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CDK2 / Cyclin-dependent kinase 2 · IHC design guide
Design Immunohistochemistry for CDK2
Plan chromogenic CDK2 IHC in paraffin sections using the nuclear staining observed in subsets of intestinal, squamous and germinal-center cells as a reference (HPA tissue IHC). Consider epitope coverage across the 2 annotated isoforms when selecting an antibody (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CDK2 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 subsets of cells (HPA tissue IHC) | |
| Staining pattern | Nuclear staining in intestinal, squamous and germinal-center cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M00166-4) | |
| 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 paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M00166-4) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | No tissue-specific expression pattern is annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage needs checking (UniProt) |
Section 1
Recommended CDK2 IHC & IF Protocols
The catalog antibody protocol (datasheet: M00166-4) is followed by four published CDK2 IHC protocols (PMC11831524; PMC2816830; PMC3441142; PMC2668819).
| Sample | Paraffin-embedded human laryngeal squamous cell carcinomas tissue; fixative not specified (datasheet M00166-4) |
| Fixation | Image fixative and duration unreported (datasheet M00166-4); 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 M00166-4); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M00166-4) |
| Primary antibody | Mouse monoclonal (clone 6D5B5) anti-CDK2, 2 μg/ml (datasheet M00166-4) |
| Primary incubation | Overnight at 4 °C (datasheet M00166-4) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M00166-4) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CDK2-positive staining in lymphoid tissue of appendix (HPA tissue IHC: Medium). HPA tissue profile: Nuclear expression in subsets of cells in the intestinal tract and squamous epithelia, cells in germinal centers and seminiferous ducts. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet: M00166-4); use each published retrieval condition with its corresponding protocol (PMC3441142; PMC2668819).
Section 2
What Is the Expected CDK2 Staining Pattern?
CDK2 staining in paraffin sections should be predominantly nuclear in subsets of intestinal glandular cells, squamous epithelial cells, germinal center cells and cells of seminiferous ducts (HPA tissue IHC). HPA rates the tissue pattern Enhanced, with medium consistency between staining and RNA expression (HPA tissue IHC). CDK2 can also occupy cytoplasm and centrosomes; it has no transmembrane segment (UniProt P24941 localization and topology).
| Subset-restricted nuclear staining in intestinal glands, squamous epithelium or germinal centers. | This matches the reported tissue pattern (HPA tissue IHC). Duodenal and rectal glandular cells, esophageal and oral squamous cells, and lymph-node germinal center cells each have Medium staining (HPA tissue IHC). Score the named cell population and its nuclear signal rather than treating the entire tissue section as uniformly positive (HPA tissue IHC; general IHC practice). |
| Strong, widespread membrane-only or extracellular staining with little nuclear signal. | This differs from the reported nuclear tissue pattern (HPA tissue IHC). CDK2 has no transmembrane segment (UniProt P24941 topology). Review morphology and controls before calling it CDK2: HPA also reports a plasma-membrane location in ICC-IF, but marks that location uncertain (HPA subcellular ICC-IF). |
| Prominent staining of an HPA-listed negative cell population, such as adipocytes. | Adipocytes in adipose tissue and breast are reported as Not detected (HPA tissue IHC). Check whether the signal follows tissue edges or appears in the negative control; cross-reactivity or detection background is possible (general IHC practice). A positive result elsewhere in the section does not validate staining in adipocytes (general IHC practice). |
| Diffuse color across nuclei, cytoplasm and surrounding tissue, obscuring cell boundaries. | The observed tissue pattern is nuclear in subsets, so an indiscriminate haze is difficult to interpret (HPA tissue IHC). Cytoplasmic CDK2 is biologically possible (UniProt P24941 localization); cytoplasmic color alone is therefore insufficient to label a section artefactual. Use background controls and preserved morphology to distinguish a cellular pattern from diffuse detection background (general IHC practice). |
| No nuclear staining in lymph-node germinal centers or esophageal squamous cells. | Both populations have Medium staining in the HPA tissue record (HPA tissue IHC). First check that the expected cells are present and interpretable, then assess assay performance with a known-positive section and detection controls (general IHC practice). Absence in one field cannot establish CDK2 absence throughout a heterogeneous tissue (HPA tissue IHC). |
💡Expected CDK2 appearanceCall a section positive when a recognizable subset of the expected cells shows nuclear staining, often Medium in the listed positive populations; uniform extracellular color or staining confined to HPA-listed negative cells warrants control review (HPA tissue IHC; general IHC practice).
