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ERGIC3 / Endoplasmic reticulum-Golgi intermediate compartment protein 3 · IHC design guide
Design Immunohistochemistry for ERGIC3
ERGIC3 shows broad cytoplasmic tissue staining, with high signal in respiratory epithelium and Purkinje cells (HPA tissue IHC). Use this guide to plan paraffin-section controls and titrate the IHC-validated antibody within 1:100–1:300 (datasheet A09260).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ERGIC3 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 | Observed cytoplasmic (HPA tissue IHC); expected ER–Golgi membranes (UniProt) | |
| Staining pattern | Cytoplasmic in most cells; sparing squamous, stromal and muscle cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A09260) | |
| Positive control | Bronchus+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent across specimens (standard IHC practice). (selected-SKU IHC image A09260) | |
| Caveat | Low staining/RNA concordance; verify tissue signal (HPA tissue IHC) | |
| Regulation | ERGIC2 needed for stable expression (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage across luminal and cytosolic regions is unknown (UniProt) |
Section 1
Recommended ERGIC3 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A09260) with the published ERGIC3 lung cancer IHC protocol (PMC3567939).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A09260) |
| Fixation | Image fixative and duration unreported (datasheet A09260); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A09260); 20 min, 95–100 °C (standard) |
| 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-ERGIC3, 1:100 - 1:300 (datasheet A09260) |
| 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 | ERGIC3-positive staining in respiratory epithelial cells of bronchus (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression in essentially all cells except in squamous epithelial, stroma cells and myocytes. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA retrieval at pH 9.0 (datasheet A09260); the published protocol uses the same retrieval method and pH (PMC3567939).
Section 2
What Is the Expected ERGIC3 Staining Pattern?
ERGIC3 is a membrane protein that cycles between the endoplasmic reticulum and Golgi; its 2 transmembrane segments support an intracellular, cytoplasmic staining pattern in IHC (UniProt Q9Y282 topology and subcellular location). HPA reports widespread cytoplasmic staining, with high signal in selected epithelial, neuronal, trophoblastic and spermatid populations (HPA tissue IHC). Its IHC profile is Approved, but antibody staining has low consistency with RNA expression and awaits external verification (HPA tissue IHC).
| Cytoplasmic staining in bronchial or nasopharyngeal respiratory epithelium, with nearby unstained areas. | This fits a high-signal reference population (HPA: High in respiratory epithelial cells). Assess the intracellular distribution and cell identity together; a brown signal alone does not establish ERGIC3 specificity, especially given the HPA reliability caveat (HPA tissue IHC). |
| Strong, predominantly nuclear or cell-surface staining with little intracellular cytoplasmic signal. | Treat this as a compartment mismatch requiring investigation: ERGIC3 is assigned to ER–Golgi intermediate compartment, cis-Golgi and ER membranes (UniProt Q9Y282 subcellular location). Check the counterstain and whether the apparent signal follows cellular structures before scoring it as positive (general IHC practice). |
| Prominent staining of cardiomyocytes, skeletal myocytes, adipocytes or ovarian stromal cells. | These populations are listed as not detected in the HPA tissue IHC examples (HPA tissue IHC). Reconsider antibody specificity or endogenous detection activity if they dominate the slide; this pattern alone cannot distinguish those causes (general IHC practice). |
| Haze or precipitate across tissue and empty spaces obscures cell boundaries. | A diffuse field-wide signal is difficult to assign to the cytoplasmic pattern reported for ERGIC3 (HPA tissue IHC). Review blocking, washing, chromogen development and detection controls before interpreting cell-specific intensity (general IHC practice). |
| No staining in an adequately preserved bronchial respiratory epithelial control. | HPA reports high staining in this cell population, so absent signal calls for an assay check before interpreting a test section as negative (HPA: High in bronchial respiratory epithelial cells). Confirm that the control contains the intended cells and that primary antibody and detection steps worked (general IHC practice). |
💡Expected ERGIC3 appearanceCall an IHC result positive when identifiable cells show intracellular cytoplasmic staining, potentially concentrated near the ER–Golgi region, with high signal in an HPA high-staining reference population; dominant nuclear, cell-surface or cell-free staining is suspect (UniProt Q9Y282 subcellular location; HPA tissue IHC; general IHC practice).
