This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
CKS2 / Cyclin-dependent kinases regulatory subunit 2 · IHC design guide
Design Immunohistochemistry for CKS2
Plan chromogenic paraffin-section CKS2 IHC with bone marrow hematopoietic cells as a high-staining reference (HPA tissue IHC) and a catalog antibody starting range of 1:100–1:300 (datasheet). Interpret nuclear staining with the reported cross-gene specificity caution in mind (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CKS2 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 staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Nuclear staining in most tissues; high in marrow hematopoietic cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adipose tissue+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 paraffin-section fixation consistent (standard IHC practice; not target-specific) | |
| Caveat | Tissue IHC may detect protein from more than one gene (HPA tissue IHC) | |
| Regulation | Tissue-enhanced RNA: bone marrow, testis (HPA tissue RNA) | |
| Isoform / epitope | 0 annotated isoforms; one 1–79 chain (UniProt) |
Section 1
Recommended CKS2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published CKS2 chromogenic IHC methods for lung adenocarcinoma, lower grade glioma, and cervical cancer (PMC9406629; PMC8558665; PMC8806895; PMC6411633).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A08202-1) |
| Fixation | Image fixative and duration unreported (datasheet A08202-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| 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-CKS2, 1:100 - 1:300 (datasheet A08202-1) |
| 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 | CKS2-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen-retrieval rule). The glioma study reports pressure-cooker retrieval in citrate pH 6.0 for 5 min (PMC8806895).
Section 2
What Is the Expected CKS2 Staining Pattern?
In paraffin-section IHC, expect predominantly nuclear CKS2 staining in most tissues (HPA tissue IHC: nuclear expression in most tissues). HPA reports high staining in bone marrow hematopoietic cells, bronchial respiratory epithelial cells and adipocytes, among other listed cells (HPA tissue IHC: High). The target has no annotated transmembrane segment (UniProt P33552 topology). Interpret this pattern cautiously: HPA rates tissue IHC Approved but warns that its antibodies target proteins from more than one gene (HPA tissue IHC: reliability description).
| Nuclear chromogen is strongest in listed high-staining cell populations. | This fits the reported tissue pattern (HPA tissue IHC: nuclear expression; High in bone marrow hematopoietic cells). Score the named cells and their compartment separately; a high tissue entry does not mean every cell in that section should stain. |
| Cytoplasmic or punctate signal dominates an IHC section, with little nuclear staining. | This differs from the tissue IHC profile (HPA tissue IHC: nuclear expression in most tissues). Check morphology, counterstain and detection controls before calling it CKS2. Cytosolic and mitochondrial staining is reported for ICC-IF, a separate assay (HPA subcellular: approved locations). |
| Strong chromogen appears mainly in cells outside the listed high-staining populations. | Treat this as uncertain staining, rather than assigning those cells a validated CKS2 pattern. Cross-reactivity is a concern because the tissue profile carries a multiple-gene warning (HPA tissue IHC: reliability description); endogenous detection activity is another general chromogenic IHC possibility. |
| Color covers much of the section without clear cell or nuclear boundaries. | Diffuse background prevents a compartment call. Inspect the no-primary control and tissue morphology, then review blocking, washes and detection conditions (general IHC practice). The HPA nuclear profile cannot validate an indistinct deposit (HPA tissue IHC: profile). |
| No stain appears in a section containing bone marrow hematopoietic cells. | This conflicts with a reported high-staining population (HPA tissue IHC: High in bone marrow hematopoietic cells), but one blank section does not establish CKS2 absence. Check that the expected cells are present, then compare a reference section and detection controls. |
💡Expected CKS2 appearanceCall a positive result when discernible nuclei stain in the relevant cells, with strong signal in an HPA-listed high population (HPA tissue IHC: nuclear profile; High); widespread featureless color or signal confined to unexpected cells is suspect.
