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GCC2 / GRIP and coiled-coil domain-containing protein 2 · IHC design guide
Design Immunohistochemistry for GCC2
Plan GCC2 paraffin IHC around its widespread cytoplasmic tissue staining (HPA tissue IHC), while treating trans-Golgi network localization as molecular context (UniProt). The catalog antibody has an IHC dilution range of 1:50–1:200 (datasheet); high-staining glandular cells and listed undetected populations offer comparison points (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GCC2 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining in tissue sections (HPA tissue IHC) | |
| Staining pattern | Widespread cytoplasmic staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Oral mucosa+1 more · see all |
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 and RNA expression show medium consistency (HPA tissue IHC) | |
| Regulation | Low tissue specificity by RNA expression (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms; assess antibody epitope coverage (UniProt) |
Section 1
Recommended GCC2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by two published GCC2 IHC protocols: lung tissue (PMC13145344 methods) and human kidney tissue (PMC11203101 methods).
| Sample | Paraffin-embedded human colon tissue; fixative not specified (datasheet A07107) |
| Fixation | Image fixative and duration unreported (datasheet A07107); 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-GCC2, 1:50-1:200 (datasheet A07107) |
| 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 | GCC2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen retrieval). Both articles report heat retrieval without specifying its conditions (PMC13145344 methods; PMC11203101 methods).
Section 2
What Is the Expected GCC2 Staining Pattern?
GCC2 is a cytoplasmic protein associated with the trans-Golgi network membrane despite having no transmembrane segment (UniProt Q8IWJ2: location and topology). In paraffin-section IHC, expect cytoplasmic staining across many cell types, with especially strong staining in the listed glandular and epithelial cells and cerebellar Purkinje cells (HPA: ubiquitous cytoplasmic expression; listed High cells). HPA rates the tissue IHC evidence Enhanced, while reporting medium consistency with RNA expression (HPA: tissue IHC reliability).
| Discrete cytoplasmic staining, often concentrated near the nucleus, in colon or duodenal glandular cells. | This fits GCC2’s trans-Golgi network association and the High staining reported for these cells (UniProt Q8IWJ2: location; HPA: colon and duodenum glandular cells High). Judge a compact Golgi-like focus in the context of the surrounding cytoplasmic signal; routine chromogenic IHC may not resolve the Golgi as precisely as ICC-IF (standard microscopy practice). |
| Predominantly nuclear staining, with little convincing cytoplasmic signal. | Recheck specificity and scoring because cytoplasmic expression is the tissue IHC profile and the Golgi is the main ICC-IF location (HPA: tissue IHC profile; HPA: ICC-IF main location). Do not automatically dismiss a minor nuclear component: HPA also supports additional nucleoplasmic localization in ICC-IF (HPA: ICC-IF additional location). |
| Strong staining in oral squamous epithelial cells or splenic red-pulp cells. | These cells were reported as Not detected, so strong staining is discordant with the supplied tissue observations (HPA: oral mucosa squamous cells and spleen red-pulp cells Not detected). Consider cross-reactivity or endogenous chromogen-generating activity, then compare controls and staining distribution (standard IHC practice). A negative HPA observation does not establish absolute biological absence. |
| Diffuse color across tissue, extracellular areas, and unrelated cell types. | A widespread haze without a cellular pattern is harder to assign to GCC2 than cytoplasmic staining in the expected cells (HPA: ubiquitous cytoplasmic expression; standard IHC interpretation). Assess background from nonspecific binding or the detection system with an appropriate negative control, and score cellular signal only after that comparison (standard IHC practice). |
| No staining in an adequately preserved colon or duodenum section. | Both contain glandular cells scored High by HPA, so absent signal calls the assay result into question (HPA: colon and duodenum glandular cells High). Confirm that glandular cells are present, the detection controls worked, and the section was processed as intended before calling the specimen GCC2-negative (standard IHC practice). |
💡Expected GCC2 appearanceCall a positive result when glandular or epithelial cells show clear cytoplasmic, preferably Golgi-like staining with strong signal in an HPA High control; broad acellular color or isolated strong staining in HPA Not detected cells warrants investigation (HPA: tissue IHC profile and listed cell levels; UniProt Q8IWJ2: location).
