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COPB2 / Coatomer subunit beta' · IHC design guide
Design Immunohistochemistry for COPB2
Plan chromogenic COPB2 IHC in paraffin sections using the catalog antibody at 1:50–1:200 (datasheet A07493). Colon glandular cells show high staining, while adipocytes have no detected staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
COPB2 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 | General cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in many tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, heat-mediated (datasheet A07493) | |
| Positive control | Bronchus+4 more · see all | |
| Negative control | Adipose tissue+3 more · see all |
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 | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended COPB2 IHC & IF Protocols
The catalog antibody protocol (datasheet: A07493) is accompanied by published COPB2 IHC protocols for gastric cancer, hepatocellular carcinoma, and gastric cancer xenograft sections (PMC12867441; PMC11964868; PMC6411345).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A07493) |
| Fixation | Image fixative and duration unreported (datasheet A07493); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6.0 (datasheet A07493); 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-COPB2, 1:50-1:200 (datasheet A07493) |
| 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 | COPB2-positive staining in respiratory epithelial cells of bronchus (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 heat retrieval (datasheet: A07493); a gastric cancer TMA study also used citrate pH 6.0 (PMC12867441).
Section 2
What Is the Expected COPB2 Staining Pattern?
COPB2 should produce cytoplasmic staining in many cell types, with high staining documented in bronchial respiratory epithelium and colon, duodenal, and pancreatic glandular cells (HPA tissue IHC: general cytoplasmic expression; High in these cells). It is cytosolic and associates with the cytoplasmic face of Golgi and COPI vesicle membranes; it has no transmembrane segment (UniProt P35606 topology). The HPA tissue IHC assessment is Approved, with medium consistency between staining and RNA expression (HPA tissue IHC: reliability).
| Cytoplasmic chromogen in bronchial respiratory epithelial cells or colon glandular cells. | This matches the reported high staining in those cells (HPA tissue IHC: High). Judge the signal within the identified cell population: COPB2 is associated with cytosol, Golgi membranes, and COPI vesicles (UniProt P35606 subcellular location). A visibly concentrated Golgi region can be compatible, but tissue IHC does not establish a required Golgi pattern (HPA tissue IHC: general cytoplasmic expression). |
| Signal appears confined to nuclei or cell surfaces, without convincing cytoplasmic staining. | Treat that distribution as discordant with the reported general cytoplasmic tissue pattern (HPA tissue IHC: profile) and cytosolic or cytoplasmic membrane association (UniProt P35606 subcellular location; topology). Review morphology and control staining before assigning it to COPB2; localization alone cannot identify the source of an unexpected signal. |
| Strong staining in adipocytes or ovarian stromal cells while expected epithelial cells remain weak. | These cell populations are reported as not detected, whereas several epithelial and glandular populations are High (HPA tissue IHC: adipose tissue, ovary, bronchus, colon). Check cell identification and detection controls; cross-reactivity or endogenous detection activity are possible explanations under standard IHC practice. An unexpected signal does not by itself prove either cause. |
| Haze or chromogen covers stroma and empty spaces, obscuring cell boundaries. | A field-wide deposit is difficult to reconcile with a cell-resolved cytoplasmic pattern (HPA tissue IHC: general cytoplasmic expression). Under standard IHC practice, assess blocking, antibody concentration, washing, and chromogen development with an appropriate negative detection control. Do not score obscured cells as positive. |
| No detectable signal in bronchial respiratory epithelium or pancreatic exocrine glandular cells. | Both are reported High, so a blank result warrants a technical check (HPA tissue IHC: bronchus, pancreas). Confirm that the selected antibody is validated for IHC and that the detection controls worked (HPA antibodies: HPA036867 IHC Approved). The HPA reliability rating is Approved with medium RNA–staining consistency, so a single negative section cannot establish true absence (HPA tissue IHC: reliability). |
💡Expected COPB2 appearanceCall a result positive when identifiable respiratory epithelial or glandular cells show convincing cytoplasmic chromogen, potentially with regional concentration, at an intensity consistent with HPA High examples; isolated nuclear, surface-only, or field-wide staining is discordant (HPA tissue IHC: profile and High cells; UniProt P35606 subcellular location).
