decision noteFor paraffin sections, start with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen-retrieval rule). The published frozen-section protocol uses cold acetone (PMC2570022).
Section 2
What Is the Expected GDAP1 Staining Pattern?
GDAP1 is an outer mitochondrial membrane protein with two transmembrane segments, and UniProt also lists cytoplasmic localization (UniProt Q8TB36 topology and subcellular location). In paraffin-section IHC, expect cytoplasmic staining in neuronal cells and neuropil, with strong staining reported in cerebellar Purkinje cells and kidney distal tubules (HPA tissue IHC). HPA rates the tissue pattern Enhanced, while reporting medium consistency between antibody staining and RNA expression (HPA tissue IHC reliability).
| Purkinje cell cytoplasm or membrane and kidney distal tubules stain strongly. | These match HPA's High observations (HPA tissue IHC). Compare the stained cells with nearby tissue before judging an entire section positive; GDAP1 staining varies by cell type (HPA tissue IHC). |
| Neuronal cells or neuropil stain at moderate intensity. | Medium staining is reported in caudate and hippocampal neuronal cells and cerebral cortex neuropil (HPA tissue IHC). A weaker result than Purkinje cells can therefore still fit the observed pattern (HPA tissue IHC). |
| Nuclear staining dominates while the expected cytoplasmic pattern is absent. | Treat this as suspect localization: UniProt places GDAP1 at the outer mitochondrial membrane and in cytoplasm, and HPA describes tissue staining as cytoplasmic (UniProt Q8TB36 subcellular location; HPA tissue IHC). |
| Strong staining appears in adipocytes or other cells HPA reports as undetected. | For adipocytes, this conflicts with HPA's Not detected observations (HPA tissue IHC). Check whether staining follows cell boundaries or background; cross-reactivity or endogenous chromogenic activity are possible explanations (general IHC practice). |
| Color appears broadly across cells and extracellular areas, without a discernible cell pattern. | This does not resemble HPA's cell-specific observations (HPA tissue IHC). Diffuse chromogen can reflect nonspecific antibody binding or detection background; inspect a no-primary control before assigning GDAP1 expression (general IHC practice). |
💡Expected GDAP1 appearanceCall a section positive when cell-associated cytoplasmic staining is evident in the expected cells, especially High Purkinje cell cytoplasm or membrane or kidney distal tubules (HPA tissue IHC); dominant nuclear or diffuse extracellular color is suspect against the reported localization (UniProt Q8TB36 subcellular location; HPA tissue IHC).
| Compartment and topology | Two transmembrane segments at residues 292–312 and 320–340 anchor GDAP1 to the outer mitochondrial membrane (UniProt Q8TB36 topology and subcellular location). Interpret visible IHC color at the cellular level; the payload does not establish that individual mitochondria resolve in chromogenic sections. |
| Tissue and cell selection | Purkinje cells and kidney distal tubules are High; caudate and hippocampal neurons and cerebral cortex neuropil are Medium (HPA tissue IHC). Adipocytes and several listed glandular or epithelial populations are Not detected (HPA tissue IHC). Choose controls by the annotated cell population, not the organ name alone. |
| Strength of tissue evidence | HPA calls the tissue IHC profile Enhanced but describes only medium consistency with RNA expression (HPA tissue IHC reliability). Both listed rabbit polyclonal antibodies have Enhanced IHC validation (HPA antibodies). These ratings support comparison with the reported pattern; they do not verify every new antibody or staining run. |
| Isoforms and epitope | UniProt lists 2 GDAP1 isoforms and GST domains at residues 24–105 and 153–309 (UniProt Q8TB36 isoforms and domains). No epitope location is supplied, so isoform recognition and effects of antigen retrieval cannot be predicted from this record. |
| What should IF/ICC show? | Predominantly mitochondrial signal with additional cytosolic signal is the HPA ICC-IF observation (HPA subcellular ICC-IF). HPA lists HPA014266 as Supported for ICC (HPA antibodies). This is a localization comparison only; no IF/ICC protocol is supplied here. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in Purkinje cells or kidney distal tubules. | These are High reference populations (HPA tissue IHC); absent signal may reflect a failed stain or insufficient assay sensitivity (general IHC practice). | Confirm that the expected cells are present, inspect a concurrent positive control, and check primary-antibody and detection steps (general IHC practice). Do not infer GDAP1 absence from this slide alone. |
| Predominantly nuclear color obscures the cytoplasm. | Nuclear dominance conflicts with the reported mitochondrial and cytoplasmic locations (UniProt Q8TB36 subcellular location; HPA tissue IHC). Nonspecific binding or detection background is possible (general IHC practice). | Compare a no-primary control and assess whether the color follows nuclei across unrelated cell types (general IHC practice). Reassess specificity before scoring GDAP1. |
