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CCNB2 / G2/mitotic-specific cyclin-B2 · IHC design guide
Design Immunohistochemistry for CCNB2
Plan CCNB2 IHC-P around cytoplasmic staining in subsets of tissue cells (HPA tissue IHC). Use bone marrow hematopoietic cells as a positive reference and adipocytes as a negative reference (HPA tissue IHC), then compare staining with consistent fixation and scoring.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CCNB2 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 a subset of tissue cells (HPA tissue IHC) | |
| Staining pattern | Subset-positive cytoplasmic cells across several tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M02040) | |
| Positive control | Appendix+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 fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Subset staining can make bulk tissue appear weak (HPA tissue IHC) | |
| Regulation | G2/mitotic-specific expression (UniProt) | |
| Isoform / epitope | No annotated isoforms, signal peptide or propeptide (UniProt) |
Section 1
Recommended CCNB2 IHC & IF Protocols
Compare the catalog antibody’s EDTA pH 8.0 IHC protocol (datasheet M02040) with four published CCNB2 IHC protocols (PMC8728388; PMC11832750; PMC3545739; PMC8331305).
| Sample | Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet M02040) |
| Fixation | Image fixative and duration unreported (datasheet M02040); 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 M02040); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M02040) |
| Primary antibody | Rabbit monoclonal (clone GIG-3) anti-CCNB2, 1:50 (datasheet M02040) |
| Primary incubation | Overnight at 4 °C (datasheet M02040) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M02040) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CCNB2-positive staining in endocrine cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in a subset of cells in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet M02040). For invasive breast carcinoma, a published alternative used EDTA pH 9 at 97°C for 20 min (PMC3545739).
Section 2
What Is the Expected CCNB2 Staining Pattern?
In paraffin-section IHC, expect CCNB2 staining in the cytoplasm of a subset of cells, rather than uniform staining across a tissue (HPA tissue IHC). Reported high-staining cells include bone-marrow hematopoietic cells, appendix and intestinal endocrine cells, and testis Leydig cells (HPA tissue IHC). HPA rates the tissue pattern Enhanced but notes medium consistency with RNA data (HPA tissue IHC). CCNB2 has no transmembrane segment (UniProt O95067 topology).
| Distinct cytoplasmic staining in some hematopoietic cells in bone marrow or endocrine cells in appendix. | This matches reported high staining in those cell populations and the subset pattern across tissues (HPA tissue IHC). Score the identified cells and compartment; a whole-section average can conceal a restricted positive population (standard IHC practice). |
| Strong nuclear-only or sharply membranous staining, with little cytoplasmic signal. | That differs from the reported cytoplasmic tissue pattern (HPA tissue IHC). Recheck cell identity and controls before interpreting it as CCNB2; tissue IHC alone does not establish every possible cell-cycle-dependent location (UniProt O95067 function; HPA tissue IHC). |
| Prominent staining in adipocytes, cardiomyocytes, or lung alveolar cells. | Those cells are reported as not detected (HPA tissue IHC). Check whether the signal follows tissue structures or appears in reagent controls; cross-reactivity or endogenous chromogenic activity are possible explanations (standard IHC practice). |
| Diffuse color across several compartments, including areas without identifiable positive cells. | This obscures the reported subset-specific cytoplasmic pattern (HPA tissue IHC). Compare a primary-antibody omission control and inspect the counterstain before scoring; diffuse background alone cannot establish CCNB2 expression (standard IHC practice). |
| No signal in bone-marrow hematopoietic cells or testis Leydig cells. | Both are reported high-staining populations, so a blank result warrants a run-level check (HPA tissue IHC). Absence in one section is not proof of target absence: assess tissue preservation, controls, and detection before drawing that conclusion (standard IHC practice). |
💡Expected CCNB2 appearanceCall a result positive when identifiable cells show discrete cytoplasmic staining—potentially strong in reported high-staining populations—while widespread nuclear-only, membranous, or cell-independent color prompts review (HPA tissue IHC; standard IHC practice).