| Tissue and cell selection | High staining is documented in adrenal, appendix, colon, duodenum, endometrium, and epididymis glandular cells, bronchial respiratory epithelium, and cerebellar Purkinje cells (HPA: tissue IHC High cells). Cervical glandular cells, fibroblasts in soft tissue, and adipocytes are reported Low; avoid expecting equal intensity across these cell types (HPA: tissue IHC Low cells). |
| Compartment and topology | GCC2 is associated with the trans-Golgi network membrane and has no transmembrane segment (UniProt Q8IWJ2: location and topology). Interpret a cytoplasmic or Golgi-like pattern accordingly, without treating absence of a cell-surface rim as assay failure (UniProt Q8IWJ2: location; standard IHC interpretation). |
| Antibody validation | HPA reports Enhanced IHC validation for both HPA035849 and HPA035850, and Enhanced reliability for the tissue staining profile (HPA: antibody validation; HPA: tissue IHC reliability). That supports the observed pattern, while HPA’s medium staining–RNA consistency limits how strongly any single weak or negative cell population can be interpreted (HPA: reliability description). |
| IF/ICC: what pattern should corroborate IHC? | A Golgi apparatus signal is the principal ICC-IF finding; an additional nucleoplasmic signal is supported (HPA: ICC-IF main and additional locations). This provides a compartment cross-check for IHC interpretation, while the paraffin-section decision should still be based on its own cellular pattern and controls (HPA: tissue IHC profile; standard IHC practice). |
| Isoforms and antigen retrieval | UniProt lists 2 isoforms, but the supplied record does not map the IHC antibody epitope to either one (UniProt Q8IWJ2: isoforms; supplied antibody record). Antigen retrieval may be optimized against a known-positive section as general paraffin-IHC practice; no GCC2-specific retrieval condition or fixation sensitivity is established here (standard IHC practice; supplied record). |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive-control glandular cells are blank. | The expected cells may be absent from the section, or a staining step may have failed; colon and duodenum glandular cells are reported High (HPA: colon and duodenum). | Check the tissue morphology and run an appropriate detection control; then review retrieval, antibody incubation, and detection steps using the validated assay instructions (standard IHC practice). |
| Only nuclei are strongly stained. | This conflicts with the principal tissue cytoplasmic and ICC-IF Golgi patterns, although additional nucleoplasmic ICC-IF localization is supported (HPA: tissue IHC profile; HPA: ICC-IF locations). | Compare a known-positive section and negative control, and reassess whether any cytoplasmic or Golgi-like signal remains before assigning nuclear staining to GCC2 (standard IHC practice). |
| Color appears throughout the section without cell boundaries. | Nonspecific binding or detection background can obscure the expected cellular cytoplasmic pattern (standard IHC practice; HPA: tissue IHC profile). | Inspect a negative control, review blocking and washing, and adjust detection conditions according to the assay instructions (standard IHC practice). |
| Strong color appears in oral squamous cells or splenic red pulp. | Those populations are reported Not detected; cross-reactivity or endogenous detection activity is possible (HPA: oral mucosa and spleen; standard IHC practice). | Compare no-primary and appropriate detection controls, inspect whether staining follows cell boundaries, and confirm the pattern in an HPA High control (standard IHC practice; HPA: listed High cells). |
| A High control stains, but a test tissue looks weak. | Expression varies by cell type: HPA reports Low staining in cervical glandular cells, soft-tissue fibroblasts, and adipocytes (HPA: tissue IHC Low cells). | Identify the scored cell population and compare equivalent cells at the same detection settings; record weak cellular staining separately from diffuse background (standard IHC practice). |
| The Golgi-like focus is difficult to distinguish in chromogenic IHC. | The trans-Golgi network can be harder to resolve in paraffin sections than in ICC-IF images (UniProt Q8IWJ2: location; standard microscopy practice). | Score the cellular cytoplasmic pattern alongside a known-positive tissue and controls; use the HPA ICC-IF localization as a compartment reference, not as a paraffin-section protocol (HPA: ICC-IF main location; standard IHC practice). |
Experiment evidence
IHC & IF images showing GCC2 staining
A07107 · IHCImmunohistochemistry of paraffin-embedded human colon using GCC2 antibody at dilution of 1:150 .Perform microwave antigen retrieval with 10
A07107 · IHCImmunohistochemistry of paraffin-embedded human stomach using GCC2 antibody at dilution of 1:150 .Perform microwave antigen retrieval with 1
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000135968-GCC2/tissue/cerebral+cortex#imid_9004034
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000135968-GCC2/tissue/colon#imid_8997248
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000135968-GCC2/tissue/liver#imid_8997249
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000135968-GCC2/tissue/kidney#imid_8997258
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035849; green = GCC2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035849; green = GCC2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GCC2 IHC & IF
🧪Run colon first: its glandular cells should stain (HPA: High in colon glandular cells; selected A07107 tissue-IHC caption: human colon). Run oral mucosa as the negative tissue (HPA: Not detected in oral mucosa squamous epithelial cells); on the colon slide, assess nonglandular cells as internal background comparators without assuming they are GCC2-negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Oral mucosa (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GCC2 in A-431, U-251MG, U2OS, SiHa, with annotated localisation: Golgi apparatus (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an immunoglobulin control matched to the primary antibody’s host species and class; use nonimmune IgG for a polyclonal primary. Confirm specificity with a GCC2 knockout sample or peptide block, and quench endogenous peroxidase and inspect any residual chromogenic signal in colon tissue.