| Compartment assignment | COPB2 is cytosolic or bound on the cytoplasmic side of Golgi and COPI vesicle membranes, with no transmembrane segment (UniProt P35606 topology; subcellular location). Interpret a cytoplasmic signal in that context. The supplied evidence does not define an antibody epitope, so it cannot support a claim that a particular retrieval condition exposes it. |
| Choice of reference tissue and cell population | Bronchial respiratory epithelial, colon and duodenal glandular, and pancreatic exocrine glandular cells are High; adipocytes, ovarian stromal cells, smooth muscle cells, and splenic red-pulp cells are Not detected (HPA tissue IHC). Compare the same cell populations when interpreting a section; a tissue name alone can conceal different cell-level results. |
| Evidence strength for tissue IHC | The tissue profile is Approved but has medium consistency with RNA expression data, and RNA specificity is low across tissues (HPA tissue IHC: reliability; RNA specificity). Use the reported cell-level observations as comparators, while allowing for uncertainty in an individual specimen. This evidence supplies no target-specific fixation or antigen-retrieval effect. |
| Assay-specific antibody validation | HPA036867 is listed as IHC Approved, while HPA058180 is listed as ICC Approved without an IHC status in the supplied record (HPA antibodies). An ICC result does not establish IHC performance. The supplied antibody record gives no dilution, retrieval setting, or epitope for this section. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known High epithelial cells are blank. | The result conflicts with the HPA High observations; the supplied data do not identify a COPB2-specific technical cause (HPA tissue IHC: bronchus, colon, pancreas). | Check the IHC antibody's validation status, section integrity, and positive detection control; then review the established staining workflow (HPA antibodies: HPA036867 IHC Approved; standard IHC practice). |
| Nearly every cell and tissue space stains. | Diffuse deposit prevents assessment of the reported cell-resolved cytoplasmic pattern (HPA tissue IHC: general cytoplasmic expression). Background from detection or chromogen handling is possible under standard IHC practice. | Run a negative detection control and review blocking, washes, antibody concentration, and development time (standard IHC practice). Re-score only cells with interpretable morphology. |
| Staining is mainly nuclear or at the cell surface. | That distribution disagrees with COPB2's cytosolic and cytoplasmic-face membrane locations (UniProt P35606 subcellular location; topology). Its origin cannot be determined from appearance alone. | Check cell boundaries and counterstain, compare a documented High tissue, and inspect negative controls before assigning specificity (HPA tissue IHC: High examples; standard IHC practice). |
| Adipocytes, smooth muscle, or ovarian stromal cells appear strongly positive. | HPA reports these cell populations as Not detected (HPA tissue IHC: adipose tissue, smooth muscle, ovary). Misidentified cells, cross-reactivity, or endogenous detection activity are possible under standard IHC practice. | Verify cell identity and compare negative detection controls with nearby expected positive cells; report discordant staining without calling its cause proven (standard IHC practice; HPA tissue IHC: High epithelial and glandular cells). |
| The same tissue contains both strong and blank areas. | HPA results are cell-specific, and its overall tissue IHC assessment has medium RNA–staining consistency (HPA tissue IHC: cell levels; reliability). Tissue-wide scoring can therefore obscure which cells carry the signal. | Score identifiable cell populations separately, using morphology and a counterstain; compare each population with its HPA entry (standard IHC practice; HPA tissue IHC: cell-level observations). |
| Why does ICC-IF show an ER-associated signal alongside Golgi staining? | HPA ICC-IF lists the ER as the approved main location and Golgi as an approved additional location; UniProt describes cytosol and the cytoplasmic side of Golgi and COPI vesicles (HPA subcellular ICC-IF; UniProt P35606 subcellular location). | Interpret the ICC-IF result within its own assay and antibody validation: HPA058180 is ICC Approved, while HPA036867 is IHC Approved (HPA antibodies). Use the tissue IHC profile for chromogenic-section expectations (HPA tissue IHC: general cytoplasmic expression). |
Experiment evidence
IHC & IF images showing COPB2 staining
A07493 · IHCImmunohistochemistry of paraffin-embedded human breast cancer using COPB2 Rabbit pAb at dilution of 1:50 .Perform high pressure antigen retr
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000184432-COPB2/tissue/cerebral+cortex#imid_8956298
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000184432-COPB2/tissue/colon#imid_8956216
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000184432-COPB2/tissue/liver#imid_8956225
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000184432-COPB2/tissue/kidney#imid_8956218
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA058180; green = COPB2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA058180; green = COPB2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for COPB2 IHC & IF
🧪Start with bronchus: respiratory epithelial cells should stain (HPA: High in bronchus respiratory epithelial cells). Use adipose tissue as the negative comparator (HPA: adipocytes Not detected); on the bronchus slide, assess any unstained neighboring cells for background-only signal without assuming they are COPB2-negative.