| Cell population and sampling | HPA describes expression in a subset of cells; bone-marrow hematopoietic cells, several intestinal endocrine populations, and testis Leydig cells are reported high, whereas adipocytes are not detected (HPA tissue IHC). Choose and score the relevant cells rather than treating every cell in a named tissue as equivalent. |
| Cell-cycle context | CCNB2 controls the G2/M transition and forms a complex with CDK1 (UniProt O95067 function and subunit). Differences between individual cells can be biologically plausible, but morphology and staining alone cannot assign a cell-cycle phase (standard IHC practice). |
| Evidence strength | HPA lists tissue-IHC reliability as Enhanced while describing medium consistency with RNA expression data; two listed antibodies have Enhanced IHC validation (HPA tissue IHC; HPA antibodies). Use that support for pattern selection without treating every unexpected stain as validated. |
| Compartment evidence across applications | Tissue IHC reports cytoplasmic expression (HPA tissue IHC). ICC-IF reports mainly cytosolic localization with additional Golgi localization (HPA subcellular). The Golgi observation can inform an IF/ICC readout but does not require a resolvable Golgi pattern in chromogenic paraffin sections. |
| Situation | Likely cause | Next action |
|---|---|---|
| A reported high-staining population is blank. | The relevant cells may be absent from the examined area, or the IHC run may have failed; HPA reports high staining in bone-marrow hematopoietic cells and testis Leydig cells (HPA tissue IHC). | Confirm cell identity and tissue quality, then review the run's positive control and detection steps before changing conditions (standard IHC practice). |
| Only nuclear or membrane staining is apparent. | The compartment conflicts with HPA's cytoplasmic tissue-IHC profile (HPA tissue IHC); the observation alone does not identify the cause. | Inspect morphology, controls, and staining distribution. Repeat with an independently validated antibody if the unexpected pattern persists (standard IHC practice; HPA antibodies). |
| Unexpectedly strong staining appears in a reported negative cell type. | HPA reports adipocytes, cardiomyocytes, and alveolar cells as not detected (HPA tissue IHC). Cross-reactivity or endogenous detection activity may contribute (standard IHC practice). | Check cell identity and a primary-antibody omission control; assess endogenous activity according to the chosen chromogenic detection system (standard IHC practice). |
| The section has diffuse background that hides cell boundaries. | Background from blocking, washing, or detection can prevent assessment of the expected subset pattern (standard IHC practice; HPA tissue IHC). | Compare the omission control, check reagent and wash steps, and score only signal localized to identifiable cells after the background is resolved (standard IHC practice). |
| The pattern varies between sections of the same tissue. | The reported expression is restricted to subsets, and HPA notes medium consistency between antibody staining and RNA data (HPA tissue IHC). Sampling may change which relevant cells are present. | Compare the same identified cell population across sections and record its cytoplasmic staining separately from the surrounding tissue (HPA tissue IHC; standard IHC practice). |
| In IF/ICC, where should CCNB2 signal be expected? | HPA reports mainly cytosolic localization, with additional Golgi localization, from ICC-IF evidence (HPA subcellular). | Interpret signal against those compartments and suitable IF/ICC controls; use the separate IF/ICC guide for that application's procedure (HPA subcellular; standard IF practice). |
Experiment evidence
IHC & IF images showing CCNB2 staining
M02040 · IHCIHC analysis of CCNB2 using anti-CCNB2 antibody (M02040). CCNB2 was detected in a paraffin-embedded section of human colon adenocarcinoma ti
M02040 · IHCIHC analysis of CCNB2 using anti-CCNB2 antibody (M02040). CCNB2 was detected in a paraffin-embedded section of human colon adenocarcinoma ti
M02040 · IFImmunofluorescent analysis using the Antibody at 1:50 dilution.
A02040-1 · IHCIHC analysis of CCNB2 using anti-CCNB2 antibody (A02040-1). CCNB2 was detected in paraffin-embedded section of human Ovarian cancer tissue.
A02040-1 · IHCIHC analysis of CCNB2 using anti-CCNB2 antibody (A02040-1). CCNB2 was detected in paraffin-embedded section of human mammary cancer tissue.
A02040-1 · IFIF analysis of CCNB2 using anti-CCNB2 antibody (A02040-1). CCNB2 was detected in immunocytochemical section of U20S cells. Enzyme antigen re
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000157456-CCNB2/tissue/cerebral+cortex#imid_2871356
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000157456-CCNB2/tissue/colon#imid_2871290
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000157456-CCNB2/tissue/liver#imid_2871283
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000157456-CCNB2/tissue/kidney#imid_2871284
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA008873; green = CCNB2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA008873; green = CCNB2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CCNB2 IHC & IF
🧪Run colon first and score its endocrine cells, which HPA rates High for CCNB2 (HPA: colon endocrine cells, High). Use adipose tissue adipocytes as the negative tissue (HPA: adipocytes, Not detected); on the colon slide, treat neighboring cells without specific staining as background references without assuming that every nonendocrine cell is CCNB2-negative (HPA: colon endocrine-cell row only).
