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ERGIC1 / Endoplasmic reticulum-Golgi intermediate compartment protein 1 · IHC design guide
Design Immunohistochemistry for ERGIC1
Plan ERGIC1 chromogenic IHC around the cytoplasmic tissue pattern, with strong staining reported in adrenal gland glandular cells and cerebellar Purkinje cells (HPA tissue IHC). Use the catalog antibody at 2–5 μg/ml (datasheet A11007-1) and score cytoplasmic staining by cell type (standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ERGIC1 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 staining (HPA tissue IHC); ER–Golgi membranes expected (UniProt) | |
| Staining pattern | General cytoplasmic staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A11007-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Squamous epithelial cells may stain weakly (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope differences unspecified (UniProt) |
Section 1
Recommended ERGIC1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A11007-1) is accompanied by one published ERGIC1 paraffin-section protocol (PMC5597503 methods).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A11007-1) |
| Fixation | Image fixative and duration unreported (datasheet A11007-1); 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 A11007-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A11007-1) |
| Primary antibody | Rabbit anti-ERGIC1, 2-5 μg/ml (datasheet A11007-1) |
| Primary incubation | Overnight at 4 °C (datasheet A11007-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A11007-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ERGIC1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet A11007-1); the published method does not specify retrieval conditions (PMC5597503 methods).
Section 2
What Is the Expected ERGIC1 Staining Pattern?
ERGIC1 is a two-pass membrane protein that cycles between the endoplasmic reticulum and Golgi (UniProt Q969X5 topology and subcellular location). In paraffin-section IHC, expect broadly cytoplasmic staining rather than a tissue-restricted pattern (HPA: general cytoplasmic expression; low tissue specificity). HPA reports high staining in selected glandular, endocrine, hematopoietic, neuronal and endothelial cells, with an Enhanced tissue-IHC reliability rating (HPA: tissue IHC).
| Cytoplasmic staining in adrenal glandular cells, appendix endocrine cells or cerebellar Purkinje cells. | This fits reported high-staining cell populations (HPA: High in each listed population). Score the relevant cells and cytoplasmic compartment; the tissue name alone is insufficient to judge a slide (HPA: cell-specific tissue IHC). |
| Staining is exclusively nuclear, with little or no cytoplasmic signal, in paraffin-section IHC. | This does not match the reported general cytoplasmic tissue-IHC profile (HPA: tissue IHC). Review morphology, controls and detection background before calling it ERGIC1; nucleoplasmic staining is separately reported for ICC-IF (HPA: subcellular ICC-IF). |
| Strong staining appears mainly in oral or vaginal squamous cells while expected high-staining cells are weak. | Those squamous populations are reported as Low, while selected other populations are High (HPA: tissue IHC). Investigate antibody cross-reactivity or endogenous detection activity using appropriate controls; Low does not mean absent. |
| Brown signal spreads across stroma, lumina or the whole section without clear cellular boundaries. | This is difficult to assign to the reported cytoplasmic cell pattern (HPA: tissue IHC). Diffuse deposit can arise from background in chromogenic IHC (standard IHC practice); compare a negative detection control before scoring cells. |
| No convincing signal appears in a section containing an identified HPA high-staining cell population. | First confirm that the expected cells are present and assess the control slide (HPA: High in the specified cell population). A blank result can reflect assay performance (standard IHC practice); HPA levels do not establish fixation sensitivity. |
💡Expected ERGIC1 appearanceCall a positive result when identifiable HPA high-staining cells show clear cytoplasmic chromogen (HPA: tissue IHC); an isolated nuclear-only pattern or indiscriminate brown background is insufficient for that IHC call (HPA: general cytoplasmic expression; standard IHC practice).
