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COPB2 / Coatomer subunit beta' · IHC design guide
Design Immunohistochemistry for COPB2
Plan chromogenic COPB2 IHC in paraffin sections with the catalog antibody at 1:50–1:200 (datasheet). Assess cytoplasmic staining against high-staining glandular cells and undetected adipocytes (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 | Cytoplasmic staining in tissue (HPA tissue IHC); Golgi and vesicle association (UniProt) | |
| Staining pattern | General cytoplasmic staining; glandular cells can stain strongly (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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07493) | |
| Caveat | Antibody staining has medium consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Cell-level intensity varies (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; epitope coverage needs validation (UniProt) |
Section 1
Recommended COPB2 IHC & IF Protocols
The catalog antibody protocol uses citrate pH 6.0 retrieval (datasheet A07493). The published COPB2 IHC protocols below cover gastric cancer, hepatocellular carcinoma, and 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 for the catalog antibody (datasheet A07493); the hepatocellular carcinoma study instead reports alkaline EDTA retrieval (PMC11964868).
Section 2
What Is the Expected COPB2 Staining Pattern?
COPB2 should show mainly cytoplasmic staining in paraffin sections, with Golgi associated signal consistent with a cytosolic coatomer on the cytoplasmic face of Golgi and COPI vesicle membranes (UniProt P35606 topology and subcellular location). Glandular and respiratory epithelial cells provide strong examples (HPA: High in listed cells). HPA describes general cytoplasmic expression and rates its tissue IHC profile Approved, with medium consistency against RNA data (HPA: tissue IHC).
| Strong cytoplasmic staining in colon or duodenal glandular cells, or bronchial respiratory epithelial cells (HPA: High in these cells). | This matches the reported cell types and broad cytoplasmic pattern (HPA: tissue IHC). A Golgi associated component is plausible because coatomer associates with Golgi membranes and COPI vesicles (UniProt P35606 subcellular location). Judge the signal within the appropriate cells, alongside the section's controls (general IHC practice). |
| Predominantly nuclear staining, with little cytoplasmic signal (HPA: general cytoplasmic expression; UniProt P35606 subcellular location). | This is a compartment mismatch: neither source places COPB2 mainly in nuclei (HPA: tissue IHC; UniProt P35606 subcellular location). Treat a nuclear dominant result as suspect and check whether it persists in a no primary control before interpreting it as COPB2 (general IHC practice). |
| Strong staining in adipocytes, smooth muscle cells, ovarian stroma, or splenic red pulp cells (HPA: Not detected in these cells). | That conflicts with the listed HPA tissue observations and warrants a specificity check (HPA: tissue IHC). Cross reactivity or endogenous detection activity are possible explanations, not diagnoses from appearance alone; compare cell type matched controls and the no primary control (general IHC practice). |
| Brown precipitate covers tissue broadly, including areas without a convincing cell specific cytoplasmic pattern (HPA: general cytoplasmic expression). | Diffuse deposition obscures the reported cellular pattern (HPA: tissue IHC). Background can arise from detection reagents, inadequate blocking, or concentrated antibody (general IHC practice). Inspect the no primary control and adjust detection or blocking before scoring cells (general IHC practice). |
| No stain in colon glandular cells or bronchial respiratory epithelial cells (HPA: High in these cells). | This fails an expected positive comparison but does not by itself establish absence of COPB2 (HPA: tissue IHC; general IHC practice). Check section quality, retrieval and staining controls, antibody dilution, and detection performance (general IHC practice). No COPB2 specific retrieval or fixation sensitivity is supplied. |
💡Expected COPB2 appearanceCall a positive result when the appropriate glandular or respiratory epithelial cells show strong, mainly cytoplasmic staining, potentially enriched around the Golgi (HPA: High in listed cells and general cytoplasmic expression; UniProt P35606 subcellular location); predominantly nuclear or indiscriminate stromal staining is suspect (HPA: tissue IHC; general IHC practice).