| Cell population and intensity | HPA reports Medium staining in several named epithelial and lymphoid populations, Low staining in placental cytotrophoblasts, and Not detected in specified cells such as adipocytes (HPA tissue IHC). Compare like cell types, not whole organs: a negative call for one cell population does not describe every cell in that tissue (HPA tissue IHC). |
| Compartment and cell-cycle context | HPA's tissue profile is nuclear in subsets (HPA tissue IHC). UniProt also lists cytoplasm, Cajal bodies, endosomes and centrosomes, with centrosomal localization in late G2 (UniProt P24941 localization). A small extranuclear signal can merit review, but localization alone does not establish kinase activity or cell-cycle phase (UniProt P24941 localization and function; general IHC practice). |
| Antibody validation and scope | The HPA tissue record is Enhanced, while its stated antibody-staining/RNA consistency is medium (HPA tissue IHC). CAB013115 is listed as IHC Enhanced; HPA066915 is listed as ICC Enhanced (HPA antibody validation). Apply the IHC evidence to paraffin-section expectations without treating the ICC rating as IHC validation (HPA antibody validation). |
| Q: What should IF/ICC show? | A: Mainly nucleoplasmic CDK2; centrosome and basal-body locations are supported, calyx is approved, and plasma-membrane location is uncertain (HPA subcellular ICC-IF). Those annotations describe ICC-IF localization, not a paraffin-section staining protocol or a requirement that every IHC-positive cell show each additional compartment (HPA subcellular ICC-IF; HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control tissue has no nuclear signal. | The expected cells may be absent from the field, or the staining run may have failed (HPA tissue IHC; general IHC practice). | Locate the named positive population, then verify retrieval, antibody incubation and chromogenic detection against the catalog antibody's IHC-P instructions (HPA tissue IHC; general IHC practice). |
| Every structure develops color. | Diffuse detection background or excess staining can mask the subset-restricted nuclear pattern (HPA tissue IHC; general IHC practice). | Compare a negative-control slide and review blocking, washes and chromogen development before interpreting cell localization (general IHC practice). |
| Adipocytes stain as strongly as germinal-center cells. | That distribution conflicts with the reported Not detected adipocytes and Medium germinal-center cells (HPA tissue IHC). | Inspect morphology and negative controls; report the unexpected population separately and confirm before calling it specific (general IHC practice). |
| Signal appears only at tissue edges or in extracellular spaces. | Such a distribution differs from the reported nuclear pattern (HPA tissue IHC). | Check for section-edge artefact and compare the same areas with a negative control; avoid scoring extracellular color as positive cells (general IHC practice). |
| Weak staining is seen in colon glands. | Colon glandular cells are listed as Low, whereas duodenal and rectal glandular cells are Medium (HPA tissue IHC). | Score the named cell type and compare with a listed medium-staining population on a separately controlled section before calling the run failed (HPA tissue IHC; general IHC practice). |
| Cytoplasmic signal accompanies nuclear staining. | Cytoplasm is a UniProt-listed CDK2 location, although HPA describes the tissue IHC pattern as nuclear in subsets (UniProt P24941 localization; HPA tissue IHC). | Record nuclear and cytoplasmic staining separately; use negative controls to assess the cytoplasmic component rather than assuming it is specific or artefactual (general IHC practice). |
Experiment evidence
IHC & IF images showing CDK2 staining
M00166-4 · IHCIHC analysis of Cdk2 using anti-Cdk2 antibody (M00166-4). Cdk2 was detected in a paraffin-embedded section of human laryngeal squamous cell
M00166-4 · IHCIHC analysis of Cdk2 using anti-Cdk2 antibody (M00166-4). Cdk2 was detected in a paraffin-embedded section of human liver cancer tissue. Hea
M00166-3 · IHCIHC analysis of Cdk2 using anti-Cdk2 antibody (M00166-3). Cdk2 was detected in a paraffin-embedded section of human breast cancer tissue. He
M00166-3 · IHCIHC analysis of Cdk2 using anti-Cdk2 antibody (M00166-3). Cdk2 was detected in a paraffin-embedded section of human laryngeal squamous cell
PB9534 · IHCIHC analysis of Cdk2 using anti-Cdk2 antibody (PB9534). Cdk2 was detected in a paraffin-embedded section of human invasive urothelial carcin
PB9534 · IFIF analysis of Cdk2 using anti-Cdk2 antibody (PB9534) and anti-Beta Tubulin antibody (M01857-3). Cdk2 was detected in immunocytochemical sec
M00166 · IHCImmunohistochemical analysis of paraffin-embedded human beast carcinoma, using CDK2 Antibody.