| Cell population and scoring | HPA lists High staining in specific cells, including bone-marrow hematopoietic and colon endothelial cells, but Low staining in soft-tissue fibroblasts (HPA: tissue IHC). Compare like cell populations when judging intensity. |
| Membrane topology and epitope access | ERGIC1 has cytoplasmic residues 1–26 and 276–290 and a lumenal region at 48–254 (UniProt Q969X5 topology). Epitope position could affect access after IHC processing; no antibody epitope or ERGIC1-specific retrieval response is supplied. |
| Isoforms and glycosylation | Three isoforms and one glycosylation site at residue 74 are annotated (UniProt Q969X5). These facts alone cannot predict staining differences or establish that a given antibody detects every isoform; its epitope is unspecified. |
| Antibody evidence | Two rabbit polyclonal antibodies, HPA018666 and HPA018900, have Enhanced IHC validation (HPA: antibody validation). HPA describes medium agreement with RNA and similar staining by paired antibodies (HPA: tissue-IHC reliability); neither makes every stained cell definitive. |
| IF/ICC Q&A: Is nuclear staining expected? | HPA reports supported nucleoplasmic and enhanced vesicular localization in ICC-IF (HPA: subcellular ICC-IF). That observation belongs to cultured-cell IF; interpret paraffin-section IHC against the separate general cytoplasmic tissue profile (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| High-staining reference cells are blank. | The expected cell population may be absent from the section (HPA: cell-specific High staining), or the IHC run may have failed (standard IHC practice). | Identify the reference cells on the counterstain; check a positive control, antibody incubation and detection steps (standard IHC practice). |
| Signal is weak but cellular. | A Low-staining population can be genuinely faint (HPA: oral and vaginal squamous cells; soft-tissue fibroblasts). | Compare the same cell type across sections and include a reported High population as a reference (HPA: tissue IHC); avoid scoring from tissue identity alone. |
| Nuclei dominate the IHC stain. | Nuclear-only IHC conflicts with the general cytoplasmic tissue profile (HPA: tissue IHC), although ICC-IF reports nucleoplasm (HPA: subcellular ICC-IF). | Check controls and cellular boundaries, then repeat interpretation against the tissue-IHC profile; do not transfer the ICC-IF localization call directly to IHC. |
| Brown deposit covers many structures. | Nonspecific background or endogenous detection activity can obscure a cellular chromogenic signal (standard IHC practice). | Review a negative detection control, blocking and wash steps; score only interpretable cytoplasmic staining in identifiable cells (standard IHC practice; HPA: tissue IHC). |
| Unexpected cell types stain more strongly than expected reference cells. | The contrast may reflect background or cross-reactivity; HPA reports cell-specific High and Low levels, with no negative populations listed (HPA: tissue IHC). | Compare matched cell types and controls before assigning specificity. Treat a Low annotation as relative staining, not proof that every positive cell is false (HPA: tissue IHC). |
| Results differ between antibodies or runs. | The supplied record gives Enhanced validation for two antibodies but no epitope mapping or ERGIC1-specific processing sensitivity (HPA: antibody validation; UniProt Q969X5 topology). | Compare staining in the same reported High cell population with matched controls and documented IHC conditions (HPA: tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing ERGIC1 staining
A11007-1 · IHCIHC analysis of ERGIC1 using anti-ERGIC1 antibody (A11007-1). ERGIC1 was detected in a paraffin-embedded section of human liver cancer tissu
A11007-1 · IHCIHC analysis of ERGIC1 using anti-ERGIC1 antibody (A11007-1). ERGIC1 was detected in a paraffin-embedded section of human liver tissue. Heat
A11007-1 · IFIF analysis of ERGIC1 using anti-ERGIC1 antibody (A11007-1). ERGIC1 was detected in an immunocytochemical section of MCF-7 cells. Enzyme ant
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000113719-ERGIC1/tissue/cerebral+cortex#imid_5156420
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000113719-ERGIC1/tissue/colon#imid_5156346
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000113719-ERGIC1/tissue/liver#imid_5156339
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000113719-ERGIC1/tissue/kidney#imid_5156340
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018666; green = ERGIC1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018666; green = ERGIC1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ERGIC1 IHC & IF
🧪Run adrenal gland first and look for staining in glandular cells (HPA: High in adrenal gland glandular cells). HPA detects ERGIC1 in all 45 scored tissues, so use no-primary and isotype controls in place of a negative tissue; cells without specific DAB signal should show counterstain alone, but no named cell population on that slide is established as ERGIC1-negative (HPA: no negative tissue rows).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: ERGIC1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ERGIC1 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (supported), Vesicles (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide, a control immunoglobulin matched to the primary antibody’s host species and class, and, if available, ERGIC1 knockout material or a validated peptide-block control (selected-SKU caption: rabbit primary and anti-rabbit secondary). On adrenal sections, quench endogenous peroxidase and check blood-rich areas for nonspecific DAB signal (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU tissue-IHC caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 is documented for a paraffin section of liver cancer, but its necessity for adrenal gland has not been established (selected-SKU tissue-IHC caption: EDTA pH 8.0). HPA provides ICC-IF images, but the supplied evidence does not establish that IF or frozen sections are easier than paraffin IHC; assess any blood-associated DAB signal against the peroxidase control (HPA: ICC-IF images in A-431, U-251MG and U2OS; standard chromogenic IHC practice).