| Which cell types provide contrast? | Colon and duodenal glandular cells and bronchial respiratory epithelial cells are reported High; adipocytes and smooth muscle cells are Not detected (HPA: tissue IHC). Compare the named cell types rather than treating every cell in a tissue as equivalent (general IHC practice). |
| How much weight should the tissue pattern carry? | The tissue IHC profile is Approved, with medium consistency between antibody staining and RNA expression data (HPA: reliability). Use its High and Not detected calls as reference observations, while checking the actual cell type and local controls (HPA: tissue IHC; general IHC practice). |
| What compartment should guide scoring? | HPA reports general cytoplasmic staining; UniProt places coatomer in cytosol and on the cytoplasmic side of Golgi and COPI vesicle membranes, without a transmembrane segment (HPA: tissue IHC; UniProt P35606 topology and subcellular location). |
| Do isoforms or modifications change the prediction? | UniProt lists two isoforms and annotated acetylation and phosphorylation sites (UniProt P35606 isoforms and modified residues). The payload gives no antibody epitope or isoform specific IHC pattern, so these annotations do not justify a different slide prediction. |
| What does IF/ICC add? | HPA ICC-IF reports mainly endoplasmic reticulum and additional Golgi localization, using cell images from A-431, HeLa, and U2OS (HPA: subcellular). Treat this as a separate imaging observation; it does not replace the paraffin tissue IHC pattern (HPA: tissue IHC and subcellular). |
| Can antibody validation be transferred between assays? | HPA036867 is Approved for IHC, while HPA058180 is Approved for ICC; the supplied statuses do not validate either antibody for the other's assay (HPA: antibodies). Interpret the paraffin result using IHC controls (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected High cells are blank while the section counterstains normally (HPA: High in listed glandular and respiratory epithelial cells). | Primary antibody, retrieval, or detection may have failed; a counterstain alone does not verify the immunostain (general IHC practice). No COPB2 specific retrieval condition is supplied. | Check a known positive section in the same run, reagent order, antibody dilution, retrieval conditions, and detection controls (HPA: High reference cells; general IHC practice). |
| Strong nuclear signal dominates the slide (HPA: general cytoplasmic expression). | The location conflicts with HPA tissue IHC and UniProt cytosol/Golgi placement; nonspecific binding or detection artefact is possible (HPA: tissue IHC; UniProt P35606 subcellular location; general IHC practice). | Compare the no primary control, check detection background, and score only a reproducible cytoplasmic pattern in the expected cells (HPA: tissue IHC; general IHC practice). |
| Brown signal appears in adipocytes or smooth muscle cells (HPA: Not detected in these cells). | The cell type contrast disagrees with HPA; cross reactivity or endogenous enzyme activity could contribute (HPA: tissue IHC; general IHC practice). | Identify the stained cells, inspect the no primary control, and address endogenous detection activity according to the detection chemistry (general IHC practice). |
| Diffuse haze makes cytoplasmic staining hard to resolve (HPA: general cytoplasmic expression). | High background may reflect antibody concentration, blocking, or detection conditions (general IHC practice). The haze alone cannot identify which component is responsible. | Compare control and stained sections, then adjust antibody dilution, blocking, or washing one variable at a time (general IHC practice). |
| Perinuclear or Golgi associated stain is present, but a broad cytoplasmic component is weak (HPA: general cytoplasmic expression; UniProt P35606 subcellular location). | Golgi association is biologically plausible, yet intensity and distribution depend on the observed preparation; the supplied sources give no target specific fixation effect (UniProt P35606 subcellular location). | Compare expected High cells and an in-run positive control before calling the section negative; document the compartment actually seen (HPA: tissue IHC; general IHC practice). |
| IF/ICC images seem more reticular than the paraffin IHC result (HPA: subcellular and tissue IHC). | HPA ICC-IF reports mainly endoplasmic reticulum with additional Golgi localization, while HPA tissue IHC describes general cytoplasmic expression; these are distinct observations (HPA: subcellular and tissue IHC). | Interpret the paraffin section against its IHC cell type pattern and controls; use the ICC-IF localization as context rather than an IHC acceptance rule (HPA: tissue IHC and subcellular; general IHC practice). |
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
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use adipose tissue as the negative comparator (HPA: Not detected in adipocytes); on the colon slide, assess neighboring nonglandular cells as internal background comparators 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 control and a concentration-matched rabbit polyclonal isotype control (selected caption: rabbit pAb), plus a COPB2-knockout sample or a validated peptide-block control. For chromogenic colon IHC, quench endogenous peroxidase and check for endogenous biotin if using avidin–biotin detection; inflammatory cells can contribute peroxidase background (standard IHC practice).