| Tissue and cell selection | Use a documented high-staining population, such as bone marrow hematopoietic cells or bronchial respiratory epithelial cells, as an interpretation reference (HPA tissue IHC: High). HPA supplies no negative or low-staining entries in this payload, so do not designate one as a CKS2-negative tissue. |
| Assay-dependent localisation | Tissue IHC is predominantly nuclear (HPA tissue IHC: profile); ICC-IF mainly shows cytosol and mitochondria, with additional nucleoplasm and vesicles (HPA subcellular: approved locations). Its images come from A-549, PC-3 and U2OS (HPA subcellular: cell lines); those observations do not define a paraffin-IHC protocol. |
| Antibody validation | The tissue IHC record is Approved yet pending external verification because antibodies target proteins from more than one gene (HPA tissue IHC: reliability description). HPA030762 has IHC Approved status; HPA003424 has ICC Approved status (HPA antibodies). Interpret agreement with the pattern as supportive, rather than proof of molecular specificity. |
| Target structure and processing | CKS2 is annotated as one chain spanning residues 1–79, with no signal peptide, propeptide, annotated isoforms or transmembrane segment (UniProt P33552). These annotations give no basis here for a shed extracellular pool or an isoform-specific staining call; the epitope location is not supplied. |
| RNA versus protein evidence | Bone marrow and testis are tissue-enhanced at the RNA level (HPA tissue IHC: RNA specificity). Bone marrow hematopoietic cells also have a High protein-staining entry (HPA tissue IHC: High); the supplied entries do not establish a corresponding high protein-staining call for testis. |
| Situation | Likely cause | Next action |
|---|---|---|
| A documented high-staining population is blank. | The expected cells may be absent from the section, or an IHC preparation or detection step may have failed (general IHC practice). | Confirm cell identity on the counterstain; check the IHC-validated antibody, retrieval and detection workflow against its instructions, then compare a reference section (HPA tissue IHC: High populations). |
| Nuclear detail is lost under diffuse chromogen. | Background or overdevelopment can obscure localisation (general chromogenic IHC practice). | Inspect a no-primary control; review blocking, washing and development conditions. Score localisation only where nuclei and cell outlines remain readable (general IHC practice). |
| Signal is mainly cytoplasmic in paraffin IHC. | The observed compartment differs from the reported tissue IHC profile (HPA tissue IHC: nuclear expression in most tissues). | Recheck compartment assignment using the counterstain and controls. Do not transfer the ICC-IF cytosol and mitochondria call directly to IHC (HPA subcellular: approved locations). |
| Unexpected cells stain strongly. | Antibody cross-reactivity is plausible under HPA's multiple-gene caution (HPA tissue IHC: reliability description); endogenous detection activity is a general chromogenic IHC alternative. | Compare tissue morphology and a no-primary control; review detection blocking and seek independent specificity evidence before identifying the signal as CKS2. |
| A proposed negative tissue also stains. | No negative tissue is documented in the supplied HPA tissue IHC entries (HPA tissue IHC: negative list empty). | Do not use that tissue as a validated negative benchmark. Use reagent controls for background assessment and keep the tissue's CKS2 status unresolved. |
| An IF image appears inconsistent with nuclear IHC. | The assays and reported locations differ (HPA tissue IHC: nuclear profile; HPA subcellular: mainly cytosol and mitochondria), and HPA lists different approved antibodies for IHC and ICC (HPA antibodies). | Interpret each assay against its own HPA record; assess antibody specificity before treating the difference as a biological redistribution. |
Experiment evidence
IHC & IF images showing CKS2 staining
A08202-1 · IHCImmunohistochemistry analysis of paraffin-embedded human brain tissue, using CKS2 Antibody. The picture on the right is blocked with the syn
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000123975-CKS2/tissue/cerebral+cortex#imid_8720380
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000123975-CKS2/tissue/colon#imid_8712370
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000123975-CKS2/tissue/liver#imid_8712371
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000123975-CKS2/tissue/kidney#imid_8712372
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA003424; green = CKS2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA003424; green = CKS2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CKS2 IHC & IF
🧪Run appendix first and assess staining in glandular cells (HPA: High in appendix glandular cells). HPA detects CKS2 in all 45 scored tissues, so no negative tissue is supported; use no-primary and isotype controls, and treat unstained cells on the appendix section as background references rather than validated CKS2-negative cells (HPA: all 45 tissues detected).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: CKS2 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 CKS2 in A-549, PC-3, U2OS, with annotated localisation: Mitochondria (approved), Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section and an irrelevant antibody matched to the primary antibody’s host species, immunoglobulin class, and clonality; test specificity with a CKS2 knockout sample if available or a synthesized-peptide block (selected-SKU tissue-IHC caption: peptide-blocked image). Quench endogenous peroxidase before chromogenic detection, especially in leukocyte-rich areas of appendix (standard IHC practice).
⚠️Feasibility: A CKS2-specific fixation window and antigen-retrieval dependence are unreported in the supplied evidence (supplied target/application evidence). The A08202-1 paraffin-section brain caption leaves the fixative unreported (selected-SKU tissue-IHC caption: fixative not stated). The evidence does not establish whether frozen sections or IF are easier than paraffin IHC; leukocyte-associated peroxidase may cause background in appendix sections (standard IHC practice).