| Strong color occurs in adipocytes. | HPA reports adipocytes as Not detected in adipose tissue and breast (HPA tissue IHC). Cross-reactivity or background may account for an unexpected result (general IHC practice). | Check a no-primary control and compare cell-specific staining with a known-positive population on the same run (general IHC practice). Treat discordant adipocyte color cautiously. |
| Diffuse color covers tissue and empty spaces. | A widespread pattern does not match HPA's cell-specific staining observations (HPA tissue IHC). Nonspecific binding or chromogenic detection background is possible (general IHC practice). | Inspect no-primary and detection controls; review blocking, washes, and chromogen development as general IHC checks (general IHC practice). Score only a resolved cellular pattern. |
| Moderate brain staining is mistaken for a failed assay. | Caudate and hippocampal neuronal cells and cerebral cortex neuropil are Medium, whereas Purkinje cells are High (HPA tissue IHC). | Compare like cell populations and retain the HPA intensity distinctions when scoring (HPA tissue IHC). Use a High reference population to assess whether the run produced detectable signal. |
| Retrieval changes the result between runs. | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. | Record retrieval conditions and compare runs with the same positive and no-primary controls (general IHC practice). Avoid attributing a change specifically to GDAP1 epitope exposure without further evidence. |
Experiment evidence
IHC & IF images showing GDAP1 staining
A03114 · IHCImmunohistochemistry of paraffin-embedded rat kidney using GDAP1 antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10
A03114 · IHCImmunohistochemistry of paraffin-embedded rat heart using GDAP1 antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10 m
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000104381-GDAP1/tissue/cerebral+cortex#imid_6367912
HPA · CerebellumHuman Protein Atlas IHC, Cerebellum. Source: https://www.proteinatlas.org/ENSG00000104381-GDAP1/tissue/cerebellum#imid_6367915
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000104381-GDAP1/tissue/colon#imid_6370206
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000104381-GDAP1/tissue/liver#imid_6370207
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA014266; green = GDAP1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA014266; green = GDAP1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GDAP1 IHC & IF
🧪Run cerebellum first: Purkinje cells should show cytoplasmic/membrane staining (HPA: High in cerebellar Purkinje cells). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes), and assess background in adjacent cells on the cerebellum slide without assuming those cells are GDAP1-negative.
Positive control tissue: Cerebellum (Purkinje cells - cytoplasm/membrane, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GDAP1 in A-549, PC-3, U2OS, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) slide, a concentration-matched isotype control appropriate to the primary antibody’s host species and clonality, and GDAP1-knockout tissue if available (standard IHC practice). Quench endogenous peroxidase and check the cerebellum slide for pigment that could be mistaken for chromogenic signal (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence; the selected A03114 rat-kidney paraffin-section caption does not state its fixative (caption: fixative unreported). That caption uses microwave retrieval in 10 mM PBS, pH 7.2, at 1:100, but does not establish that retrieval is required for cerebellum (A03114 tissue-IHC caption). GDAP1 has a supported mitochondrial location in ICC-IF images, which can aid localisation checks, but the supplied evidence does not establish whether IF or frozen sections are easier; check neuronal pigment when interpreting cerebellar staining (HPA: subcellular localisation; standard IHC practice).
HPA tissue IHC evidence for GDAP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Kidney | Distal tubules | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuropil | Medium | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GDAP1 IHC Tips
Troubleshoot GDAP1 staining in paraffin sections by checking retrieval, compartment, cell type, controls, and scoring before interpreting differences.
Which retrieval conditions should I try first for weak GDAP1 staining?
Start GDAP1 retrieval in paraffin sections with microwave heating in 10 mM PBS, pH 7.2 (A03114 tissue-IHC caption). The caption documents staining of rat kidney at 1:100 but gives no heating time or temperature, so record those settings when reproducing the method (A03114 tissue-IHC caption). If staining remains weak, compare adjacent sections using citrate pH 6.0 at 95–98 °C for 20 min as a fallback (page retrieval rule). Keep detection and imaging conditions matched across the comparison (standard IHC practice). Check whether signal improves in distal tubules without a parallel rise in diffuse background (HPA: High in kidney distal tubules; standard IHC practice).