⚠️Feasibility: A COPB2-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected paraffin-section caption does not state the fixative (selected caption: fixative unreported). Retrieval dependence is unestablished, although the selected caption uses high-pressure retrieval in 10 mM citrate, pH 6.0, before IHC (selected caption). Frozen sections or IF cannot be judged easier from these data; for colon IHC, distinguish glandular staining from mucin-associated background (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
Troubleshoot COPB2 chromogenic IHC in paraffin sections using the catalog antibody’s reported staining conditions and compartment-aware controls.
How should I adjust retrieval when COPB2 staining is weak in paraffin sections?
Start with heat-mediated retrieval in 10 mM citrate, pH 6.0 (datasheet A07493). The selected paraffin-section image used high-pressure citrate retrieval before staining at 1:50 (caption A07493). If signal is weak, compare short and longer retrieval runs on adjacent sections while keeping antibody dilution and detection constant (standard IHC practice). Include a known-positive glandular control and inspect morphology, because excess heating can damage sections and complicate interpretation (HPA: high in colon glandular cells; standard IHC practice). Record retrieval time and pressure for each run so changes in cytoplasmic staining can be assessed consistently (standard IHC practice).
Could fixation explain inconsistent COPB2 staining between my paraffin blocks?
Target-specific COPB2 sensitivity to fixative type or duration is unknown from the supplied evidence (caption A07493: fixative unstated). Record each block’s fixative and fixation time, then compare adjacent sections processed with the same retrieval and detection conditions (standard IHC practice). Start those comparisons with 10 mM citrate, pH 6.0 retrieval and the reported 1:50 antibody dilution (datasheet A07493; caption A07493). Include a glandular positive control in each run to distinguish a block-level staining problem from a run-level problem (HPA: high in colon glandular cells; standard IHC practice). Interpret weak staining alongside preserved morphology and the control result, without assigning a COPB2-specific fixation effect (standard IHC practice).
Where should COPB2 staining appear, and when is a nuclear pattern suspicious?
Assess COPB2 primarily as cytoplasmic staining, with possible perinuclear membrane-associated accentuation (HPA: general cytoplasmic expression; UniProt P35606: cytosol and Golgi membrane). Coatomer can be cytosolic or assembled on the cytoplasmic face of Golgi-derived vesicles, so a solely crisp nuclear pattern is discordant (UniProt P35606: coatomer localisation). HPA additionally reports endoplasmic reticulum and Golgi localisation by ICC/IF, which can inform compartment checks but does not establish a paraffin-section pattern (HPA: approved subcellular locations). Compare stained sections with a no-primary control and inspect cytoplasm at high magnification (standard IHC practice). Score any nuclear signal separately from cytoplasmic signal rather than combining them into one COPB2 result (standard IHC practice).
Can this stain distinguish COPB2 isoforms or modified epitopes?
Do not assign an isoform from this stain: COPB2 has 2 listed isoforms, while the supplied antibody evidence does not specify an isoform-selective epitope (UniProt P35606: isoforms; caption A07493). The protein has no annotated transmembrane segment, so staining interpretation should follow its cytosolic and membrane-associated locations rather than presume a luminal epitope (UniProt P35606: topology and localisation). Acetyllysine 627 and phosphoresidues 859 and 861 are annotated, but their effect on this antibody’s binding is unreported (UniProt P35606: modified residues; caption A07493). For an isoform or modification claim, obtain epitope mapping and validate the distinction with appropriate reference material (standard IHC practice). Until then, report total detectable COPB2 immunoreactivity under the stated retrieval conditions (standard IHC practice).
How can I check COPB2 localisation by IF alongside the chromogenic IHC result?
