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GGCX / Vitamin K-dependent gamma-carboxylase · IHC design guide
Design Immunohistochemistry for GGCX
Plan GGCX IHC in paraffin sections using hepatocytes as a strong staining reference (HPA tissue IHC). Compare the observed cytoplasmic pattern with its annotated ER membrane localization (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 staining (HPA tissue IHC); ER membrane localization (UniProt). | |
| Staining pattern | Strongest cytoplasmic staining in hepatocytes (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 fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining has medium RNA concordance (HPA tissue IHC). | |
| Regulation | Regulatory changes are unreported (UniProt). | |
| Isoform / epitope | 2 isoforms; check whether the epitope covers both (UniProt). |
Section 1
Recommended GGCX IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet A02643-1). One published GGCX IHC protocol uses citrate retrieval in liver sections (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 heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A02643-1); try microwave citrate retrieval when following the published liver protocol (PMC12084421).
Section 2
What Is the Expected GGCX Staining Pattern?
GGCX is an endoplasmic reticulum membrane protein with 9 transmembrane segments (UniProt P38435 topology). In paraffin-section IHC, expect cytoplasmic staining, strongest in hepatocytes (HPA: tissue IHC). HPA rates its tissue staining reliability as Enhanced, while noting medium consistency between antibody staining and RNA expression (HPA: tissue IHC).
| Strong hepatocyte staining with a cytoplasmic pattern (HPA: High in hepatocytes; cytoplasmic profile). | This best matches the observed tissue pattern and the expected cellular compartment (HPA: tissue IHC; UniProt P38435: ER membrane). Assess the pattern within hepatocytes, rather than treating any brown deposit in liver as specific staining (general IHC practice). |
| Predominantly nuclear staining, or staining confined to the cell surface (UniProt P38435: ER membrane). | That distribution does not fit the annotated ER membrane location or HPA's cytoplasmic tissue profile (UniProt P38435; HPA: tissue IHC). Review morphology and controls before interpreting it as GGCX; misplaced chromogen or nonspecific antibody binding are possible explanations (general IHC practice). |
| Strong glandular-cell staining in fallopian tube, while hepatocytes stain weakly (HPA: fallopian tube Not detected; hepatocytes High). | The contrast runs against HPA's reported cell patterns (HPA: tissue IHC). Consider cross-reactivity or endogenous detection activity, and compare with an appropriate negative detection control (general IHC practice). HPA's negative observation is a comparator, not proof that every specimen must be blank. |
| Broad, diffuse color outside recognizable cell boundaries (general IHC practice). | Extracellular haze cannot establish the cytoplasmic cell-associated pattern reported by HPA (HPA: tissue IHC). Check whether a negative detection control shows similar color; inadequate blocking, residual detection activity, or excess detection reagent can produce background (general IHC practice). |
| No visible hepatocyte signal in an otherwise interpretable liver section (HPA: High in hepatocytes). | This conflicts with the strongest supplied positive tissue observation, but alone does not establish biological absence (HPA: tissue IHC). Check section integrity, antibody application, retrieval conditions, and detection controls as general IHC workflow checks; no GGCX-specific fixation sensitivity is supplied. |
💡Expected GGCX appearanceA convincing positive is high cytoplasmic hepatocyte staining (HPA: tissue IHC), consistent with ER membrane localization (UniProt P38435); dominant nuclear, extracellular, or unexpected negative-control color warrants review (general IHC practice).
| Tissue and cell context (HPA: tissue IHC). | Hepatocytes are High; several listed glandular or epithelial populations are Medium, and fallopian-tube glandular cells are Not detected (HPA: tissue IHC). Compare the named cell population and intensity, not just the tissue label. |
| Membrane topology and epitope location (UniProt P38435 topology). | GGCX has 9 transmembrane segments and both cytoplasmic and lumenal regions (UniProt P38435 topology). Epitope location could affect accessibility under a given assay, but no antibody epitope or target-specific retrieval response is supplied. |
| Isoforms and processing (UniProt P38435). | UniProt lists 2 isoforms and a chain spanning residues 2–758, with no signal peptide or propeptide annotated (UniProt P38435). These facts do not establish isoform-specific staining or a shed extracellular staining pattern. |
| Antibody evidence (HPA: HPA018284 IHC Enhanced). | HPA lists one rabbit polyclonal antibody, HPA018284, with Enhanced IHC validation (HPA: antibody record). HPA also describes medium staining–RNA consistency; interpret an unfamiliar pattern against tissue controls rather than assuming equivalent performance in every specimen (HPA: tissue IHC; general IHC practice). |
| IF/ICC: what localization can this record support? | ER membrane localization is annotated by UniProt (UniProt P38435); HPA summarizes subcellular staining as Membrane but supplies no main location or ICC-IF cell-line images (HPA: subcellular record). An IF/ICC pattern cannot be independently confirmed from those images. |
| Situation | Likely cause | Next action |
|---|---|---|
| Hepatocytes are blank (HPA: hepatocytes High). | A failed assay step is possible, but the supplied sources identify no GGCX-specific fixation or retrieval sensitivity (general IHC practice; source scope). | Check tissue morphology, antibody application, detection reagents, and an assay-positive control; review retrieval as a general IHC variable without claiming a GGCX-specific optimum (general IHC practice). |