| Subcellular distribution | ERGIC3 occupies ER–Golgi intermediate compartment, cis-Golgi and ER membranes and cycles between ER and Golgi (UniProt Q9Y282 subcellular location). Interpret a localized intracellular pattern within cytoplasm; the record does not specify one invariant punctate shape for paraffin IHC (UniProt Q9Y282 subcellular location). |
| Reference-cell choice | Respiratory epithelial cells, Purkinje cells, cortical neurons, trophoblasts and elongated or late spermatids have high reported staining; endometrial and appendix glandular cells are medium (HPA tissue IHC). Choose a reference with recognizable cell identity and compare like cells, because intensity differs among listed populations (HPA tissue IHC). |
| Negative and low-staining comparators | HPA lists adipocytes, cardiomyocytes, skeletal and smooth myocytes, ovarian stroma and vaginal squamous epithelium as not detected; some other squamous or glandular populations are low (HPA tissue IHC). These are interpretive comparators, subject to the same HPA reliability caveat (HPA tissue IHC). |
| Antibody and molecular scope | HPA assigns IHC Approved to 2 rabbit polyclonal antibodies, HPA015242 and HPA015968, while reporting low agreement between staining and RNA and pending external verification (HPA antibodies; HPA tissue IHC). UniProt lists 3 isoforms, but the supplied record does not establish which isoforms either antibody detects (UniProt Q9Y282 isoforms). |
| IF/ICC Q&A | Can this IHC pattern serve as an IF/ICC acceptance rule? HPA summarizes ERGIC3 as membrane-associated but supplies no main ICC-IF location, imaged cell lines or ICC validation for these antibodies (HPA subcellular; HPA antibodies). Use the separate IF/ICC guide for that application; do not infer an IF pattern from tissue IHC. |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected positive cells are blank. | The chosen control may lack the intended cell population, or an IHC processing or detection step may have failed (general IHC practice). | Confirm cell identity against an HPA high-staining reference, then check section integrity, retrieval, primary antibody application and detection controls (HPA tissue IHC; general IHC practice). The supplied sources give no ERGIC3-specific retrieval setting. |
| Staining is faint in a low-listed tissue. | The selected population may genuinely have low reported staining, such as esophageal squamous cells or liver cholangiocytes (HPA tissue IHC). | Compare with an HPA high-staining cell population processed in the same IHC run before changing assay conditions (HPA tissue IHC; general IHC practice). Avoid calling weak staining a technical failure from that tissue alone. |
| Muscle, adipose or stromal cells stain strongly. | That conflicts with HPA not-detected examples and may reflect off-target binding or endogenous detection activity (HPA tissue IHC; general IHC practice). | Review a no-primary control and the spatial pattern, and compare another HPA-listed cell population if available (general IHC practice; HPA tissue IHC). Interpret persistent signal cautiously because HPA reports low staining–RNA consistency (HPA tissue IHC). |
| Nuclei dominate the apparent positive signal. | Predominantly nuclear staining does not match the annotated ERGIC3 membrane compartments or HPA cytoplasmic tissue profile (UniProt Q9Y282 subcellular location; HPA tissue IHC). | Check the counterstain and detection controls, then score intracellular cytoplasmic signal separately from nuclear color (general IHC practice). Do not use nuclear intensity alone as evidence of ERGIC3. |
| Diffuse brown background prevents cell-level scoring. | Nonspecific reagent binding, insufficient washing or excessive chromogen development can obscure a cellular IHC pattern (general IHC practice). | Inspect a no-primary control and review blocking, washes and development time under the assay's established IHC conditions (general IHC practice). Resume scoring only when cell boundaries and cytoplasmic signal can be distinguished. |