M00166 · IHCImmunohistochemical analysis of paraffin-embedded Rat heart, using the Antibody at 1:200 dilution.
M00166 · IFImmunofluorescent analysis of Hela cells, using CDK2 Antibody .
M00166-6 · IHCHuman tonsil was stained with anti-CDK2 rabbit antibody
MP00166 · IHCImmunohistochemical analysis of paraffin-embedded human bladder cancer, using Phospho-CDK2 (Y15) Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000123374-CDK2/tissue/cerebral+cortex#imid_3669448
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000123374-CDK2/tissue/colon#imid_3669366
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000123374-CDK2/tissue/liver#imid_3669367
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000123374-CDK2/tissue/kidney#imid_3669360
HPA · ICC-IF SK-MEL-30Human Protein Atlas immunofluorescence, SK-MEL-30, antibody HPA066915; green = CDK2, blue = nucleus, red = microtubules.
HPA · ICC-IF SK-MEL-30Human Protein Atlas immunofluorescence, SK-MEL-30, antibody HPA066915; green = CDK2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CDK2 IHC & IF
🧪Run lymph node first and score germinal center cells as the expected positive population (Medium; HPA: lymph node, germinal center cells). Use adipose tissue adipocytes as the negative comparator (HPA: adipose tissue, adipocytes Not detected); on the positive slide, cells without specific nuclear or cytoplasmic staining can serve as internal negatives if present, but their identity and negativity must be verified rather than assumed (UniProt P24941: nuclear and cytoplasmic localization; standard IHC practice).
Positive control tissue: Appendix (Lymphoid tissue, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CDK2 in SK-MEL-30, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), Sperm, HeLa BAC 5399, HeLa , with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a mouse IgG isotype control matched to the primary antibody’s class and, if monoclonal, subclass (selected-SKU caption: mouse anti-CDK2 primary and goat anti-mouse secondary; standard IHC practice). Use a matched CDK2 knockout sample as a biological negative if available, and quench endogenous peroxidase in the lymph node section before HRP/DAB detection (standard IHC practice).
⚠️Feasibility: A CDK2-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state its fixative (selected-SKU caption: fixative not stated). The demonstrated paraffin-section procedure uses heat retrieval in EDTA at pH 8.0, but the evidence does not establish that CDK2 staining requires this retrieval condition (selected-SKU caption: EDTA heat retrieval). HPA documents ICC-IF localization, but the supplied evidence does not establish that IF or frozen sections are easier than paraffin IHC; assess leukocyte-associated endogenous peroxidase background when scoring lymph node HRP/DAB staining (HPA: ICC-IF subcellular data; standard IHC practice).
HPA tissue IHC evidence for CDK2
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 | Lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Basal cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CDK2 IHC Tips
Troubleshoot CDK2 staining in paraffin section IHC by checking retrieval, cell type, compartment, and controls before interpreting signal.
What retrieval should I use when CDK2 staining is weak in paraffin sections?
Start with heat mediated antigen retrieval in EDTA at pH 8.0 (datasheet M00166-4). The selected antibody stained a paraffin section after this retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet M00166-4). If signal is weak, compare retrieval heating and cooling across adjacent sections while holding antibody concentration and DAB development constant (standard IHC practice). Judge improvement in viable cells and expected compartments against a no primary control; stronger edge staining alone does not establish improved CDK2 detection (HPA: nuclear expression in cell subsets; standard IHC practice).
Can the available evidence tell me whether fixation has masked CDK2?