HPA tissue IHC evidence for CKS2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Caution, targets protein from more than one gene. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: CKS2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced CKS2 IHC Tips
Troubleshoot CKS2 staining in paraffin sections by checking retrieval, controls, compartment assignment, and scoring before interpreting biological differences.
What retrieval should I try when CKS2 staining is weak?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval setting). Allow sections to cool in the retrieval buffer, then compare a documented positive tissue with an adjacent section processed without primary antibody (standard IHC practice; HPA: high staining in bone marrow hematopoietic cells). Keep section thickness, detection chemistry, and development time constant while adjusting retrieval, so changes in signal remain interpretable (standard IHC practice). If staining remains weak, test a more alkaline retrieval buffer on matched sections as a fallback, watching for tissue damage and increased background (standard IHC practice).
Could fixation explain inconsistent CKS2 staining between paraffin blocks?
The supplied tissue caption identifies paraffin embedded human brain but does not report its fixative, so CKS2 specific fixation sensitivity is unknown (A08202-1 tissue caption). Record each block’s fixative, fixation duration, processing history, and storage conditions before comparing staining intensity (standard IHC practice). Run the blocks together with the same citrate pH 6.0 retrieval at 95–98 °C for 20 min and identical detection conditions (page retrieval setting; standard IHC practice). If differences track block preparation, report that limitation and assess preservation with routine morphology and a suitable control stain before attributing them to CKS2 biology (standard IHC practice).
How should I judge nuclear versus cytoplasmic CKS2 staining?
Score nuclear and cytoplasmic staining separately: tissue IHC is described as nuclear in most tissues, while subcellular IF mainly shows cytosol and mitochondria, with additional nucleoplasm and vesicles (HPA tissue IHC; HPA subcellular). UniProt supplies no subcellular annotation for CKS2, so neither pattern alone establishes the true compartment in a particular section (UniProt P33552 subcellular record). Compare signal with intact cell morphology and an adjacent section lacking primary antibody, and record the cell type alongside the compartment (standard IHC practice). Treat compartment conclusions cautiously because the HPA antibody evidence carries a warning about recognition of proteins from multiple genes (HPA reliability; HPA subcellular).
Could an isoform or modified epitope explain uneven staining?
The supplied CKS2 record lists 0 isoforms, a 1–79 chain, no signal peptide or transmembrane segment, and one example modified residue, acetyllysine at position 4 (UniProt P33552 record). Those annotations do not identify the catalog antibody’s epitope or demonstrate that acetylation changes its binding (UniProt P33552 record; A08202-1 tissue caption). Compare matched sections across retrieval conditions and, if available, use an independent antibody recognizing a documented different epitope to investigate discordant staining (standard IHC practice). The peptide blocked brain image supports competition under that image’s conditions, but peptide competition alone cannot establish which endogenous protein produced every signal (A08202-1 tissue caption; standard IHC interpretation).
How can IF help investigate an unexpected CKS2 IHC pattern?
Use IF as a separate validation experiment and multiplex CKS2 with a marker for the cell type being assessed, such as a hematopoietic cell marker when examining bone marrow (HPA: high in bone marrow hematopoietic cells; standard IF practice). Choose fluorophores after checking tissue autofluorescence and reserve a well separated channel for the weaker signal (standard IF practice). The record reports no transmembrane segment, while HPA IF describes cytosol, mitochondria, nucleoplasm, and vesicles; permeabilise adequately for intracellular epitopes without assuming a known antibody epitope (UniProt P33552 topology; HPA subcellular; standard IF practice). Include single colour and no primary controls before comparing IF compartments with chromogenic IHC (standard IF practice).
What should I change if CKS2 staining is diffuse or widespread?
Inspect a no primary section first to distinguish antibody associated staining from detection background (standard IHC practice). For chromogenic detection, check endogenous peroxidase blocking, reduce excessive DAB development, and optimise protein blocking and washes while keeping the positive control in the run (standard IHC practice). Titrate the IHC validated antibody empirically because the supplied evidence gives no working dilution; compare each dilution on the same tissue and at the same development time (A08202-1 tissue caption; standard IHC practice). Widespread nuclear signal is plausible from the HPA tissue profile, but that profile carries a cross gene antibody caution and cannot by itself certify diffuse staining as specific (HPA tissue IHC; HPA reliability).