Positive control tissue: Appendix (Endocrine cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CCNB2 in A-431, U-251MG, U2OS, with annotated localisation: Cytosol (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a species- and isotype-matched rabbit control IgG, and a CCNB2 knockout control or peptide block where available (catalog caption: rabbit primary; standard IHC practice). For chromogenic colon sections, block endogenous peroxidase and check inflammatory cells for residual signal; block endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: The selected M02040 paraffin-section caption reports heat retrieval in EDTA at pH 8.0, but its fixative is unreported; a target-specific fixation window or fixation effect is also unreported in the supplied evidence (selected-SKU tissue-IHC caption). The caption supports a practical chromogenic paraffin IHC starting point, while the supplied evidence does not establish that frozen sections or IF are easier; HPA does show ICC-IF images in A-431, U-251MG and U2OS cells (selected-SKU tissue-IHC caption; HPA: subcellular images). In colon sections, distinguish endocrine-cell staining from luminal material and residual endogenous peroxidase signal (HPA: colon endocrine cells, High; standard IHC practice).
HPA tissue IHC evidence for CCNB2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Endocrine cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Colon | Endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | High | Protein (IHC) | HPA → |
| Rectum | Endocrine cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lung | Alveolar cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CCNB2 IHC Tips
Troubleshoot CCNB2 staining in paraffin sections by checking retrieval, cell-specific localisation and assay controls before interpreting chromogenic signal.
How should I retrieve CCNB2 antigen when staining paraffin sections?
Start with heat-mediated antigen retrieval in EDTA pH 8.0 (datasheet M02040). The selected paraffin-section example used that retrieval before a 1:50 primary incubation overnight at 4°C; its caption does not report a fixative (datasheet M02040). If signal is weak, compare a modestly adjusted heating duration on matched sections while keeping detection and exposure to DAB consistent, then assess tissue preservation and staining together (standard IHC practice). Include the same positive-control section in each run so a retrieval change is judged against reproducible CCNB2 staining rather than a change in specimen composition (standard IHC practice).
Could fixation explain weak or uneven CCNB2 staining?
Target-specific sensitivity of CCNB2 to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not state its fixative (datasheet M02040). Record each specimen’s fixative and processing history, and compare sections processed together before assigning a weak result to fixation (standard IHC practice). Examine morphology and staining across intact tissue, since damaged or poorly preserved areas can make chromogenic signal difficult to interpret (standard IHC practice). If a processing difference is suspected, repeat the same EDTA pH 8.0 retrieval and detection conditions on matched material, using a concurrently stained positive control to separate assay failure from specimen variation (datasheet M02040; standard IHC practice).
What staining compartment should I expect for CCNB2 in tissue?
Expect staining in a subset of cells, predominantly in the cytoplasm, when evaluating tissue sections (HPA: tissue IHC profile). The subcellular dataset places CCNB2 mainly in the cytosol and additionally at the Golgi apparatus, although those observations come from ICC/IF images rather than the selected paraffin section (HPA: subcellular). Compare signal with cell morphology and a counterstain before calling a focal cytoplasmic pattern positive (standard IHC practice). Diffuse staining across every compartment, especially when accompanied by tissue-edge intensity or poor morphology, deserves a control review; UniProt supplies no subcellular annotation that would independently establish a tissue-specific pattern (UniProt O95067; standard IHC practice).
Could isoforms or modifications change how this antibody stains CCNB2?
The supplied UniProt record lists 0 isoforms and a single chain spanning residues 1–398, so it provides no documented isoform pattern to explain discordant staining (UniProt O95067). It lists phosphorylation at several residues, including Thr8 and Ser11, but the selected caption does not identify the antibody epitope or show modification-dependent staining (UniProt O95067; datasheet M02040). Treat an apparent compartment shift or loss of signal as an assay finding until it is reproduced with controls and independently supported (standard IHC practice). Compare matched sections with consistent retrieval and primary-antibody conditions before attributing any difference to epitope accessibility (standard IHC practice).
How should I cross-check a chromogenic CCNB2 result by IF?
Use IF/ICC as a separate cross-check, pairing CCNB2 with a marker that identifies the cell population under examination and comparing both signals with nuclear morphology (standard IF practice). HPA reports cytosolic CCNB2 with an additional Golgi location in ICC/IF images, so plan permeabilisation for an intracellular epitope while recognising that the antibody’s exact epitope is unspecified (HPA: subcellular; datasheet M02040). Choose fluorophores after inspecting tissue autofluorescence in an unstained control, and favour channels with clear separation from that background (standard IF practice). Compare the IF cell-level pattern with cytoplasmic staining in the chromogenic section without assuming identical sensitivity between methods (HPA: tissue IHC profile; standard IF practice).
What should I check when DAB staining is widespread?
First compare a no-primary control with the test section to identify staining attributable to the detection system or tissue rather than primary-antibody binding (standard IHC practice). The selected example used 10% goat serum blocking, a peroxidase-conjugated anti-rabbit secondary and DAB development after 1:50 primary antibody overnight at 4°C (datasheet M02040). Check peroxidase blocking, washes and chromogen development as general chromogenic IHC steps when background persists (standard IHC practice). Interpret widespread uniform colour cautiously because HPA describes cytoplasmic CCNB2 in only a subset of cells across several tissues (HPA: tissue IHC profile).