Target specific CDK2 sensitivity to fixative type or fixation duration is unknown from the supplied evidence (datasheet M00166-4: fixative not stated). The selected image shows staining in a paraffin embedded human laryngeal squamous cell carcinoma section, but its caption does not identify the fixative (datasheet M00166-4). Record the fixative and fixation duration for each block, then compare sections processed with the same EDTA pH 8.0 retrieval and detection settings (datasheet M00166-4; standard IHC practice). If staining varies, check tissue preservation and processing history before attributing the difference to CDK2 biology or a specific fixation effect (standard IHC practice).
Which CDK2 staining compartments are plausible, and when should I investigate a mismatch?
Assess nuclear staining first, particularly in cell subsets, because tissue IHC reports nuclear CDK2 in squamous epithelia, intestinal tract, germinal centers and seminiferous ducts (HPA: tissue IHC). Cytoplasmic or centrosomal signal can also be plausible: CDK2 is annotated in the cytoplasm and centrosome, with centrosomal localization in late G2 (UniProt P24941: subcellular location). Score nucleus and cytoplasm separately rather than treating every stained pixel as equivalent (standard IHC practice). Diffuse extracellular or luminal DAB deserves investigation because CDK2 has no transmembrane segment or secretory signal, while tissue folds and precipitate can mimic staining (UniProt P24941: topology and processing; standard IHC practice).
Could isoforms or modified residues explain discordant CDK2 IHC staining?
CDK2 has 2 annotated isoforms, but the supplied antibody caption does not identify its epitope or establish isoform specific recognition (UniProt P24941: isoforms; datasheet M00166-4). The canonical chain spans residues 1–298, and its kinase domain spans residues 4–286 (UniProt P24941: processing and domains). Annotated modifications include acetylation and phosphorylation near the amino terminus, so epitope accessibility is a hypothesis to test, not an established cause of staining differences (UniProt P24941: modified residues; standard IHC practice). Request epitope mapping or compare an independently validated antibody before assigning a compartment or intensity difference to an isoform or modification (standard IHC practice).
How should I troubleshoot CDK2 IF alongside this chromogenic IHC guide?
Treat IF/ICC as a separate validation exercise: the selected antibody evidence documents paraffin section chromogenic IHC, with no IF/ICC result supplied for that antibody (datasheet M00166-4). For multiplexing, pair CDK2 with a marker identifying the expected cell population and examine nuclear signal within that population; squamous epithelial and germinal center subsets are tissue IHC examples (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). Because CDK2 is intracellular and has no transmembrane segment, verify that permeabilisation exposes nuclear and cytoplasmic epitopes without disrupting morphology (UniProt P24941: subcellular location and topology; standard IF practice).
How can I distinguish excess DAB background from weak but genuine CDK2 staining?
Run a no primary control and inspect blank tissue regions for nonspecific DAB, precipitate, and endogenous peroxidase activity (standard IHC practice). The selected IHC caption used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase conjugated secondary for 30 minutes at 37°C (datasheet M00166-4). If background rises, check wash consistency, antibody concentration and DAB development time one variable at a time; use a peroxidase block as a general chromogenic IHC control (standard IHC practice). Preserve cell specific nuclear staining while optimizing, since reported tissue expression occurs in subsets rather than uniformly across every cell (HPA: tissue IHC).
What should I score when comparing CDK2 across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define viable tissue regions and cell classes before scoring, because reported CDK2 staining is nuclear in subsets of specific cell populations (HPA: tissue IHC; standard IHC practice). Report the percentage of positive nuclei and, if intensity is reproducible, an H-score from 0–300 using intensity grades 0–3 (standard IHC practice). Normalize positive cell counts to the number of eligible cells in each annotated compartment, or report positive cell density per mm² of viable tissue (standard IHC practice). Keep retrieval, DAB development, imaging and scoring thresholds consistent; record cytoplasmic or centrosomal staining separately from the nuclear score (datasheet M00166-4: EDTA pH 8.0; UniProt P24941: subcellular location; standard IHC practice).
When does apparent CDK2 positivity reflect artefact rather than meaningful tissue staining?
