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GGCX / Vitamin K-dependent gamma-carboxylase · IHC design guide
Design Immunohistochemistry for GGCX
Plan GGCX paraffin-section IHC using hepatocyte staining in liver as a positive reference (HPA tissue IHC). The guide covers the catalog antibody’s chromogenic workflow (datasheet A02643-1) and how to interpret cytoplasmic staining in light of GGCX’s ER membrane location (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GGCX 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 tissue signal; ER membrane protein (HPA tissue IHC; UniProt) | |
| Staining pattern | Hepatocytes show the strongest cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02643-1) | |
| Positive control | Liver+4 more · see all | |
| Negative control | Fallopian tube |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Endogenous biotin may raise liver background with SABC (standard IHC practice; datasheet A02643-1) | |
| Regulation | Liver-enriched expression (HPA tissue RNA; HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; epitope mapping is needed to assess coverage (UniProt) |
Section 1
Recommended GGCX IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by one published GGCX liver IHC protocol (PMC12084421).
| Sample | Paraffin-embedded human hepatic cancer tissue; fixative not specified (datasheet A02643-1) |
| Fixation | Image fixative and duration unreported (datasheet A02643-1); 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 A02643-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02643-1) |
| Primary antibody | Rabbit anti-GGCX, 2-5 μg/ml (datasheet A02643-1) |
| Primary incubation | Overnight at 4 °C (datasheet A02643-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02643-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GGCX-positive staining in hepatocytes of liver (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several different tissue types, most abundant in hepatocytes. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A02643-1); use citrate microwave retrieval when reproducing the liver study (PMC12084421).
Section 2
What Is the Expected GGCX Staining Pattern?
GGCX is an endoplasmic reticulum membrane protein with nine transmembrane segments (UniProt P38435 topology). In paraffin-section IHC, expect cytoplasmic staining, strongest in liver hepatocytes (HPA tissue IHC: High in hepatocytes). HPA also reports Medium staining in several other cell types. Its tissue IHC reliability is Enhanced, with medium consistency between antibody staining and RNA expression (HPA tissue IHC: reliability summary).
| Hepatocytes show prominent cytoplasmic chromogen, while nuclei remain defined by the counterstain. | This is the clearest expected positive pattern: hepatocytes are High and the reported distribution is cytoplasmic (HPA tissue IHC). The ER membrane location explains why GGCX signal is assessed in the cytoplasmic compartment (UniProt P38435 subcellular location). |
| Adrenal or breast glandular cells, bronchial respiratory epithelial cells, or cerebellar Purkinje cells show moderate cytoplasmic staining. | These are plausible positive results; each listed cell population is Medium in the HPA tissue IHC record (HPA tissue IHC). Judge the identified cell population and compartment, rather than requiring every tissue to match hepatocyte intensity. |
| Nuclear staining dominates, or a crisp cell-surface rim replaces the expected cytoplasmic pattern. | Treat this as a compartment mismatch and investigate artefact or nonspecific detection: HPA describes cytoplasmic tissue staining, and UniProt places GGCX in the ER membrane (HPA tissue IHC; UniProt P38435 subcellular location). The slide alone cannot identify the cause. |
| Strong chromogen appears in cells outside the reported pattern, or broadly across unrelated structures. | Consider cross-reactivity or endogenous detection activity and check controls (standard IHC practice). HPA reports specific cell populations, including Not detected in fallopian-tube glandular cells; that reference is a comparison, not proof that every unlisted cell is negative (HPA tissue IHC). |
| Color covers tissue and empty spaces diffusely, or hepatocytes show no discernible signal. | Diffuse background limits localization and warrants detection and wash controls (standard IHC practice). Absent signal in hepatocytes conflicts with their High HPA staining level; first assess section quality, controls, and the IHC workflow before interpreting GGCX as absent (HPA tissue IHC; standard IHC practice). |
💡Expected GGCX appearanceCall positive IHC when hepatocytes show strong cytoplasmic staining consistent with ER localization; dominant nuclear, edge-only, or structure-wide color is suspect (HPA tissue IHC: High hepatocytes, cytoplasmic profile; UniProt P38435 subcellular location; standard IHC practice).