HPA tissue IHC evidence for ERGIC1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Paired antibodies with high similarity support the protein profile.). 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 | Endocrine cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: ERGIC1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced ERGIC1 IHC Tips
Troubleshoot ERGIC1 staining in paraffin sections by checking retrieval, cellular distribution, controls, and cell-type-specific scoring.
Where should I start if ERGIC1 staining is weak in paraffin sections?
Begin with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A11007-1). The selected tissue-IHC image used this retrieval before incubation with 2 μg/ml antibody overnight at 4°C (caption A11007-1). If staining remains weak, check that heating, cooling, and reagent coverage were consistent across sections before changing the retrieval conditions (standard IHC practice). Compare any retrieval adjustment on adjacent sections with the same detection and exposure conditions, because stronger DAB deposition alone does not establish greater specificity (standard IHC practice). Evaluate signal in the expected cytoplasmic distribution and in a documented positive cell population (HPA tissue IHC: general cytoplasmic expression; High in adrenal gland glandular cells).
Can fixation explain variable ERGIC1 staining between tissue blocks?
Target-specific fixation sensitivity is unknown: the selected ERGIC1 caption identifies a paraffin-embedded section but does not state its fixative (caption A11007-1). Record each block's fixative and processing history, then compare sections with similar handling before attributing a staining difference to ERGIC1 biology (standard IHC practice). Fixation can alter epitope accessibility in IHC generally, so assess retrieval and staining controls alongside the specimen when processing histories differ (standard IHC practice). Keep EDTA retrieval at pH 8.0 as the documented starting condition for this antibody (datasheet A11007-1). Neither the HPA tissue pattern nor ERGIC1's membrane topology establishes a fixation-specific effect for this target (HPA tissue IHC; UniProt Q969X5 topology).
What cellular pattern should I expect in ERGIC1 chromogenic IHC?
Start by assessing cytoplasmic staining, the general pattern reported in tissue IHC (HPA tissue IHC: general cytoplasmic expression). ERGIC1 is annotated on endoplasmic reticulum, ER–Golgi intermediate compartment, and Golgi membranes and cycles between the endoplasmic reticulum and Golgi (UniProt Q969X5 subcellular). At light-microscope resolution, judge whether intracellular DAB signal is plausible in the relevant cell population rather than demanding visibly resolved individual membranes (standard IHC interpretation; UniProt Q969X5 subcellular). HPA cell imaging also reports nucleoplasm and vesicles, so a nuclear component warrants comparison with independent controls instead of automatic rejection (HPA subcellular: nucleoplasm supported, vesicles enhanced). Compare similarly processed sections and document cytoplasmic and nuclear signal separately (standard IHC practice).
Could ERGIC1 isoforms or epitope position change my IHC result?
ERGIC1 has 3 annotated isoforms, but the supplied antibody caption does not identify its epitope or establish isoform coverage (UniProt Q969X5 isoforms; caption A11007-1). The reference protein has transmembrane segments at residues 27–47 and 255–275, with a lumenal region at 48–254 (UniProt Q969X5 topology). It also has one annotated glycosylation site at residue 74; these annotations alone cannot predict this antibody's staining response (UniProt Q969X5 glycosylation; standard IHC interpretation). If two samples differ after matched retrieval and detection, check whether independent epitope information or an orthogonal assay supports the difference before calling it isoform-specific (standard IHC practice). Report the observed cellular pattern without assigning an individual isoform (UniProt Q969X5 isoforms; caption A11007-1).
How should I assess ERGIC1 localisation in multiplex IF?
For multiplex IF, pair ERGIC1 with a marker that identifies the cell population being examined, such as Purkinje cells where HPA reports high tissue-IHC staining (HPA tissue IHC: High in cerebellar Purkinje cells; standard IF practice). Choose well-separated fluorophores and assess tissue autofluorescence in each channel before interpreting overlap (standard IF practice). ERGIC1 spans membranes twice, with cytoplasmic ends and a lumenal region, so permeabilisation must permit access to the antibody's epitope side (UniProt Q969X5 topology; standard IF practice). Because the supplied caption does not map that epitope, test accessibility rather than assuming which compartment the antibody detects (caption A11007-1). Compare any vesicular or nuclear signal with the reported cell-imaging locations and channel controls (HPA subcellular: vesicles enhanced, nucleoplasm supported; standard IF practice).
How can I separate ERGIC1 signal from chromogenic background?