| An IHC-positive tissue gives an unclear IF/ICC result. | Tissue IHC staining and UniProt localization do not establish antibody performance in IF/ICC; HPA lists no ICC validation or imaged cell lines here (HPA antibodies; HPA subcellular; UniProt Q9Y282 subcellular location). | Evaluate IF/ICC under its own guide and controls. Do not transfer the IHC tissue intensity threshold or infer an IF/ICC protocol from these IHC observations (HPA tissue IHC; HPA subcellular; general IF/ICC practice). |
Experiment evidence
IHC & IF images showing ERGIC3 staining
A09260 · IHCImmunohistochemical analysis of paraffin-embedded human liver cancer. 1, Antibody was diluted at 1:200(4° overnight). 2, Tris-EDTA,pH9.0 was
A09260 · IFImmunofluorescence analysis of HepG2 cells, using ERGI3 Antibody. The picture on the right is blocked with the synthesized peptide.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000125991-ERGIC3/tissue/cerebral+cortex#imid_3921636
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000125991-ERGIC3/tissue/colon#imid_3921578
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000125991-ERGIC3/tissue/liver#imid_3921571
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000125991-ERGIC3/tissue/kidney#imid_3921564
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ERGIC3 IHC & IF
🧪Run bronchus first and score respiratory epithelial cells, which HPA rates High (HPA: bronchus, respiratory epithelial cells High). Use adipose tissue as the negative comparator, scoring adipocytes that HPA rates Not detected; on the bronchus slide, treat adjacent cells without specific staining as internal background comparators without assuming they are ERGIC3-negative (HPA: adipose tissue, adipocytes Not detected; standard IHC control practice).
Positive control tissue: Bronchus (Respiratory epithelial cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for ERGIC3; derive a cell-line control from the positive tissue's cell type (Respiratory epithelial cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and host-matched controls: an isotype control for a monoclonal antibody or nonimmune IgG for a polyclonal antibody; use ERGIC3 knockout material as a biological negative if available (standard IHC control practice). For chromogenic bronchus IHC, quench endogenous peroxidase and check any staining in inflammatory cells against the controls (standard IHC control practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A09260 paraffin-section caption does not state its fixative (selected A09260 tissue-IHC caption). That caption uses Tris-EDTA pH 9.0 retrieval and a 1:200 primary antibody dilution overnight at 4 °C, but it does not establish that retrieval is required under other conditions (selected A09260 tissue-IHC caption). Whether frozen sections or IF/ICC are easier is unreported; for bronchus, assess mucus and inflammatory-cell background during scoring (HPA: bronchus, respiratory epithelial cells High; standard IHC control practice).
HPA tissue IHC evidence for ERGIC3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | Medium | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ERGIC3 IHC Tips
Troubleshoot ERGIC3 staining in paraffin sections by checking retrieval, compartment pattern, controls and cell-specific scoring before interpreting chromogenic signal.
How should I adjust retrieval when ERGIC3 staining is weak?
Start with heat-mediated Tris-EDTA at pH 9.0 for ERGIC3 paraffin sections (datasheet A09260). The selected tissue image used this buffer, a 1:200 primary dilution and incubation at 4°C overnight, so compare retrieval changes while holding those conditions constant (caption A09260). If staining remains weak, compare a shorter heating interval or a citrate buffer at pH 6.0 on serial sections, recording morphology and background alongside signal (standard IHC practice). Include a positive tissue control such as bronchus respiratory epithelium, where ERGIC3 staining is reported as high (HPA: bronchus). Do not interpret stronger diffuse color alone as improved retrieval; ERGIC3 is associated with ER-Golgi membranes (UniProt Q9Y282 subcellular location).
Could fixation explain inconsistent ERGIC3 staining across paraffin blocks?