How should I score CKS2 across samples with different cellularity? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because HPA tissue IHC describes predominantly nuclear staining while HPA IF reports several additional compartments (HPA tissue IHC; HPA subcellular). For nuclear chromogenic staining, report the percentage of eligible cells at each intensity and, when useful, an H-score from 0–300 (standard IHC scoring practice). Alternatively, report positive cell density per mm² of viable, evaluated tissue, excluding folds and necrosis using the same rules for every section (standard IHC scoring practice). Normalise positive counts to the number of eligible cells, or density to evaluated area, and document thresholds and control performance before comparing groups (standard IHC scoring practice).
When is apparent CKS2 positivity likely to be artefactual?
Interpret signal in intact, identifiable cells and compare its compartment with the nuclear tissue IHC profile and the cytosol, mitochondrial, nucleoplasm, and vesicle IF observations (HPA tissue IHC; HPA subcellular). Flag staining confined to section edges, folds, or necrotic areas, or persisting without primary antibody, as suspect (standard IHC interpretation). Check endogenous peroxidase activity when DAB appears in a no primary control, and compare the same cell type across sections before calling an unexpected population positive (standard IHC practice). The peptide blocked paraffin brain image provides a useful comparison, but HPA’s cross gene warning means concordant staining alone does not prove CKS2 specificity (A08202-1 tissue caption; HPA reliability).
Boster reagents
Best CKS2 / Cyclin-dependent kinases regulatory subunit 2 IHC Antibodies
The catalog lists a human- and mouse-reactive anti-CKS2 antibody for IHC and IF/ICC (catalog: A08202-1 applications and reactivity); its supplied image shows paraffin-section IHC, with no IF image supplied (catalog: image captions).
Real IHC data
Anti-CKS2 Antibody
Cat # A08202-1
A08202-1 will render with an IHC image of paraffin-embedded human brain tissue and a peptide-blocked comparison (catalog: A08202-1 IHC image caption). IHC, IF, and ICC are listed applications, and Human and Mouse are listed reactivities; no IF image is supplied (catalog: A08202-1 applications, reactivity, and image captions).
Which to pick: For tissue IHC, choose A08202-1: its own image documents staining of paraffin-embedded human brain tissue; the fixative is unreported (catalog: A08202-1 IHC image caption). For IF/ICC, A08202-1 lists both applications but has no supplied IF image (catalog: A08202-1 applications and image captions). For human or mouse samples, A08202-1 is the only listed option and is rabbit polyclonal; the supplied tissue image documents human tissue only (catalog: A08202-1 reactivity, host, dilution_raw, and IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P33552 (CKS2_HUMAN, Cyclin-dependent kinases regulatory subunit 2).
- Human Protein Atlas. CKS2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. CKS2 subcellular location (ICC-IF): Mainly localized to the cytosol and mitochondria. In addition localized to the nucleoplasm and vesicles. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. CKS2 antibody validation summary (2 antibodies).
- Cyclin-Dependent Kinase Subunit 2 (CKS2) as a Prognostic Marker for Stages I-III Invasive Non-Mucinous Lung Adenocarcinoma and Its Role in Affecting Drug Sensitivity. Cells 2022 — PMC9406629.
- CKS2 Overexpression Correlates with Prognosis and Immune Cell Infiltration in Lung Adenocarcinoma: A Comprehensive Study based on Bioinformatics and Experiments. Journal of Cancer 2021 — PMC8558665.
- CKS2 (CDC28 protein kinase regulatory subunit 2) is a prognostic biomarker in lower grade glioma: a study based on bioinformatic analysis and immunohistochemistry. Bioengineered 2021 — PMC8806895.
- Mitochondrial Function of CKS2 Oncoprotein Links Oxidative Phosphorylation with Cell Division in Chemoradioresistant Cervical Cancer. Neoplasia (New York, N.Y.) 2019 — PMC6411633.
- PubMed PMID:2227411 — UniProt-cited evidence.
- PubMed PMID:15164053 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CKS2 (Cyclin-dependent kinases regulatory subunit 2). Expected localisation Nuclear staining in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A08202-1 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); 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 brain tissue; fixative not specified (datasheet A08202-1); 2. Fixation: Image fixative and duration unreported (datasheet A08202-1); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 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-CKS2, 1:100 - 1:300 (datasheet A08202-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Caveat: Tissue IHC may detect protein from more than one gene (HPA tissue IHC); Isoform / epitope: 0 annotated isoforms; one 1–79 chain (UniProt); Regulation: Tissue-enhanced RNA: bone marrow, testis (HPA tissue RNA). Evidence: UniProt P33552, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol and mitochondria. In addition…, Datasheet A08202-1, PMC9406629, PMC8558665. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CKS2/immunohistochemistry-ihc-guide.