| All cells and surrounding tissue show similar color (general IHC practice). | Diffuse background or endogenous detection activity may obscure cell-associated staining (general IHC practice). | Inspect a negative detection control, review blocking and washing, and confirm that color follows cell boundaries before scoring GGCX (general IHC practice; HPA: cytoplasmic tissue profile). |
| Nuclei dominate the signal (UniProt P38435: ER membrane). | The compartment conflicts with the UniProt location and HPA cytoplasmic profile; nonspecific staining or interpretation error is possible (UniProt P38435; HPA: tissue IHC; general IHC practice). | Compare the counterstain and negative control, then assess whether any separate cytoplasmic signal is present in hepatocytes (general IHC practice; HPA: hepatocytes High). |
| Fallopian-tube glandular cells appear strongly positive (HPA: Not detected). | The result disagrees with the supplied HPA observation; cross-reactivity or endogenous detection activity is possible (HPA: tissue IHC; general IHC practice). | Verify the cell identity and compare negative detection controls and hepatocyte staining before assigning the signal to GGCX (general IHC practice; HPA: hepatocytes High). |
| Medium-level tissue staining is scored as absent (HPA: listed Medium populations). | Using the hepatocyte High pattern as the only positive threshold can miss weaker reported populations (HPA: tissue IHC). | Score the specified cell type and intensity against background and controls; HPA reports Medium staining in the listed adrenal, breast, bronchial, cerebellar, duodenal, endometrial, and esophageal populations (HPA: tissue IHC). |
| A new specimen shows a pattern unlike HPA's examples (HPA: tissue IHC). | HPA reports medium consistency between antibody staining and RNA expression, so the supplied evidence does not settle an unfamiliar result (HPA: reliability description). | Document the cell compartment, intensity, and controls; compare with the reported hepatocyte positive and fallopian-tube glandular-cell negative patterns before drawing a target-specific conclusion (HPA: tissue IHC; general IHC practice). |
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 and score hepatocytes for GGCX staining (HPA: High in hepatocytes). Run fallopian tube glandular cells as the tissue negative (HPA: Not detected); on the liver slide, treat cells outside the scored hepatocytes as background comparators, not validated internal negatives (HPA: High in hepatocytes).
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 a no-primary, secondary-only control, a matched rabbit isotype control, and a GGCX knockout or antigen-peptide blocking control (caption: rabbit primary antibody; standard IHC practice). In liver, check endogenous peroxidase and biotin background before interpreting chromogenic staining (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact A02643-1 paraffin-section caption does not state a fixative (selected-SKU caption). Heat retrieval in EDTA at pH 8.0 was used for the pictured IHC section, but retrieval dependency is unreported (selected-SKU caption). The supplied evidence does not establish whether frozen sections or IF are easier; liver endogenous biotin can complicate the caption’s biotin-based detection scheme (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
Use the documented paraffin-section method as a starting point, then assess GGCX staining against its expected cellular distribution and appropriate controls (caption A02643-1; UniProt P38435 topology).
How should I troubleshoot weak GGCX staining after antigen retrieval?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A02643-1). The selected tissue image used this retrieval before overnight incubation with 2 μg/ml catalog antibody at 4°C (caption A02643-1). If staining is weak, compare heating durations on adjacent sections while holding antibody concentration and DAB development constant (standard IHC practice). Record tissue integrity and hepatocyte cytoplasmic signal as you assess the result (HPA: High in hepatocytes; standard IHC practice). Compare another buffer only as a fallback, and include a no-primary control to assess retrieval-associated background (standard IHC practice).
What fixation conditions preserve interpretable GGCX staining in paraffin sections?
Target-specific fixation sensitivity for GGCX is unknown because the selected paraffin-section caption does not state a fixative (caption A02643-1). Record the actual fixative, delay before fixation, fixation duration, and section age for each block (standard IHC practice). To assess fixation effects, compare similarly processed samples while varying one fixation condition at a time (standard IHC practice). Apply the documented EDTA pH 8.0 retrieval and 2 μg/ml antibody conditions consistently across that comparison (datasheet A02643-1; standard IHC practice). Assess morphology alongside staining, and avoid calling reduced DAB intensity protein loss when processing differs (standard IHC practice).
Where should convincing GGCX staining appear in a paraffin section?
Expect a cytoplasmic staining pattern, especially in hepatocytes, while interpreting GGCX as an endoplasmic-reticulum membrane protein (HPA: High in hepatocytes and cytoplasmic expression; UniProt P38435 subcellular location). Its 9 annotated transmembrane segments support an intracellular membrane location, but chromogenic IHC need not resolve individual ER membranes (UniProt P38435 topology; standard IHC practice). Compare signal within morphologically intact cells and across similarly processed sections (standard IHC practice). Markedly nuclear or extracellular DAB staining warrants scrutiny with a no-primary control (UniProt P38435 subcellular location; standard IHC practice). Preserve the distinction between a cytoplasmic light-microscopy pattern and molecular proof of ER localization (standard IHC practice).