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 ICC-IF images show COPB2 in A-431, HeLa, U2OS, with annotated localisation: Endoplasmic reticulum (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) section, a species-matched nonimmune rabbit IgG isotype control for the catalog rabbit polyclonal antibody (caption: Rabbit pAb), and COPB2-knockout material processed alongside the test section if available (standard IHC practice). Quench endogenous peroxidase and check mucus-associated background in bronchus before interpreting chromogenic staining (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected A07493 paraffin-section caption does not state the fixative (caption: fixative not stated). The caption uses high-pressure retrieval in 10 mM citrate, pH 6.0, at 1:50 in breast cancer tissue, which provides a starting condition but does not establish that retrieval is required for bronchus (caption: A07493 tissue IHC). IF/ICC localization can be assessed in the HPA-imaged A-431, HeLa, or U2OS cells (HPA: ICC-IF image cell lines); the supplied evidence does not establish that IF or frozen sections are easier than paraffin IHC, and bronchial mucus should be checked for staining artefact (standard IHC practice).
HPA tissue IHC evidence for COPB2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced COPB2 IHC Tips
Use compartment, cell type, and matched controls to evaluate COPB2 staining in paraffin sections; treat fixation effects and epitope specificity as unresolved.
What retrieval should I use when COPB2 staining is weak in paraffin sections?
Start with heat-mediated retrieval in 10 mM citrate buffer, pH 6.0 (datasheet A07493). The selected paraffin-section image used high-pressure retrieval with that buffer and a 1:50 antibody dilution (A07493 caption). If staining is weak, compare longer and shorter heating within the citrate condition on adjacent sections, while keeping detection settings and section thickness constant (standard IHC practice). Record tissue integrity alongside signal, since excessive heating can damage morphology and make cytoplasmic staining harder to score (standard IHC practice). Include a no-primary control to distinguish weak COPB2 signal from chromogen background (standard IHC practice).
How should I investigate possible fixation effects on COPB2 IHC?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (A07493 caption). Do not assign a formalin exposure or attribute weak staining to COPB2-specific crosslinking from that image (A07493 caption). Compare sections with documented fixation histories using the same 10 mM citrate, pH 6.0 retrieval and detection run (datasheet A07493; standard IHC practice). Inspect morphology and staining in matched cell populations before changing retrieval or antibody concentration (standard IHC practice). Keep fixation time, processing, section thickness, and run conditions in the record so any observed difference can be assessed without assuming a target-specific effect (standard IHC practice).
Where should credible COPB2 chromogenic staining appear?
Expect predominantly cytoplasmic staining, with possible Golgi-associated concentration, because COPB2 is cytosolic and associates with the cytoplasmic face of Golgi and COPI-coated vesicle membranes (UniProt P35606 localisation). HPA tissue IHC describes general cytoplasmic expression, while its cellular imaging reports endoplasmic reticulum and Golgi localisation (HPA tissue IHC; HPA subcellular). Chromogenic sections may not resolve those organelles, so judge the overall cytoplasmic pattern rather than requiring discrete puncta (standard IHC practice). COPB2 has no annotated transmembrane segment, making an isolated cell-surface rim difficult to reconcile with its recorded topology (UniProt P35606 topology). Compare the pattern with matched control sections before calling an unusual compartment positive (standard IHC practice).
Can this antibody distinguish COPB2 isoforms or modified epitopes in tissue?
COPB2 has 2 listed isoforms, but the supplied product caption gives no antibody epitope or isoform coverage (UniProt P35606 isoforms; A07493 caption). Its recorded modifications include acetyllysine 627, phosphoserine 859, and phosphothreonine 861; their relationship to this antibody's binding is unreported (UniProt P35606 modified residues; A07493 caption). Treat chromogenic staining as COPB2-associated rather than as evidence for a particular isoform or modification (standard IHC interpretation). If those distinctions matter, obtain epitope mapping and use independently validated modification or isoform-specific controls before assigning them to stained cells (standard IHC practice). Keep retrieval and detection matched across comparisons so technical variation does not masquerade as epitope selectivity (standard IHC practice).
How could IF help assess an ambiguous COPB2 IHC pattern?
Use IF as a complementary localisation check for an ambiguous chromogenic pattern, with a marker identifying the expected cell population in a separate channel (standard IF practice). For an epithelial IHC region, compare COPB2 with that region's cell marker; HPA reports high staining in bronchial respiratory epithelial and colon glandular cells (HPA tissue IHC). Choose fluorophores after checking tissue autofluorescence and single-channel controls, especially where broad background could obscure a perinuclear signal (standard IF practice). Permeabilise sufficiently for an antibody to access cytosolic COPB2 on the cytoplasmic side of Golgi-associated membranes (UniProt P35606 topology; standard IF practice). Assess IF and IHC separately because the A07493 caption documents paraffin-section IHC, not an IF protocol (A07493 caption).
How can I separate COPB2 staining from chromogenic background?
