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CSNK2B / Casein kinase II subunit beta · IHC design guide
Design Immunohistochemistry for CSNK2B
Plan CSNK2B staining in paraffin sections using the IHC-validated antibody at 2–5 μg/ml (datasheet A03475-1). Compare nuclear and cytoplasmic staining with the reported tissue pattern, while accounting for the antibody specificity caution (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CSNK2B 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 | Nuclear and cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Glandular, glial and other cells show nuclear and cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03475-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining may detect proteins from more than one gene (HPA tissue IHC) | |
| Regulation | Expression regulation is not annotated (UniProt) | |
| Isoform / epitope | No isoforms annotated; chain spans residues 2–215 (UniProt) |
Section 1
Recommended CSNK2B IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 heat retrieval (datasheet A03475-1). The published IHC protocols below stain colorectal tissue microarrays (PMC8222461) and paraffin sections (PMC7210266).
| Sample | Paraffin-embedded human gallbladder adenocarcinoma tissue; fixative not specified (datasheet A03475-1) |
| Fixation | Image fixative and duration unreported (datasheet A03475-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A03475-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03475-1) |
| Primary antibody | Rabbit anti-CSNK2B, 2-5 μg/ml (datasheet A03475-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03475-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03475-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CSNK2B-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic and nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval (datasheet A03475-1). Compare citrate pH 6.0 (PMC8222461) or CC1 pH 8.4 (PMC7210266) when adapting those protocols.
Section 2
What Is the Expected CSNK2B Staining Pattern?
CSNK2B is nuclear (UniProt P67870), while tissue IHC shows general nuclear and cytoplasmic staining (HPA: tissue IHC). Expect staining in adrenal glandular cells, caudate glia and cerebellar molecular-layer cells, each reported High (HPA: tissue IHC). Interpret intensity with caution: the tissue profile is Approved with medium RNA–staining consistency, and HPA warns that the antibodies target protein from more than one gene (HPA: tissue IHC). CSNK2B has no transmembrane segment (UniProt P67870 topology).
| Nuclear staining, with some cytoplasmic staining, in expected cells. | This fits the reported tissue pattern (HPA: tissue IHC) and nuclear assignment (UniProt P67870). Compare cells within the same section and a known-positive section before scoring intensity (general IHC practice). |
| Signal is confined to cell membranes, with no convincing nuclear staining. | A membrane-only pattern does not fit the nuclear assignment or reported nuclear/cytoplasmic tissue pattern (UniProt P67870; HPA: tissue IHC). Check staining controls and localization before treating it as CSNK2B. |
| Strong staining appears in cells where the chosen tissue profile reports Low staining. | Consider off-target staining or endogenous detection activity (general IHC practice). Low is not negative, and HPA reports no negative tissue in this profile (HPA: tissue IHC); intensity alone cannot establish cross-reactivity. |
| Diffuse chromogen covers cells and surrounding tissue, obscuring compartments. | This is uninterpretable background rather than a scorable localization pattern (general IHC practice). Review blocking, detection and wash controls before comparing it with HPA's nuclear/cytoplasmic profile (HPA: tissue IHC). |
| No signal is visible in a section expected to stain strongly. | Adrenal glandular cells or caudate glia provide reported High examples (HPA: tissue IHC). First assess the assay and section controls (general IHC practice); one blank section does not establish absent CSNK2B expression. |
💡Expected CSNK2B appearanceCall a positive when expected cells show interpretable nuclear staining, with possible cytoplasmic staining and relatively strong signal in HPA High examples; isolated membrane staining or diffuse chromogen is suspect (UniProt P67870; HPA: tissue IHC; general IHC practice).