Target-specific sensitivity of ERGIC3 staining to fixative type or fixation duration is unknown from the supplied evidence; the selected paraffin image does not report a fixative (caption A09260). Record each block’s fixative, time before fixation, fixation duration and processing history before comparing chromogenic intensity (standard IHC practice). For a troubleshooting comparison, stain sections of similar thickness in one run with Tris-EDTA pH 9.0 retrieval and the same 1:200 primary dilution used in the selected image (datasheet A09260; caption A09260). Review morphology and an internal cell population alongside the signal so a processing difference is not mistaken for a biological difference (standard IHC practice).
What staining pattern supports ERGIC3 localisation in tissue?
Expect intracellular cytoplasmic staining compatible with ER-Golgi membranes, including a possible perinuclear concentration, because ERGIC3 cycles between the endoplasmic reticulum and Golgi (UniProt Q9Y282 subcellular location; HPA: tissue profile). Its 2 transmembrane segments place a large lumenal region between residues 47–341, so an isolated nuclear pattern would not fit the annotated topology (UniProt Q9Y282 topology). Compare staining in respiratory epithelial cells, reported high, with nearby stromal cells, which the tissue profile generally excludes (HPA: bronchus; HPA: tissue profile). Check the same cell types in serial sections and inspect whether diffuse chromogen obscures a finer pattern (standard IHC practice). The HPA subcellular summary says “membrane” but supplies no main location or ICC/IF images (HPA: subcellular).
How could isoforms or epitope accessibility affect ERGIC3 IHC?
ERGIC3 has 3 annotated isoforms, but the supplied record does not locate this antibody’s epitope within them (UniProt Q9Y282 isoforms; caption A09260). Request or verify the immunogen coordinates before claiming that a negative section lacks all ERGIC3 isoforms (standard IHC interpretation). The annotated topology puts residues 47–341 in the lumen and the N- and C-termini on the cytoplasmic side; glycosylation sites occur at 241 and 266 (UniProt Q9Y282 topology; UniProt Q9Y282 glycosylation). If staining changes after retrieval, compare matched sections rather than attributing the change to an isoform or glycan without epitope mapping (standard IHC practice). Record the retrieval condition, antibody dilution and cell-specific pattern for each comparison (standard IHC practice).
How can IF help investigate an ambiguous ERGIC3 IHC pattern?
Use IF as a separate check of cellular and compartment localisation; the supplied HPA subcellular entry lists no ICC/IF images (HPA: subcellular). Multiplex ERGIC3 with a marker identifying the expected cell population, such as respiratory epithelial cells in bronchus, and compare signal within those cells with adjacent stroma (HPA: bronchus; HPA: tissue profile). Choose a fluorophore channel after measuring tissue autofluorescence in an unstained section, particularly before judging weak intracellular signal (standard IF practice). Select permeabilisation with the mapped epitope in mind: residues 47–341 are lumenal, whereas residues 1–25 and 363–383 are cytoplasmic (UniProt Q9Y282 topology). Do not transfer the paraffin image’s fixation assumptions to IF, because its fixative is unreported (caption A09260).
How do I reduce diffuse or patchy chromogenic background?
Run a no-primary control and inspect it for endogenous peroxidase activity or nonspecific secondary reagent deposition before changing the ERGIC3 antibody concentration (standard IHC practice). Use a peroxidase block before DAB detection and adjust blocking, washes and chromogen development against that control (standard chromogenic IHC practice). The selected image used primary antibody at 1:200 overnight at 4°C and secondary antibody at 1:200 for 45 minutes at room temperature; those are image conditions, not background-free guarantees (caption A09260). Compare signal in respiratory epithelial cells with nearby stroma, since the HPA tissue profile reports broad cytoplasmic expression but generally excludes stromal cells (HPA: tissue profile; HPA: bronchus).
How should I score heterogeneous ERGIC3 staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring; for example, score bronchus respiratory epithelial cells separately from stroma because their reported ERGIC3 patterns differ (HPA: bronchus; HPA: tissue profile). Record the percentage of positive cells and staining intensity, or calculate an H-score from intensity categories, using identical thresholds across sections (standard IHC quantification practice). Normalise counts to the number of evaluable cells, or report positive-cell density per mm² of viable tissue when cell counting is impractical (standard IHC quantification practice). Exclude folds, section edges and necrotic regions by a prespecified rule, then document retrieval and staining batch for each comparison (standard IHC quantification practice). Report compartment pattern alongside the score, since ERGIC3 is associated with ER-Golgi membranes (UniProt Q9Y282 subcellular location).