Run a no-primary section through the same secondary reagent and chromogen steps to reveal detection-system background (standard IHC practice). For peroxidase-based detection, include a peroxidase block and inspect the control before interpreting brown cytoplasmic deposits as COPB2 (standard IHC practice). Titrate around the caption's 1:50 dilution while keeping citrate retrieval at pH 6.0 and development conditions matched (A07493 caption; standard IHC practice). HPA reports high staining in colon glandular cells and no detected staining in adipocytes, which can inform cell-level comparison but do not replace controls on the tested sections (HPA tissue IHC; standard IHC practice). Flag pigment, folds, and tissue edges separately during review (standard IHC practice).
What scoring method makes COPB2 IHC comparable across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population before analysis, then report the percentage of positive cells and an intensity-based H-score for cytoplasmic staining (standard IHC practice; HPA tissue IHC profile). If spatial distribution matters, report positive-cell density per mm² of viable tissue alongside the cell-based score (standard IHC practice). Normalise counts to the same annotated viable area and compare like cell populations, rather than whole sections with different cellular composition (standard IHC practice). Set thresholds from matched control staining and hold retrieval, antibody dilution, chromogen development, and imaging settings constant (standard IHC practice). Report the 1:50 dilution separately when using the pictured A07493 condition (A07493 caption).
When should an apparent COPB2-positive region be treated as artefact?
A cytoplasmic pattern in the relevant cells is more plausible than isolated nuclear or cell-surface staining, given COPB2's cytosolic and Golgi-associated localisation (UniProt P35606 localisation and topology; HPA tissue IHC). Compare cell identity with HPA's high staining in bronchial respiratory epithelial cells and colon glandular cells, while recognising its reported medium consistency with RNA data (HPA tissue IHC). Treat staining confined to tissue edges, folds, or necrotic regions as suspect until it persists in intact interior tissue (standard IHC practice). Check no-primary and peroxidase-block controls for endogenous enzyme or detection background before assigning DAB deposits to COPB2 (standard IHC practice). Confirm unexpected patterns on independently processed sections (standard IHC practice).
Boster reagents
Best COPB2 / Coatomer subunit beta' IHC Antibodies
A07493 has real COPB2 IHC data from paraffin-embedded human breast cancer (IHC image caption). Its catalog lists IF/ICC and human, mouse, and rat reactivity (catalog).
Real IHC data
Anti-Coatomer subunit beta' COPB2 Antibody
Cat # A07493
A07493 will render with an IHC image of paraffin-embedded human breast cancer stained at 1:50 after citrate retrieval (IHC image caption). IF/ICC and human, mouse, and rat reactivity are catalog-listed; no IF image is supplied (catalog payload).
Which to pick: For tissue IHC, choose A07493 for paraffin sections: its image shows human breast cancer after high-pressure retrieval in 10 mM citrate buffer at pH 6.0; the fixative is unreported (IHC image caption). For IF/ICC, A07493 is listed at 1:50–1:100, but has no supplied IF image (catalog payload). A07493 is a rabbit polyclonal listed as reactive with human, mouse, and rat; the supplied IHC image demonstrates staining only in human breast cancer (catalog; IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P35606 (COPB2_HUMAN, Coatomer subunit beta').
- Human Protein Atlas. COPB2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. COPB2 subcellular location (ICC-IF): Mainly localized to the endoplasmic reticulum. In addition localized to the Golgi apparatus..
- Human Protein Atlas. COPB2 antibody validation summary (2 antibodies).
- COPB2 drives gastric cancer progression via PI3K/AKT/NF-κB signaling: a multi-omics and functional study. Cell adhesion & migration 2026 — PMC12867441.
- COPB2 gene silencing inhibits colorectal cancer cell proliferation and induces apoptosis via the JNK/c-Jun signaling pathway. PloS one 2020 — PMC7676692.
- COPB2 promotes hepatocellular carcinoma progression through regulation of YAP1 nuclear translocation. Oncology research 2025 — PMC11964868.
- Silencing of COPB2 inhibits the proliferation of gastric cancer cells and induces apoptosis via suppression of the RTK signaling pathway. International journal of oncology 2019 — PMC6411345.
- PubMed PMID:8335000 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16641997 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for COPB2 (Coatomer subunit beta'). Expected localisation General cytoplasmic staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, heat-mediated (datasheet A07493). Validated IHC antibody: A07493 at 1:50-1:200 (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 breast cancer tissue; fixative not specified (datasheet A07493); 2. Fixation: Image fixative and duration unreported (datasheet A07493); 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: Citrate pH 6.0 (datasheet A07493); 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-COPB2, 1:50-1:200 (datasheet A07493); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). Evidence: UniProt P35606, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the endoplasmic reticulum. In addition localized…, Datasheet A07493, PMC12867441, PMC11964868. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/COPB2/immunohistochemistry-ihc-guide.