| Tissue and cell choice | HPA reports High staining in adrenal glandular cells, caudate glia and cerebellar molecular-layer cells, but Low staining in several other listed cell types (HPA: tissue IHC). Low examples are not validated negatives. |
| Strength of tissue evidence | The tissue profile is Approved, with medium RNA–staining consistency and a warning that antibodies target protein from more than one gene (HPA: tissue IHC). Treat a plausible pattern as supportive, not definitive identification. |
| Antibody validation | Four listed antibodies have Approved IHC status (HPA: antibodies). That status supports use of their reported IHC patterns; it does not establish the specificity of a different catalog antibody or a particular section. |
| IF/ICC Q&A: Where should signal appear? | Mainly in the nucleoplasm; HPA also reports supported nucleolar fibrillar-center localization and approved additional locations in sperm structures (HPA: subcellular ICC-IF). Assess IF/ICC on its own guide page. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive cells appear blank. | The IHC run or section may have failed; absence alone cannot distinguish assay failure from a biological difference (general IHC practice). | Check detection and section controls, then repeat with a reported High example such as adrenal glandular cells (HPA: tissue IHC; general IHC practice). |
| Staining is cytoplasmic with little nuclear signal. | Cytoplasmic tissue staining is reported, but a nuclear component is expected from the localization evidence (HPA: tissue IHC; UniProt P67870). | Review nuclear counterstain alignment and a known-positive section; avoid calling a cytoplasmic-only result definitive (general IHC practice). |
| A crisp membrane rim dominates the image. | A membrane-only pattern conflicts with reported localization and the lack of a transmembrane segment (HPA: tissue IHC; UniProt P67870 topology). | Inspect detection controls and compare cellular compartments in a reported High tissue before scoring the rim (general IHC practice; HPA: tissue IHC). |
| Low-profile cells stain as strongly as expected High cells. | HPA Low means some reported staining, and its tissue profile carries a multi-gene antibody warning (HPA: tissue IHC). | Compare cell types on matched sections and review controls; do not use a Low example as a negative control (general IHC practice; HPA: tissue IHC). |
| Chromogen is diffuse or covers tissue outside cells. | Background can obscure the nuclear/cytoplasmic pattern (general IHC practice; HPA: tissue IHC). | Check blocking, washes and detection controls, then score only cells whose boundaries and compartments remain interpretable (general IHC practice). |
| IF/ICC localization seems inconsistent with the IHC slide. | HPA's ICC-IF profile is mainly nucleoplasmic, while its tissue IHC profile reports both nuclear and cytoplasmic expression (HPA: subcellular ICC-IF; tissue IHC). | Compare each image with its own assay-specific reference and controls; do not transfer an IF/ICC localization call directly into an IHC score (general IHC practice). |
Experiment evidence
IHC & IF images showing CSNK2B staining
A03475-1 · IHCIHC analysis of CSNK2B using anti-CSNK2B antibody (A03475-1). CSNK2B was detected in a paraffin-embedded section of human gallbladder adenoc
A03475-1 · IHCIHC analysis of CSNK2B using anti-CSNK2B antibody (A03475-1). CSNK2B was detected in a paraffin-embedded section of human liver cancer tissu
M03475 · IHCFigure 2. IHC analysis of CSNK2B using anti-CSNK2B antibody (M03475). CSNK2B was detected in a paraffin-embedded section of human colon tiss
M03475 · IHCFigure 3. IHC analysis of CSNK2B using anti-CSNK2B antibody (M03475). CSNK2B was detected in a paraffin-embedded section of human colon canc
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000204435-CSNK2B/tissue/cerebral+cortex#imid_1624980
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000204435-CSNK2B/tissue/colon#imid_1624922
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000204435-CSNK2B/tissue/liver#imid_1624915
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000204435-CSNK2B/tissue/kidney#imid_1624916
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB013087; green = CSNK2B, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB013087; green = CSNK2B, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CSNK2B IHC & IF
🧪Run adrenal gland first and look for staining in its glandular cells (HPA: adrenal gland glandular cells High). HPA lists no negative tissue because CSNK2B is detected in all 45 scored tissues, so no-primary and isotype controls carry the negative comparison; cells without specific staining on the positive slide should show only background, but no cell population there is established as a biological negative (HPA: no negative rows; detected in all 45 scored tissues).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: CSNK2B is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CSNK2B in A-431, U-251MG, U2OS, CACO-2, MCF-7, Sperm, KOLF2.1J, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a control using nonimmune rabbit IgG matched to the primary antibody’s isotype and concentration (caption: rabbit anti-CSNK2B); use CSNK2B knockout tissue or a validated peptide block as a biological specificity control. For adrenal gland sections, block endogenous peroxidase and assess endogenous biotin before interpreting SABC/DAB staining (caption: SABC/DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A03475-1 paraffin-section caption does not state its fixative (caption: fixative not stated). The reported IHC procedure used heat-mediated retrieval in EDTA at pH 8.0, but the evidence does not establish whether retrieval is required under other conditions (caption: EDTA retrieval, pH 8.0). The supplied evidence does not establish frozen sections or IF as easier; for adrenal gland IHC, endogenous peroxidase or biotin may complicate SABC/DAB interpretation (caption: SABC/DAB detection; HPA: adrenal gland glandular cells High).