First compare staining in the expected cytoplasmic compartment with DAB deposition at tissue edges, damaged areas, and blank regions (HPA tissue IHC: general cytoplasmic expression; standard IHC practice). The selected caption used 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (caption A11007-1). If background is widespread, check blocking, washes, primary-antibody concentration, and chromogen development while keeping adjacent sections comparable (standard IHC practice). Include a no-primary control to reveal detection-related signal, and use an endogenous-peroxidase block when applying HRP/DAB detection (standard IHC practice). The caption documents HRP-linked detection and DAB, whereas peroxidase blocking is a general workflow control (caption A11007-1; standard IHC practice).
What is a defensible way to score ERGIC1 IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic scoring compartment before reading slides, because HPA describes general cytoplasmic staining with differing levels among cell types (HPA tissue IHC). Record the percentage of positive cells and staining intensity, or calculate an H-score using intensity grades 0–3 and the percentage at each grade (standard IHC scoring practice). For spatial analyses, report positive-cell density per mm² of evaluable tissue and exclude non-evaluable regions by a prespecified rule (standard IHC scoring practice). Normalize comparisons to the same eligible cell population or tissue area, rather than whole-slide DAB coverage when cellular composition differs (standard IHC scoring practice). Apply identical retrieval, detection, and scoring thresholds across compared sections (standard IHC practice).
Which findings support true ERGIC1 staining rather than artefact?
A convincing result shows reproducible intracellular staining in the scored cells, consistent with the general cytoplasmic tissue profile and ER–Golgi membrane annotation (HPA tissue IHC; UniProt Q969X5 subcellular). High signal in adrenal gland glandular cells or cerebellar Purkinje cells is supported by HPA, but cell identity and section quality still need confirmation (HPA tissue IHC: High in both populations; standard IHC interpretation). Treat edge staining, necrotic deposits, and signal retained in a no-primary control as possible artefacts (standard IHC practice). Endogenous peroxidase can contribute DAB signal, so inspect the relevant control before interpreting strong chromogen deposits (standard IHC practice). Predominantly nuclear staining requires separate validation: cell imaging reports nucleoplasm, while tissue IHC reports general cytoplasmic expression (HPA subcellular; HPA tissue IHC).
Boster reagents
Best ERGIC1 / Endoplasmic reticulum-Golgi intermediate compartment protein 1 IHC Antibodies
A11007-1 has IHC images from human liver cancer and liver paraffin sections (catalog IHC captions) and an IF image from MCF-7 cells (catalog IF caption).
Real IHC data
Anti-ERGIC1 Antibody ®
Cat # A11007-1
A11007-1 will render with its human liver cancer paraffin-section IHC figure (catalog IHC caption). The same SKU has a human liver paraffin-section IHC image and an MCF-7 cell IF image (catalog IHC and IF captions).
Which to pick: Choose A11007-1 for paraffin-section IHC: its human liver cancer caption documents EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (catalog IHC caption); the fixative is unreported (catalog IHC caption). For IF/ICC, the same SKU has an MCF-7 cell IF image at 5 μg/ml (catalog IF caption). For cross-species work, A11007-1 lists human, mouse and rat reactivity (catalog applications and reactivity), while its supplied IHC images show human tissue (catalog IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q969X5 (ERGI1_HUMAN, Endoplasmic reticulum-Golgi intermediate compartment protein 1).
- Human Protein Atlas. ERGIC1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ERGIC1 subcellular location (ICC-IF): Localized to the nucleoplasm and vesicles..
- Human Protein Atlas. ERGIC1 antibody validation summary (2 antibodies).
- Aberrant DNA-PKcs and ERGIC1 expression may be involved in initiation of gastric cancer. World journal of gastroenterology 2017 — PMC5597503.
- PubMed PMID:10574461 — UniProt-cited evidence.
- PubMed PMID:10931946 — UniProt-cited evidence.
- PubMed PMID:11230166 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ERGIC1 (Endoplasmic reticulum-Golgi intermediate compartment protein 1). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); ER–Golgi membranes expected (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A11007-1). Validated IHC antibody: A11007-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A11007-1); 2. Fixation: Image fixative and duration unreported (datasheet A11007-1); 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 A11007-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A11007-1); 8. Primary antibody: Rabbit anti-ERGIC1, 2-5 μg/ml (datasheet A11007-1); 9. Primary incubation: Overnight at 4 °C (datasheet A11007-1). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Squamous epithelial cells may stain weakly (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope differences unspecified (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q969X5, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm and vesicles., Datasheet A11007-1, PMC5597503. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ERGIC1/immunohistochemistry-ihc-guide.