| Subcellular topology | GGCX has 9 transmembrane segments and is assigned to the ER membrane (UniProt P38435 topology). Interpret the HPA cytoplasmic IHC profile in that context; paraffin-section chromogen does not itself resolve individual ER membranes (HPA tissue IHC; standard IHC practice). |
| Anatomic comparison | Hepatocytes are High, several listed epithelial or glandular populations and Purkinje cells are Medium, and fallopian-tube glandular cells are Not detected (HPA tissue IHC). Compare matched cell types rather than whole-section darkness (standard IHC practice). |
| Validation and its limit | The listed antibody HPA018284 has Enhanced IHC validation, while tissue IHC has medium antibody-to-RNA consistency (HPA antibodies; HPA tissue IHC reliability). These assessments support a reference pattern but cannot certify each new section or detection run. |
| Molecular annotations | UniProt lists 2 isoforms and four glycosylation sites at 459, 550, 570, and 605 (UniProt P38435). Neither record supplies the IHC antibody epitope, so these annotations cannot predict isoform selectivity, retrieval requirements, or a staining difference. |
| IF/ICC evidence | HPA's ICC-IF summary says Membrane, but gives no main location or cell-line images (HPA subcellular). Use the ER assignment as biological context, without treating that summary as an imaged cell-line staining pattern (UniProt P38435 subcellular location). |
| Situation | Likely cause | Next action |
|---|---|---|
| Hepatocytes are negative despite an otherwise readable section. | This disagrees with their High reference signal (HPA tissue IHC); one slide cannot distinguish technical failure from specimen variation. | Confirm the positive control and counterstained morphology, then review the IHC antibody dilution, retrieval, detection reagents, and run records (standard IHC practice). No GGCX-specific retrieval setting is supplied. |
| The whole section has weak, even brown haze. | Diffuse color can reflect background from detection chemistry, blocking, or washing (standard IHC practice); HPA's cell-specific profile cannot validate a uniform haze (HPA tissue IHC). | Compare a no-primary control, examine background outside cells, and review blocking, reagent handling, and washes (standard IHC practice). Score GGCX only where cellular localization remains interpretable. |
| Unexpected cells stain as strongly as hepatocytes. | Cross-reactivity or endogenous detection activity is possible (standard IHC practice). HPA documents High hepatocytes and Medium or absent signal in specified other populations, not a complete map of every cell (HPA tissue IHC). | Identify the stained cell type on the counterstained section; compare the HPA reference and no-primary control before assigning GGCX positivity (HPA tissue IHC; standard IHC practice). |
| Nuclei or tissue edges dominate the signal. | That distribution conflicts with cytoplasmic IHC staining and ER membrane localization (HPA tissue IHC; UniProt P38435 subcellular location). Artefact is possible, but its mechanism is unresolved. | Inspect morphology and the no-primary control, then review section handling, chromogen development, and counterstain balance (standard IHC practice). Do not score the misplaced signal as the expected GGCX pattern. |
| A fallopian-tube glandular sample appears strongly positive. | HPA reports Not detected for that cell population (HPA tissue IHC). A discrepancy may reflect cell identification, background, or a real sample difference; the supplied record cannot decide which. | Verify glandular-cell identity and compare the staining with a hepatocyte positive control and no-primary control (HPA tissue IHC; standard IHC practice). Report the discrepancy rather than forcing a negative score. |
| What should an IF/ICC result look like? | HPA summarizes the subcellular signal as Membrane but supplies no main location or cell-line images (HPA subcellular); UniProt assigns GGCX to the ER membrane (UniProt P38435 subcellular location). | Interpret a cellular membrane-associated signal cautiously against the ER assignment and appropriate controls (UniProt P38435; standard IF practice). The supplied HPA record does not establish a specific imaged IF/ICC pattern. |
Experiment evidence
IHC & IF images showing GGCX staining
A02643-1 · IHCIHC analysis of GGCX using anti-GGCX antibody (A02643-1). GGCX was detected in a paraffin-embedded section of human hepatic cancer tissue. H
A02643-1 · IFIF analysis of GGCX using anti-GGCX antibody (A02643-1). GGCX was detected in an immunocytochemical section of CACO-2 cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000115486-GGCX/tissue/cerebral+cortex#imid_4907796
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000115486-GGCX/tissue/colon#imid_4907730
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000115486-GGCX/tissue/liver#imid_4907739
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000115486-GGCX/tissue/kidney#imid_4907732
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GGCX IHC & IF
🧪Run liver first: hepatocytes should stain (HPA: High in hepatocytes). Use fallopian tube glandular cells as the tissue negative (HPA: Not detected); on the liver slide, morphologically identified cells without signal provide an internal background reference, but should not be assumed to lack GGCX (standard IHC practice).