How do topology and isoforms affect interpretation of this antibody's staining?
GGCX has 2 annotated isoforms and 9 transmembrane segments, so epitope placement matters when interpreting staining (UniProt P38435 isoforms and topology). The record places residues 378–758 on the luminal side and 306–364 on the cytoplasmic side (UniProt P38435 topology). It also lists glycosylation sites at 459, 550, 570, and 605, all within that annotated luminal region (UniProt P38435 topology and glycosylation). The supplied antibody caption does not identify its epitope or establish which isoforms it detects (caption A02643-1). Seek epitope and isoform coverage documentation before assigning differences in DAB intensity to isoform expression or glycosylation (standard IHC practice).
How should I adapt the GGCX question to multiplex immunofluorescence?
Treat IF/ICC performance as unestablished by the supplied paraffin-section IHC caption, and validate the antibody separately for fluorescence imaging (caption A02643-1; standard IF practice). Multiplex GGCX with a hepatocyte marker when testing liver, because hepatocytes show high tissue staining (HPA: High in hepatocytes; standard IF practice). Select fluorophores after examining unstained tissue for autofluorescence, and reserve spectrally separated channels for the two targets (standard IF practice). Choose permeabilisation with the mapped epitope in mind: the record includes both cytoplasmic and luminal regions across 9 transmembrane segments (UniProt P38435 topology; standard IF practice). Compare permeabilised and untreated controls before inferring absent GGCX from weak fluorescence (standard IF practice).
How can I separate GGCX signal from chromogenic background?
The selected method used 10% goat serum blocking, a biotinylated secondary antibody, an avidin–biotin detection complex, and DAB (caption A02643-1). Include a no-primary section and a peroxidase block to assess signal unrelated to primary-antibody binding (standard IHC practice). If background persists, compare detection with and without the biotin-based amplification step to assess that component (caption A02643-1; standard IHC practice). Keep DAB development time and section handling consistent while changing one blocking or detection condition at a time (standard IHC practice). Judge improvement by retained hepatocyte cytoplasmic signal together with reduced staining outside viable cells (HPA: High in hepatocytes; standard IHC practice).
How should I score GGCX staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population before comparing slides; hepatocytes are a relevant reference because their GGCX staining is high in the tissue atlas (HPA: High in hepatocytes; standard IHC practice). For cellular expression, record the percentage of positive viable cells and an intensity-based H-score from 0–300 using a consistent scoring scheme (standard IHC practice). Normalize counts to the number of eligible viable cells, or report positive-cell density per mm² of viable tissue (standard IHC practice). Keep retrieval, antibody exposure, and DAB development matched across sections (datasheet A02643-1; standard IHC practice). Report cell type, compartment, excluded areas, and scoring rules alongside the numerical result (standard IHC practice).
What findings distinguish genuine GGCX staining from artefact?
Give greatest weight to reproducible cytoplasmic staining in viable hepatocytes, consistent with the tissue atlas and GGCX's ER membrane location (HPA: High in hepatocytes and cytoplasmic expression; UniProt P38435 subcellular location). Treat isolated nuclear or extracellular signal as suspect until controls and morphology support it (UniProt P38435 subcellular location; standard IHC practice). Inspect section edges and necrotic areas separately because staining there can reflect processing artefact (standard IHC practice). Use a no-primary control and peroxidase block to investigate DAB signal unrelated to the primary antibody (standard IHC practice). Interpret differences cautiously: the atlas assigns its tissue staining “Enhanced” reliability while describing only medium agreement with RNA expression (HPA: reliability description).
Boster reagents
Best GGCX / Vitamin K-dependent gamma-carboxylase IHC Antibodies
The IHC-validated antibody has real paraffin-section IHC data from human hepatic cancer tissue and IF/ICC data from CACO-2 cells; human and mouse reactivity is listed (catalog; IHC and IF captions).
Real IHC data
Anti-GGCX Antibody ®
Cat # A02643-1
A02643-1 will render with IHC data from a paraffin-embedded human hepatic cancer section (IHC caption). The same SKU is listed for IF/ICC and has an IF image from CACO-2 cells; the catalog lists human and mouse reactivity (IF caption; catalog).
Which to pick: Choose A02643-1 for paraffin-section IHC: its image caption reports EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml; the fixative is unreported (IHC caption). Choose A02643-1 for IF/ICC as well: its CACO-2 image uses 5 μg/ml primary antibody (IF caption). For cross-species planning, A02643-1 lists human and mouse reactivity, while the supplied IHC and IF images show human samples only (catalog; IHC and IF 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 staining (HPA tissue IHC); ER membrane localization (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 fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Antibody staining has medium RNA concordance (HPA tissue IHC).; Isoform / epitope: 2 isoforms; check whether the epitope covers both (UniProt).; Regulation: Regulatory changes are unreported (UniProt).. 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.