Could fixation explain inconsistent GDAP1 staining between paraffin blocks?
Target-specific GDAP1 sensitivity to fixation is unknown because the selected paraffin-section caption does not state its fixative (A03114 tissue-IHC caption). Record each block's fixative, fixation duration, processing history, and section age before comparing staining (standard IHC practice). For a controlled comparison, stain adjacent sections from each block in the same run with the same retrieval and antibody conditions (standard IHC practice). Include a known staining reference, such as kidney distal tubules, while recognising that its HPA pattern does not establish fixation tolerance (HPA: High in kidney distal tubules). If results differ, report the processing difference as a possible confounder, without assigning a GDAP1-specific mechanism (standard IHC interpretation).
What staining pattern fits GDAP1 in tissue sections?
Expect predominantly cytoplasmic staining, potentially with a punctate mitochondrial pattern, because GDAP1 is assigned to the mitochondrial outer membrane and cytoplasm (UniProt Q8TB36 localisation). HPA tissue staining is most abundant in CNS neuropil and neuronal cells, with High staining in Purkinje-cell cytoplasm or membrane and kidney distal tubules (HPA tissue IHC). Judge the pattern within identified cells rather than treating all brown tissue regions as equivalent (standard IHC practice). A diffuse nuclear-only pattern conflicts with the supplied localisation evidence and warrants review of controls and counterstain (UniProt Q8TB36 localisation; standard IHC practice). Bright staining alone cannot establish mitochondrial identity at chromogenic light-microscope resolution (standard IHC interpretation).
How do GDAP1 isoforms and membrane topology affect epitope interpretation?
GDAP1 has 2 listed isoforms and transmembrane segments at residues 292–312 and 320–340 (UniProt Q8TB36 isoforms and topology). The supplied product caption does not locate the antibody epitope or establish whether staining distinguishes those isoforms (A03114 tissue-IHC caption). Check the antibody's documented immunogen or epitope before assigning a signal to either isoform (standard IHC interpretation). Retrieval and section processing can alter epitope accessibility, so compare conditions while keeping antibody concentration and detection constant (standard IHC practice). Do not interpret compartmental staining as proof of cleavage: the record describes a 1–358 chain without a signal peptide or propeptide (UniProt Q8TB36 processing).
How should I validate a GDAP1 immunofluorescence result alongside IHC?
Treat IF/ICC as a separate application: the selected antibody evidence here documents paraffin-section chromogenic IHC, while HPA reports mitochondrial and additional cytosolic GDAP1 localisation by ICC/IF (A03114 tissue-IHC caption; HPA subcellular). For multiplex IF, pair GDAP1 with a marker that identifies the expected cells, such as neuronal cells or kidney distal tubules, and verify marker specificity independently (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores after checking the specimen's autofluorescence, particularly in the channel used for weak GDAP1 signal (standard IF practice). Select permeabilisation according to the documented epitope's side of the outer mitochondrial membrane; that epitope orientation is not supplied here (UniProt Q8TB36 topology; A03114 tissue-IHC caption).
What should I check when GDAP1 chromogenic staining looks diffuse?
First inspect a no-primary control for staining from the detection system and compare it with the GDAP1 section (standard IHC practice). In a peroxidase and DAB workflow, block endogenous peroxidase and check whether pigment or endogenous enzyme mimics brown product (standard chromogenic IHC practice). Reduce nonspecific staining by testing a stronger protein block, more thorough washes, or a lower antibody concentration around the caption's 1:100 starting point (A03114 tissue-IHC caption; standard IHC practice). Keep section thickness, development time, and counterstain consistent across comparisons (standard IHC practice). Prefer a cell-associated cytoplasmic pattern over uniform stromal colour when evaluating specificity (UniProt Q8TB36 localisation; standard IHC interpretation).