How should I score heterogeneous CCNB2 staining in paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue area before scoring, since HPA describes cytoplasmic staining in a subset of cells rather than uniform tissue-wide expression (HPA: tissue IHC profile). Report the percentage of positive cells and staining intensity, or use an H-score when those categories are scored consistently across sections (standard IHC practice). For spatial comparisons, report positive-cell density per mm² of viable, evaluable tissue and state whether analysis excludes necrosis and damaged edges (standard IHC practice). Normalise comparisons to the same cell population and sampled area, and keep retrieval, chromogen development and positivity thresholds consistent across the run (standard IHC practice).
How can I distinguish genuine CCNB2 staining from artefact?
Look for reproducible cytoplasmic signal in identifiable cells, consistent with the tissue IHC profile and the mainly cytosolic ICC/IF localisation (HPA: tissue IHC profile; HPA: subcellular). HPA reports high staining in bone-marrow hematopoietic cells and testicular Leydig cells, but these patterns do not establish that every positive cell is in mitosis (HPA: tissue IHC; UniProt O95067 function). Treat staining limited to cut edges, necrotic regions or a no-primary control as suspect, and review endogenous peroxidase blocking if DAB appears without primary antibody (standard IHC practice). Correlate a proposed G2/M interpretation with cell morphology or an independently assessed cell-cycle marker, because CCNB2 participates in G2/M control (UniProt O95067 function; standard IHC practice).
Boster reagents
Best CCNB2 / G2/mitotic-specific cyclin-B2 IHC Antibodies
Both catalog antibodies have human paraffin-section IHC images; A02040-1 also has a cell-based IF image (catalog image captions). Both list human, mouse, and rat reactivity (catalog applications/reactivity).
Real IHC data
Anti-Cyclin B2 CCNB2 Rabbit Monoclonal Antibody
Cat # M02040
Real IHC data
Anti-Cyclin B2/CCNB2 Antibody ®
Cat # A02040-1
M02040 will render with a human colon adenocarcinoma paraffin-section IHC figure and lists IHC and IF/ICC applications (M02040 image caption; catalog applications). A02040-1 will render with a human ovarian cancer paraffin-section IHC figure and also has a cell-based IF image (A02040-1 image captions).
Which to pick: For tissue IHC, choose M02040 for a rabbit monoclonal tested at 1:50 on a human paraffin section, or A02040-1 for a rabbit antibody tested at 1 μg/ml on a human paraffin section (catalog antibody details; respective IHC image captions). For IF/ICC, A02040-1 has a cell-based IF image at 2 μg/ml; M02040 lists IF/ICC and has IF images with limited sample details (catalog applications; respective IF image captions). Both list human, mouse, and rat reactivity, but the supplied IHC captions show human tissue only; neither IHC caption reports the fixative (catalog reactivity; respective IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O95067 (CCNB2_HUMAN, G2/mitotic-specific cyclin-B2).
- Human Protein Atlas. CCNB2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CCNB2 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the Golgi apparatus..
- Human Protein Atlas. CCNB2 antibody validation summary (2 antibodies).
- Prognostic Significance of CCNB2 Expression in Triple-Negative Breast Cancer. Cancer management and research 2021 — PMC8728388.
- Knockdown of CCNB2 inhibits the tumorigenesis of gastric cancer by regulation of the PI3K/Akt pathway. Scientific reports 2025 — PMC11832750.
- Elevated cyclin B2 expression in invasive breast carcinoma is associated with unfavorable clinical outcome. BMC cancer 2013 — PMC3545739.
- Cyclin B2 (CCNB2) Stimulates the Proliferation of Triple-Negative Breast Cancer (TNBC) Cells In Vitro and In Vivo. Disease markers 2021 — PMC8331305.
- PubMed PMID:11230166 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CCNB2 (G2/mitotic-specific cyclin-B2). Expected localisation Cytoplasmic staining in a subset of tissue cells (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M02040). Validated IHC antibody: M02040 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Endocrine cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet M02040); 2. Fixation: Image fixative and duration unreported (datasheet M02040); 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 M02040); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M02040); 8. Primary antibody: Rabbit monoclonal (clone GIG-3) anti-CCNB2, 1:50 (datasheet M02040); 9. Primary incubation: Overnight at 4 °C (datasheet M02040). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Subset staining can make bulk tissue appear weak (HPA tissue IHC); Isoform / epitope: No annotated isoforms, signal peptide or propeptide (UniProt); Regulation: G2/mitotic-specific expression (UniProt). Evidence: UniProt O95067, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the Golgi…, Datasheet M02040, PMC8728388, PMC11832750. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CCNB2/immunohistochemistry-ihc-guide.