Give greatest weight to staining in intact cells and plausible compartments, including nuclear subsets reported in squamous epithelia and germinal centers (HPA: tissue IHC). A centrosomal focus can be plausible in late G2, but CDK2 also has cytoplasmic annotations, so compartment alone cannot establish activity or cell cycle phase (UniProt P24941: subcellular location and function). Suspect artefact when DAB accumulates at section edges, tissue folds or necrotic regions, or appears similarly in a no primary control (standard IHC practice). Investigate diffuse extracellular deposits or persistent signal after omitting primary antibody, including endogenous peroxidase as a possible source (UniProt P24941: no secretory signal; standard IHC practice).
Boster reagents
Best CDK2 / Cyclin-dependent kinase 2 IHC Antibodies
Anti-CDK2 antibodies have IHC images from human and rat tissue (catalog IHC captions) and IF/ICC images from human cells (M00166 and PB9534 IF captions).
Real IHC data
Anti-Cdk2 Antibody ® (monoclonal, 6D5B5)
Cat # M00166-4
Real IHC data
Anti-Cdk2 Antibody ® (monoclonal, 5B12D1)
Cat # M00166-3
Real IHC data
Anti-Cdk2 Antibody ®
Cat # PB9534
Real IHC data
Anti-CDK2 Rabbit Monoclonal Antibody
Cat # M00166
Real IHC data
Anti-CDK2 Rabbit Monoclonal Antibody
Cat # M00166-6
Real IHC data
Anti-Phospho-CDK2 (Y15) Rabbit Monoclonal Antibody
Cat # MP00166
M00166-4 was imaged in human laryngeal carcinoma, M00166-3 in human breast cancer, and PB9534 in human bladder carcinoma (respective IHC captions). M00166 was imaged in human breast carcinoma and rat heart, M00166-6 in human tonsil, and phospho-CDK2 (Y15) antibody MP00166 in human bladder cancer (respective IHC captions; MP00166 catalog title).
Which to pick: For paraffin-section IHC, choose mouse monoclonal M00166-4: its own caption documents EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody; the fixative is unreported (M00166-4 catalog entry and IHC caption). For IF/ICC, consider rabbit monoclonal M00166 or rabbit PB9534; both list IF/ICC and have human-cell IF images (M00166 and PB9534 catalog entries and IF captions). For work spanning human and rat tissue, M00166 has IHC images in both, while mouse reactivity is listed in the catalog without a mouse IHC image in the supplied captions (M00166 catalog reactivity and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P24941 (CDK2_HUMAN, Cyclin-dependent kinase 2).
- Human Protein Atlas. CDK2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CDK2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the plasma membrane, centrosome, basal body and calyx..
- Human Protein Atlas. CDK2 antibody validation summary (2 antibodies).
- Targeting CDK2 Confers Vulnerability to Lenvatinib Via Driving Senescence in Anaplastic Thyroid Cancer. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2025 — PMC11831524.
- Expression of cell cycle regulators during smooth muscle cell proliferation after balloon catheter injury of rat artery. Journal of Korean medical science 2004 — PMC2816830.
- Involvement of Akt, Ras and cell cycle regulators in the potential development of endometrial hyperplasia in women with polycystic ovarian syndrome. Gynecologic oncology 2009 — PMC3441142.
- Genetic substitution of Cdk1 by Cdk2 leads to embryonic lethality and loss of meiotic function of Cdk2. Development (Cambridge, England) 2008 — PMC2668819.
- PubMed PMID:1714386 — UniProt-cited evidence.
- PubMed PMID:1653904 — UniProt-cited evidence.
- PubMed PMID:1717994 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CDK2 (Cyclin-dependent kinase 2). Expected localisation Nuclear staining in subsets of cells (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M00166-4). Validated IHC antibody: M00166-4 at 2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Lymphoid tissue); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human laryngeal squamous cell carcinomas tissue; fixative not specified (datasheet M00166-4); 2. Fixation: Image fixative and duration unreported (datasheet M00166-4); 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 M00166-4); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M00166-4); 8. Primary antibody: Mouse monoclonal (clone 6D5B5) anti-CDK2, 2 μg/ml (datasheet M00166-4); 9. Primary incubation: Overnight at 4 °C (datasheet M00166-4). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M00166-4); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage needs checking (UniProt); Regulation: No tissue-specific expression pattern is annotated (UniProt). Evidence: UniProt P24941, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet M00166-4, PMC11831524, PMC2816830. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CDK2/immunohistochemistry-ihc-guide.