How should I quantify GDAP1 across stained sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define regions and cell classes before scoring, since HPA reports different staining levels among Purkinje cells, kidney distal tubules, and other tissues (HPA tissue IHC). Use an H-score for cell intensity and percentage positive, or measure positive cells per mm² when cell density is the question (standard IHC quantification). Normalize cell counts to the analysed tissue area and intensity comparisons to identically processed reference sections and matched development settings (standard IHC practice). Exclude folds, missing tissue, and poorly preserved regions using the same rules for every slide (standard IHC practice). Report compartment and cell type alongside the numeric result rather than pooling distinct structures (UniProt Q8TB36 localisation; HPA tissue IHC).
How can I separate true GDAP1 signal from staining artefacts?
A plausible positive signal is cell-associated and cytoplasmic or membrane-associated in an expected cell population, consistent with GDAP1 localisation and HPA tissue patterns (UniProt Q8TB36 localisation; HPA tissue IHC). Kidney distal tubules and cerebellar Purkinje cells provide reported High-staining comparisons, while HPA reports adipocytes in adipose tissue as Not detected (HPA tissue IHC). Treat isolated nuclear staining, section-edge colour, and staining concentrated in necrotic areas as reasons to investigate artefact (UniProt Q8TB36 localisation; standard IHC practice). Compare no-primary controls to identify endogenous enzyme or detection-related colour, then inspect adjacent intact tissue (standard chromogenic IHC practice). HPA rates tissue evidence Enhanced but describes only Medium consistency with RNA expression, so morphology and controls remain important (HPA tissue IHC).
Boster reagents
Best GDAP1 / Ganglioside-induced differentiation-associated protein 1 IHC Antibodies
A03114 has IHC images from paraffin-embedded rat kidney, rat heart, and mouse kidney (catalog IHC image captions). Human, Mouse, and Rat reactivity is listed (catalog reactivity).
Real IHC data
Anti-GDAP1 Antibody
Cat # A03114
A03114 has IHC images of paraffin-embedded rat kidney, rat heart, and mouse kidney (A03114 IHC image captions). IHC is a listed application, while no IF image is supplied (catalog applications; catalog IF image alts).
Which to pick: Choose A03114 for paraffin-section tissue IHC: its own captions show staining at 1:100 after microwave retrieval in 10 mM PBS, pH 7.2; the fixative is unreported (A03114 IHC image captions). For cross-species planning, A03114 lists Human, Mouse, and Rat reactivity, but its IHC images cover only Mouse and Rat; it has a Rabbit host and no reported clone (catalog reactivity and host; A03114 IHC image captions). No SKU in this payload has a listed IF/ICC application or IF image, so IF/ICC use would require independent validation (catalog applications; catalog IF image alts).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8TB36 (GDAP1_HUMAN, Ganglioside-induced differentiation-associated protein 1).
- Human Protein Atlas. GDAP1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. GDAP1 subcellular location (ICC-IF): Mainly localized to the mitochondria. In addition localized to the cytosol..
- Human Protein Atlas. GDAP1 antibody validation summary (2 antibodies).
- Cell expression of GDAP1 in the nervous system and pathogenesis of Charcot-Marie-Tooth type 4A disease. Journal of cellular and molecular medicine 2008 — PMC2570022.
- Ganglioside-induced differentiation associated protein 1 is a regulator of the mitochondrial network: new implications for Charcot-Marie-Tooth disease. The Journal of cell biology 2005 — PMC2171517.
- Lack of GDAP1 induces neuronal calcium and mitochondrial defects in a knockout mouse model of charcot-marie-tooth neuropathy. PLoS genetics 2015 — PMC4393229.
- The Gdap1 knockout mouse mechanistically links redox control to Charcot-Marie-Tooth disease. Brain : a journal of neurology 2014 — PMC3927703.
- PubMed PMID:10217254 — UniProt-cited evidence.
- PubMed PMID:20685671 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GDAP1 (Ganglioside-induced differentiation-associated protein 1). Expected localisation Cytoplasmic IHC (HPA tissue IHC); mitochondrial outer membrane expected (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A03114 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells - cytoplasm/membrane); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat kidney tissue; fixative not specified (datasheet A03114); 2. Fixation: Image fixative and duration unreported (datasheet A03114); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 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-GDAP1, 1:50-1:200 (datasheet A03114); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope effects are unknown (UniProt); Regulation: No staining regulator reported (UniProt). Evidence: UniProt Q8TB36, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the mitochondria. In addition localized to the…, Datasheet A03114, PMC2570022. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GDAP1/immunohistochemistry-ihc-guide.