HPA tissue IHC evidence for CSNK2B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Breast | Myoepithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: CSNK2B is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced CSNK2B IHC Tips
Troubleshoot CSNK2B staining in paraffin sections by checking retrieval, compartment, cell type and detection controls together.
How should I troubleshoot weak CSNK2B staining after antigen retrieval?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A03475-1). The selected tissue image used this retrieval before incubation with 2 μg/ml antibody overnight at 4°C (datasheet A03475-1). If staining is weak, compare slides processed with the same heating and cooling conditions, then check antibody incubation and detection before changing retrieval (standard IHC practice). The caption gives no heating duration, so establish a reproducible time for your equipment and assess tissue preservation alongside staining (datasheet A03475-1; standard IHC practice).
Does fixation change CSNK2B staining in paraffin sections?
Target-specific fixation sensitivity is unknown from the supplied evidence; the selected image describes paraffin-embedded tissue but does not state its fixative (datasheet A03475-1). Do not interpret that image as evidence for a particular fixation method or duration (datasheet A03475-1). For a controlled comparison, process matched specimens consistently, record fixation conditions, and apply the same EDTA pH 8.0 retrieval and detection workflow to every section (datasheet A03475-1; standard IHC practice). If staining differs, inspect morphology and a positive control before attributing the change to CSNK2B epitope sensitivity (standard IHC practice).
Should CSNK2B stain nuclei, cytoplasm, or both?
Assess nuclear and cytoplasmic staining separately: UniProt lists CSNK2B in the nucleus, while tissue IHC reports general cytoplasmic and nuclear expression (UniProt P67870 subcellular; HPA tissue IHC). Nucleoplasmic localisation is supported by subcellular imaging, but that observation does not require every paraffin section to show exclusively nuclear chromogen (HPA subcellular; HPA tissue IHC). Compare each compartment with the same tissue's morphology and no-primary control before scoring (standard IHC practice). Treat an unfamiliar pattern cautiously because the tissue IHC antibody carries a warning that it targets protein from more than one gene (HPA tissue IHC).
Could epitope choice explain inconsistent CSNK2B staining?
The record lists 0 annotated isoforms and several modified residues, including phosphorylation at positions 2, 3, 8 and 37 (UniProt P67870). No epitope map or evidence that these modifications change staining is supplied, so neither can explain a discrepant section by itself (supplied evidence). Keep EDTA pH 8.0 retrieval and antibody incubation consistent while comparing sections (datasheet A03475-1; standard IHC practice). If an unexpected pattern persists, check it with an independent specificity control and report which antibody produced each result (standard IHC practice).
How can IF help assess a disputed IHC pattern?
For a separate IF assessment, multiplex CSNK2B with a validated marker for the cell population under examination; tissue IHC reports high staining in adrenal glandular cells and caudate glial cells (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and consider a far-red channel when tissue autofluorescence obscures shorter wavelengths (standard IF practice). Permeabilise cells or sections for access to intracellular nuclear and cytoplasmic epitopes; CSNK2B has no transmembrane segment (UniProt P67870 topology; HPA tissue IHC; standard IF practice). Include single-stain and no-primary controls, and validate the antibody for IF before using fluorescence to adjudicate chromogenic IHC (standard IF practice).
What causes diffuse brown background in CSNK2B IHC?
The selected image used a biotinylated secondary, streptavidin–biotin detection and DAB chromogen (datasheet A03475-1). Check a no-primary section for signal from the detection system, including endogenous biotin or peroxidase activity; apply a peroxidase block as a general chromogenic IHC step (standard IHC practice). Its section was blocked with 10% goat serum, and the secondary incubation was 30 minutes at 37°C (datasheet A03475-1). If background persists, compare blocked controls and reagent exposure across sections before interpreting diffuse brown deposits as cytoplasmic CSNK2B (standard IHC practice; HPA tissue IHC).