When is an ERGIC3-positive IHC result credible?
Treat a cytoplasmic pattern in the expected cells as more credible than isolated nuclear staining: ERGIC3 cycles between ER and Golgi membranes, and HPA describes predominantly cytoplasmic tissue staining (UniProt Q9Y282 subcellular location; HPA: tissue profile). Check a high-staining population such as bronchus respiratory epithelium and a reported undetected population such as cardiomyocytes, while recognising that these references are observational (HPA: bronchus; HPA: heart muscle). Reject edge-only staining, necrotic deposits and signal reproduced by a no-primary control as evidence of specific ERGIC3 localisation (standard IHC interpretation). The HPA tissue assessment is “Approved” but notes low consistency with RNA and pending external verification, so interpret an isolated positive result cautiously (HPA: reliability description).
Boster reagents
Best ERGIC3 / Endoplasmic reticulum-Golgi intermediate compartment protein 3 IHC Antibodies
A09260 has human paraffin-section IHC and HepG2-cell IF images (A09260 IHC/IF captions); human and mouse reactivity is listed (A09260 catalog).
Real IHC data
Anti-ERGI3 ERGIC3 Antibody
Cat # A09260
A09260 will render with an IHC image of paraffin-embedded human liver cancer (A09260 IHC caption). It is listed for IHC, IF and ICC in human and mouse, with an IF image of HepG2 cells (A09260 catalog; A09260 IF caption).
Which to pick: For tissue IHC, choose A09260: its image caption documents paraffin-embedded human liver cancer, Tris-EDTA retrieval at pH 9.0 and primary antibody at 1:200 overnight at 4°C; the fixative is unreported (A09260 IHC caption). For IF/ICC, A09260 is a rabbit polyclonal listed for both applications, with IF shown in HepG2 cells (A09260 catalog; A09260 IF caption). For cross-species work, A09260 lists human and mouse reactivity, although the supplied IHC image shows human material only (A09260 catalog; A09260 IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9Y282 (ERGI3_HUMAN, Endoplasmic reticulum-Golgi intermediate compartment protein 3).
- Human Protein Atlas. ERGIC3 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ERGIC3 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. ERGIC3 antibody validation summary (2 antibodies).
- ERGIC3, which is regulated by miR-203a, is a potential biomarker for non-small cell lung cancer. Cancer science 2015 — PMC4638005.
- Suppression subtractive hybridization identified differentially expressed genes in lung adenocarcinoma: ERGIC3 as a novel lung cancer-related gene. BMC cancer 2013 — PMC3567939.
- PubMed PMID:12747765 — UniProt-cited evidence.
- PubMed PMID:10810093 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ERGIC3 (Endoplasmic reticulum-Golgi intermediate compartment protein 3). Expected localisation Observed cytoplasmic (HPA tissue IHC); expected ER–Golgi membranes (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A09260). Validated IHC antibody: A09260 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Respiratory epithelial cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A09260); 2. Fixation: Image fixative and duration unreported (datasheet A09260); 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: Tris-EDTA pH 9.0 (datasheet A09260); 20 min, 95–100 °C (standard); 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-ERGIC3, 1:100 - 1:300 (datasheet A09260); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent across specimens (standard IHC practice). (selected-SKU IHC image A09260); Caveat: Low staining/RNA concordance; verify tissue signal (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage across luminal and cytosolic regions is unknown (UniProt); Regulation: ERGIC2 needed for stable expression (UniProt). Evidence: UniProt Q9Y282, HPA tissue IHC · Approved, HPA ICC-IF · Membrane, Datasheet A09260, PMC3567939. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ERGIC3/immunohistochemistry-ihc-guide.