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected A07107 tissue-IHC caption does not state a fixative. The caption reports microwave antigen retrieval in 10 mM PBS, pH 7.2, before paraffin-section IHC; it does not establish whether retrieval is required (selected A07107 tissue-IHC caption). Frozen sections cannot be judged easier from these data; IF/ICC can assess the expected Golgi pattern (HPA: enhanced Golgi localization), while colon background should be checked with the no-primary control.
HPA tissue IHC evidence for GCC2
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced GCC2 IHC Tips
Troubleshoot GCC2 staining in paraffin sections by checking retrieval, cellular pattern and controls before assigning a chromogenic score.
How should I retrieve GCC2 in paraffin sections with weak staining?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page HIER rule: cytoplasmic / membrane antigen). Allow sections to cool in buffer, then compare staining with a matched section processed without retrieval so that any gain in signal can be distinguished from increased background (standard IHC practice). If staining remains weak, test microwave retrieval in 10 mM PBS, pH 7.2, the condition shown for catalog antibody A07107 in paraffin-embedded human colon (A07107 tissue-IHC caption). Keep antibody dilution and chromogen development constant across the comparison; the caption used 1:150, but it does not establish that either retrieval condition works equally well in other specimens (A07107 tissue-IHC caption).
Could fixation explain weak or uneven GCC2 staining?
GCC2-specific sensitivity to fixation is unknown because no target-specific fixation condition is supplied here; the catalog image identifies paraffin-embedded human colon but does not state its fixative (A07107 tissue-IHC caption). Record the fixative, fixation duration and section processing for each specimen, and compare sections processed together before changing antibody concentration (standard IHC practice). Excessive or uneven fixation can alter epitope accessibility in IHC generally, so test retrieval on matched sections while keeping detection and chromogen development constant (standard IHC practice). Do not infer a GCC2 fixation preference from its Golgi location, modifications or tissue staining pattern (UniProt Q8IWJ2 subcellular location and modified residues; HPA tissue IHC).
What staining pattern supports GCC2 localisation in tissue sections?
Assess staining within cells for a concentrated perinuclear cytoplasmic pattern consistent with the Golgi, while allowing for additional cytoplasmic signal (UniProt Q8IWJ2 subcellular location; HPA subcellular: Golgi apparatus enhanced). GCC2 associates with the trans-Golgi network membrane but has no transmembrane segment, so a continuous cell-surface outline alone is a poor match for its recorded location (UniProt Q8IWJ2 topology and subcellular location). HPA also reports supported nucleoplasmic localisation in cell imaging; interpret nuclear chromogen cautiously in tissue until an orthogonal control supports it (HPA subcellular). Compare the compartment and cellular distribution with a positive tissue control on the same run, rather than calling all brown cytoplasm specific (standard IHC practice).
How should epitope location affect my GCC2 antibody choice?
Confirm the immunogen or mapped epitope before comparing antibodies, because GCC2 has 2 reported isoforms and a GRIP domain at residues 1609–1659 (UniProt Q8IWJ2 isoforms and domains). An antibody directed to a sequence absent from one isoform may miss that isoform; determine sequence coverage from the antibody documentation rather than assuming both are detected (UniProt Q8IWJ2 isoforms; standard antibody validation practice). Retrieval can change access to a fixed epitope, so compare candidate conditions on adjacent sections with the same detection settings (standard IHC practice). GCC2 has reported phosphorylation and N-terminal acetylation sites; without a mapped epitope, do not assign a staining difference to either modification (UniProt Q8IWJ2 modified residues).
How can I check the GCC2 pattern by multiplex immunofluorescence?
Use a separate IF/ICC validation workflow and pair GCC2 with a marker identifying the expected cell type in the specimen; the colon IHC image alone establishes no IF performance (A07107 tissue-IHC caption; standard IF practice). Choose spectrally separated fluorophores and place the weaker channel away from the tissue’s strongest autofluorescence, using single-stain and no-primary controls to assess bleed-through and background (standard IF practice). Optimise permeabilisation for access to the epitope: GCC2 is cytoplasmic and associated with the trans-Golgi network membrane, with no transmembrane segment, but the antibody’s epitope side is unspecified (UniProt Q8IWJ2 topology and subcellular location). Look for Golgi-enriched signal and evaluate any nuclear signal against controls (HPA subcellular).