Positive control tissue: Liver (Hepatocytes, HPA High)
Negative control tissue: Fallopian tube (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for GGCX; derive a cell-line control from the positive tissue's cell type (Hepatocytes) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit isotype controls, plus GGCX knockout tissue or a validated immunizing-peptide block as a biological specificity control (caption: rabbit anti-GGCX; standard IHC practice). In liver, check endogenous peroxidase and biotin background before interpreting the biotin-based DAB signal (caption: biotinylated secondary, streptavidin–biotin complex and DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A02643-1 paraffin-section caption does not state a fixative (selected-SKU caption). That caption uses heat retrieval in EDTA at pH 8.0, but does not establish whether GGCX staining depends on retrieval (selected-SKU caption). The supplied evidence does not establish that frozen sections or IF are easier; for liver IHC, assess endogenous peroxidase and biotin background when using the caption’s detection method (HPA: High in hepatocytes; selected-SKU caption; standard IHC practice).
HPA tissue IHC evidence for GGCX
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 |
|---|---|---|---|---|
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GGCX IHC Tips
Troubleshoot GGCX chromogenic IHC in paraffin sections using the catalog antibody’s documented workflow, ER localisation, and tissue expression profile.
How should I retrieve GGCX in paraffin sections when staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for the catalog antibody (datasheet A02643-1). The documented tissue image then used 2 μg/ml primary antibody overnight at 4°C, so reproduce those conditions before changing retrieval (IHC caption A02643-1). If staining remains weak, compare retrieval conditions on adjacent sections while keeping antibody concentration, detection, and development constant (standard IHC practice). Assess signal in viable hepatocytes, where expression is high, and check that stronger retrieval has not increased diffuse background or damaged tissue morphology (HPA: High in hepatocytes; standard IHC practice).
Can the published staining pattern tell me which fixation preserves GGCX best?
No target-specific fixation sensitivity is established here: the catalog image describes a paraffin section but does not state its fixative (IHC caption A02643-1). Record the fixative, fixation duration, and tissue processing for each specimen before comparing staining intensity across cases (standard IHC practice). When optimising a new specimen series, hold the documented EDTA pH 8.0 retrieval and 2 μg/ml primary concentration constant while assessing fixation conditions on matched material (datasheet A02643-1; IHC caption A02643-1). Judge changes against preserved morphology and staining in viable hepatocytes; neither their HPA expression pattern nor GGCX topology establishes a preferred fixative (HPA: High in hepatocytes; UniProt P38435 topology).
Should GGCX staining look cytoplasmic or sharply restricted to the ER?
GGCX is an endoplasmic reticulum membrane protein with 9 annotated transmembrane segments, so intracellular staining is expected (UniProt P38435 subcellular location and topology). In tissue IHC, the reported pattern is cytoplasmic across several tissues and most abundant in hepatocytes; chromogenic resolution need not delineate individual ER membranes (HPA: tissue IHC profile; standard IHC practice). Compare viable hepatocytes with neighbouring cell populations on the same section while preserving cellular boundaries during interpretation (HPA: High in hepatocytes; standard IHC practice). Predominantly nuclear, extracellular, or uniform acellular deposits warrant review of counterstain, morphology, and detection controls before being called GGCX localisation (UniProt P38435 subcellular location; standard IHC practice).
How could epitope position change GGCX staining in paraffin tissue?
The record lists 2 GGCX isoforms, but the supplied evidence does not identify this antibody’s epitope or establish isoform-specific recognition (UniProt P38435 isoforms; IHC caption A02643-1). GGCX has 9 membrane spans and a lumenal region at residues 378–758, making epitope location relevant when evaluating access after processing and retrieval (UniProt P38435 topology). Four annotated glycosylation sites lie at residues 459, 550, 570, and 605; their effect on this antibody’s staining is unknown (UniProt P38435 glycosylation). Request epitope and isoform-reactivity information before interpreting a negative section as absence of every GGCX isoform, and compare it with documented positive tissue (UniProt P38435 isoforms; HPA: High in hepatocytes; standard IHC practice).
How should I adapt GGCX localisation checks to multiplex immunofluorescence?
For an IF experiment, pair GGCX with a validated hepatocyte marker when examining liver, since hepatocytes show high tissue expression (HPA: High in hepatocytes). Choose spectrally separated fluorophores and favour a far-red channel for the weaker signal when tissue autofluorescence interferes; include single-label and unstained controls (standard IF practice). Confirm the antibody’s epitope before tuning permeabilisation: access to a lumenal epitope within residues 378–758 may require membrane permeabilisation, whereas a cytoplasmic epitope presents a different access problem (UniProt P38435 topology; standard IF practice). Optimise fixation and detergent conditions for IF directly, because the supplied catalog evidence documents chromogenic tissue IHC and HPA lists no ICC/IF images (IHC caption A02643-1; HPA: subcellular record).
What should I check when DAB staining spreads beyond hepatocytes?