Treat IF/ICC as a separate assay and validate the antibody’s fluorescence performance before comparing it with paraffin-section IHC (standard IF practice). Multiplex COPB2 with a marker identifying the cell population under study, then compare intracellular patterns within marker-positive cells (standard IF practice). Choose a fluorophore channel with low tissue autofluorescence and include unstained and single-label controls to check bleed-through (standard IF practice). Because COPB2 is cytosolic and associates with the cytoplasmic face of Golgi and vesicle membranes, use permeabilisation appropriate for access to that compartment (UniProt P35606: topology and localisation; standard IF practice). HPA reports approved endoplasmic reticulum and additional Golgi localisation by ICC/IF; assess agreement without assuming identical patterns across assay formats (HPA: subcellular localisation; standard IF practice).
What should I check when COPB2 IHC shows diffuse brown background?
First compare the stained section with a no-primary control and examine whether brown signal follows tissue structures or appears in debris and cut edges (standard IHC practice). Verify the general chromogenic workflow’s peroxidase block, washing and DAB development; these steps are not COPB2-specific evidence (standard IHC practice). If background persists, titrate from the reported 1:50 dilution and shorten DAB development while retaining the same positive control (caption A07493; standard IHC practice). COPB2 has broad cytoplasmic expression, so diffuse cellular staining alone does not prove nonspecific binding (HPA: general cytoplasmic expression). Judge the result by compartment, cell morphology and control staining together, particularly where damaged tissue traps reagent (UniProt P35606: localisation; standard IHC practice).
How should I quantify COPB2 IHC across specimens with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic compartment before scoring, because COPB2 staining varies across annotated tissue cell types (HPA: tissue IHC profile; standard IHC practice). An H-score can combine the percentage of cells at each intensity into a 0–300 result; alternatively report the percentage of cytoplasm-positive cells using a fixed threshold (standard IHC practice). Normalise counts to the number of eligible cells, or report positive-cell density per mm² of viable tissue when cell counts cannot be obtained (standard IHC practice). Exclude necrotic and edge-damaged regions by a prespecified rule, and keep retrieval, imaging and thresholds consistent (standard IHC practice). Report cell-type composition and control performance alongside scores so differences are not mistaken for altered COPB2 expression (standard IHC practice).
How do I distinguish plausible COPB2 positivity from staining artefact?
A plausible result shows cytoplasmic signal in intact cells, potentially with perinuclear accentuation, consistent with coatomer’s cytosolic and Golgi-associated locations (HPA: general cytoplasmic expression; UniProt P35606: localisation). High staining in colon glandular cells offers a documented positive reference, whereas adipocytes were reported as not detected (HPA: tissue IHC profile). Treat isolated nuclear staining, signal confined to tissue edges or necrosis, and staining reproduced without primary antibody as suspect (UniProt P35606: localisation; standard IHC practice). Check whether endogenous peroxidase contributes to brown signal using the no-primary control and the general peroxidase block (standard IHC practice). Interpret intensity cautiously because HPA rates the tissue IHC profile Approved with medium agreement between staining and RNA expression (HPA: reliability).
Boster reagents
Best COPB2 / Coatomer subunit beta' IHC Antibodies
The catalog antibody A07493 has real IHC data from paraffin-embedded human breast cancer (IHC image caption); IF/ICC is listed for human, mouse and rat (catalog applications and reactivity).
Real IHC data
Anti-Coatomer subunit beta' COPB2 Antibody
Cat # A07493
A07493 will render with an IHC figure from paraffin-embedded human breast cancer stained at 1:50 after citrate pH 6.0 retrieval (IHC image caption). The same SKU lists IF/ICC and human, mouse and rat reactivity, but provides no IF figure (catalog applications, reactivity and image records).
Which to pick: Choose A07493 for paraffin-section IHC: its own caption documents staining of human breast cancer tissue; the fixative is unreported (IHC image caption). For IF/ICC or mouse and rat samples, A07493 is the listed option because those applications and species appear in its catalog record; the provided figure documents human IHC only (catalog applications, reactivity and image records).
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 Cytoplasmic staining in tissue (HPA tissue IHC); Golgi and vesicle association (UniProt). 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: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07493); Caveat: Antibody staining has medium consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage needs validation (UniProt); Regulation: Cell-level intensity varies (HPA tissue IHC). 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.