How do I reduce diffuse brown background without losing GCC2 signal?
First compare the stained section with a no-primary control and inspect whether diffuse chromogen also appears where cellular GCC2 signal is absent (standard IHC practice). Block endogenous peroxidase before peroxidase-based detection, then review blocking, washes and chromogen development if the control is brown (standard chromogenic IHC practice). Titrate the catalog antibody around the documented 1:150 colon image condition while holding retrieval and detection constant; that image does not establish an optimum for every specimen (A07107 tissue-IHC caption). GCC2 is reported as ubiquitously cytoplasmic in tissue, so widespread staining alone is insufficient evidence of background; judge its cellular pattern and control staining together (HPA tissue IHC).
How should I quantify GCC2 chromogenic staining across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then record the percentage of positive cells and staining intensity with identical thresholds across slides (standard IHC scoring practice). An H-score can combine percentage at each intensity as 0 × % negative + 1 × % weak + 2 × % moderate + 3 × % strong, giving a range of 0–300 (standard IHC scoring practice). Normalise positive-cell counts or stained area to the number of eligible cells or tissue area in mm², and exclude folds, edges and necrotic regions consistently (standard image-analysis practice). Report Golgi-enriched cytoplasmic and nuclear scores separately because GCC2’s main Golgi localisation and additional nucleoplasmic signal have different evidence levels (HPA subcellular).
When is GCC2 staining convincing rather than an artefact?
A convincing result combines intracellular Golgi-enriched staining with an appropriate cellular distribution and clean negative controls (UniProt Q8IWJ2 subcellular location; HPA subcellular; standard IHC practice). High staining in colon glandular cells can support a positive control, whereas oral mucosa squamous epithelial cells were reported as not detected; these observations do not guarantee the same result with every antibody or preparation (HPA tissue IHC). Treat isolated surface outlines, section-edge gradients and staining confined to necrotic areas as suspect until replicated away from those regions (UniProt Q8IWJ2 topology; standard IHC practice). If brown signal persists without primary antibody, investigate endogenous enzyme activity and detection background before assigning GCC2 positivity (standard chromogenic IHC practice).
Boster reagents
Best GCC2 / GRIP and coiled-coil domain-containing protein 2 IHC Antibodies
A07107 has IHC images from paraffin-embedded human colon and stomach (catalog image captions). It is listed as reactive with human and mouse samples (catalog reactivity).
Real IHC data
Anti-GCC2 Antibody
Cat # A07107
A07107 will render with a human colon IHC image; the catalog also provides a human stomach IHC image, both using paraffin sections (catalog image captions). Its listed applications are IHC and WB, and its listed reactivity is human and mouse (catalog applications; catalog reactivity).
Which to pick: Choose A07107 for tissue IHC on paraffin sections; its images show human colon and stomach, while the fixative is unreported (catalog image captions). For mouse tissue, A07107 has listed reactivity but no mouse IHC image in the payload (catalog reactivity; catalog image captions). No IF/ICC application or image is listed for A07107, so this payload does not support an IF/ICC pick (catalog applications; catalog image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8IWJ2 (GCC2_HUMAN, GRIP and coiled-coil domain-containing protein 2).
- Human Protein Atlas. GCC2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. GCC2 subcellular location (ICC-IF): Mainly localized to the Golgi apparatus. In addition localized to the nucleoplasm..
- Human Protein Atlas. GCC2 antibody validation summary (2 antibodies).
- GCC2 in Small Extracellular Vesicles as a Diagnostic and Prognostic Biomarker of Early-Stage Lung Adenocarcinoma. Journal of extracellular vesicles 2026 — PMC13145344.
- LIM Zinc Finger Domain Containing 1 Risk Genotype of Recipient Is Associated with Renal Tubular Inflammation in Kidney Transplantation. Genes 2024 — PMC11203101.
- PubMed PMID:9205841 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GCC2 (GRIP and coiled-coil domain-containing protein 2). Expected localisation Cytoplasmic staining in tissue sections (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A07107 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Oral mucosa. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon tissue; fixative not specified (datasheet A07107); 2. Fixation: Image fixative and duration unreported (datasheet A07107); 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-GCC2, 1:50-1:200 (datasheet A07107); 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 and RNA expression show medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; assess antibody epitope coverage (UniProt); Regulation: Low tissue specificity by RNA expression (HPA tissue RNA). Evidence: UniProt Q8IWJ2, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the Golgi apparatus. In addition localized to…, Datasheet A07107, PMC13145344, PMC11203101. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GCC2/immunohistochemistry-ihc-guide.