The documented assay used 10% goat serum, a biotinylated secondary, streptavidin–biotin detection, and DAB (IHC caption A02643-1). First compare sections without primary antibody and evaluate endogenous peroxidase blocking, since either detection chemistry or enzyme activity can produce misleading chromogen (standard IHC practice). If signal persists with the biotin-based system, assess endogenous biotin with an appropriate detection control before changing the primary antibody concentration (IHC caption A02643-1; standard IHC practice). Keep the documented 2 μg/ml primary condition as a reference and judge any blocking adjustment against both background and hepatocyte staining on matched sections (IHC caption A02643-1; HPA: High in hepatocytes; standard IHC practice).
How should I score GGCX staining across liver specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define viable hepatocytes as the primary scoring population and record the percentage positive together with cytoplasmic intensity (HPA: High in hepatocytes; HPA: tissue IHC profile). An H-score combines the percentage of cells at each intensity grade into a 0–300 measure; use one scoring rule, threshold, and image-acquisition setup across cases (standard IHC practice). Normalise positive-cell counts to the number of viable hepatocytes assessed, or report positive-cell density per mm² of viable tissue when cell counts are impractical (standard IHC practice). Exclude necrosis and section edges consistently, and analyse adjacent non-hepatocyte populations separately rather than pooling them into a whole-section value (standard IHC practice; HPA: High in hepatocytes).
How do I distinguish convincing GGCX staining from tissue or detection artefacts?
A convincing signal should be intracellular and compatible with ER-associated protein, with strong hepatocyte staining providing a tissue benchmark (UniProt P38435 subcellular location; HPA: High in hepatocytes). Examine whether apparent positivity instead follows section edges, necrotic areas, or extracellular deposits rather than intact cell boundaries (standard IHC practice). Check a no-primary control and the peroxidase and biotin-related detection controls when brown signal appears broadly across cell types, because the documented assay uses streptavidin–biotin and DAB (IHC caption A02643-1; standard IHC practice). Treat discordant staining cautiously: HPA reports medium agreement between antibody staining and RNA data, while its tissue profile supports a cytoplasmic pattern rather than a nuclear one (HPA: Enhanced reliability description and tissue IHC profile).
Boster reagents
Best GGCX / Vitamin K-dependent gamma-carboxylase IHC Antibodies
A02643-1 has IHC data from a human hepatic cancer paraffin section and IF data from CACO-2 cells (catalog image captions). Its listed reactivity is Human and Mouse (catalog).
Real IHC data
Anti-GGCX Antibody ®
Cat # A02643-1
A02643-1 will render with an IHC figure from a human hepatic cancer paraffin section (IHC image caption). The same SKU is listed for IHC and IF/ICC, with IF data from CACO-2 cells (catalog applications; IF image caption).
Which to pick: Choose A02643-1 for paraffin-section IHC based on its own human hepatic cancer figure; the fixative is unreported (IHC image caption). Choose A02643-1 for IF/ICC based on its CACO-2 figure (IF image caption). It lists Mouse reactivity, but the supplied IHC and IF figures do not establish mouse-sample performance (catalog reactivity; image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P38435 (VKGC_HUMAN, Vitamin K-dependent gamma-carboxylase).
- Human Protein Atlas. GGCX tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. GGCX subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. GGCX antibody validation summary (1 antibodies).
- Identification of liver proteins as biomarker for postmortem diagnosis of heat stroke through proteomics. Scientific reports 2025 — PMC12084421.
- PubMed PMID:1749935 — UniProt-cited evidence.
- PubMed PMID:9166845 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GGCX (Vitamin K-dependent gamma-carboxylase). Expected localisation Cytoplasmic tissue signal; ER membrane protein (HPA tissue IHC; UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02643-1). Validated IHC antibody: A02643-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Liver (Hepatocytes); negative Fallopian tube. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human hepatic cancer tissue; fixative not specified (datasheet A02643-1); 2. Fixation: Image fixative and duration unreported (datasheet A02643-1); 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 A02643-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02643-1); 8. Primary antibody: Rabbit anti-GGCX, 2-5 μg/ml (datasheet A02643-1); 9. Primary incubation: Overnight at 4 °C (datasheet A02643-1). Watch for: Fixation: Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Endogenous biotin may raise liver background with SABC (standard IHC practice; datasheet A02643-1); Isoform / epitope: 2 isoforms; epitope mapping is needed to assess coverage (UniProt); Regulation: Liver-enriched expression (HPA tissue RNA; HPA tissue IHC). Evidence: UniProt P38435, HPA tissue IHC · Enhanced, HPA ICC-IF · Membrane, Datasheet A02643-1, PMC12084421. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GGCX/immunohistochemistry-ihc-guide.