How should I quantify CSNK2B staining across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score nuclear and cytoplasmic compartments separately because both occur in tissue IHC (HPA tissue IHC; standard IHC practice). For each compartment, record the percentage of positive cells and an H-score using intensity grades 0–3 multiplied by the percentage at each grade, yielding 0–300 (standard IHC scoring practice). Normalise cell counts to all eligible cells of the same type, or report positive-cell density per mm² of evaluable tissue (standard IHC scoring practice). Exclude damaged areas consistently and keep retrieval, detection and scoring thresholds fixed across the comparison (standard IHC practice).
How can I distinguish genuine CSNK2B staining from artefact?
Look for reproducible staining in intact cells with an interpretable nuclear or cytoplasmic pattern, rather than isolated deposits at section edges or necrotic areas (HPA tissue IHC; standard IHC practice). High staining is reported in adrenal glandular cells and caudate glial cells, while bronchial respiratory epithelial cells are reported low, not negative (HPA tissue IHC). Review a no-primary control for endogenous enzyme or detection-system signal, particularly with the selected DAB and biotin workflow (datasheet A03475-1; standard IHC practice). Confirm consequential findings independently because the HPA tissue antibody has medium RNA–staining consistency and a warning about recognition of more than one gene (HPA tissue IHC).
Boster reagents
Best CSNK2B / Casein kinase II subunit beta IHC Antibodies
Both antibodies have real IHC images from human paraffin sections; M03475 also has a rat testis image (catalog captions). Neither lists IF/ICC (catalog applications).
Real IHC data
Anti-CSNK2B Antibody ®
Cat # A03475-1
Real IHC data
Anti-Casein Kinase 2 beta Monoclonal Antibody
Cat # M03475
A03475-1 will render with its human gallbladder adenocarcinoma IHC image; its additional captions show human liver, lung, and thyroid cancer sections (A03475-1 IHC captions). M03475 will render with its human colon IHC image; additional captions show human colon and stomach cancer and rat testis sections (M03475 IHC captions).
Which to pick: For paraffin-section IHC, choose A03475-1 for a 2–5 μg/ml starting range (A03475-1 catalog IHC dilution), or M03475 for a rabbit monoclonal at 1:50 (M03475 catalog host, clone, and IHC dilution). Both pictured protocols use EDTA retrieval at pH 8.0; neither caption reports the fixative (A03475-1 and M03475 IHC captions). Neither is listed for IF/ICC; for cross-species IHC, M03475 has a rat tissue image, while mouse appears only in the listed reactivity for both antibodies (catalog applications, reactivity, and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P67870 (CSK2B_HUMAN, Casein kinase II subunit beta).
- Human Protein Atlas. CSNK2B tissue IHC expression (reliability: Approved).
- Human Protein Atlas. CSNK2B subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nucleoli fibrillar center, perinuclear theca, calyx and mid piece..
- Human Protein Atlas. CSNK2B antibody validation summary (5 antibodies).
- CSNK2B contributes to colorectal cancer cell proliferation by activating the mTOR signaling. Journal of cell communication and signaling 2021 — PMC8222461.
- Overexpression of NELFE contributes to gastric cancer progression via Wnt/β-catenin signaling-mediated activation of CSNK2B expression. Journal of experimental & clinical cancer research : CR 2021 — PMC7851912.
- A RUNX2 stabilization pathway mediates physiologic and pathologic bone formation. Nature communications 2020 — PMC7210266.
- Tumor necrosis factor α-induced protein 1 as a novel tumor suppressor through selective downregulation of CSNK2B blocks nuclear factor-κB activation in hepatocellular carcinoma. EBioMedicine 2020 — PMC6950786.
- PubMed PMID:2666134 — UniProt-cited evidence.
- PubMed PMID:1694965 — UniProt-cited evidence.
- PubMed PMID:2513884 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CSNK2B (Casein kinase II subunit beta). Expected localisation Nuclear and cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03475-1). Validated IHC antibody: A03475-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human gallbladder adenocarcinoma tissue; fixative not specified (datasheet A03475-1); 2. Fixation: Image fixative and duration unreported (datasheet A03475-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A03475-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03475-1); 8. Primary antibody: Rabbit anti-CSNK2B, 2-5 μg/ml (datasheet A03475-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03475-1). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining may detect proteins from more than one gene (HPA tissue IHC); Isoform / epitope: No isoforms annotated; chain spans residues 2–215 (UniProt); Regulation: Expression regulation is not annotated (UniProt). Evidence: UniProt P67870, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A03475-1, PMC8222461, PMC7210266. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CSNK2B/immunohistochemistry-ihc-guide.