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CDKN1C / Cyclin-dependent kinase inhibitor 1C · IHC design guide
Design Immunohistochemistry for CDKN1C
Plan CDKN1C chromogenic IHC in paraffin sections using nuclear staining as the expected pattern (HPA tissue IHC). The guide identifies high-staining reference populations, including kidney glomerular cells and placental decidual cells (HPA tissue IHC), and helps assess cell-specific results.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CDKN1C 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 several tissues (HPA tissue IHC) | |
| Staining pattern | Nuclear staining in selected cell populations (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01244-2) | |
| Positive control | Adrenal gland+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 conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining is cell-specific; lung alveolar cells were not detected (HPA tissue IHC) | |
| Regulation | Expression varies by tissue and cell type (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms, Long and Short; check epitope coverage (UniProt) |
Section 1
Recommended CDKN1C IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A01244-2) is followed by published CDKN1C staining protocols for human breast cancer and mouse fetal sections (PMC2323395; PMC4786089).
| Sample | Paraffin-embedded human esophageal squamous carcinoma tissue; fixative not specified (datasheet A01244-2) |
| Fixation | Image fixative and duration unreported (datasheet A01244-2); 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 A01244-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01244-2) |
| Primary antibody | Rabbit anti-CDKN1C, 2 μg/ml (datasheet A01244-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01244-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01244-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CDKN1C-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet: A01244-2); the published protocols used citrate retrieval with their respective antibodies (PMC2323395; PMC4786089).
Section 2
What Is the Expected CDKN1C Staining Pattern?
CDKN1C should appear mainly in nuclei on paraffin-section IHC: UniProt places it in the nucleus and reports no transmembrane segment (UniProt P49918 topology). Expect staining in adrenal glandular cells, glomerular cells, pancreatic endocrine cells and placental decidual cells (HPA: High). HPA describes nuclear expression across several tissues; its IHC reliability is Enhanced, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC).
| Distinct nuclear staining in the expected cell population, strongest among the supplied examples in adrenal glandular, glomerular, pancreatic endocrine or decidual cells (HPA: High). | This fits the reported IHC pattern (HPA: nuclear expression; High in these cell populations). Assess the named cells within each tissue: a positive section does not imply that every cell in it should stain. |
| Predominantly cytoplasmic staining, or staining outside cells, with little convincing nuclear signal. | For paraffin-section IHC, this conflicts with the reported nuclear pattern (UniProt P49918: nucleus; HPA: tissue IHC). Review staining specificity and detection artefacts (general IHC practice). Cytosolic ICC-IF signal is reported separately (HPA: subcellular ICC-IF). |
| Strong staining in a cell population reported as not detected, such as adipocytes or lung alveolar cells (HPA: Not detected). | Check whether the stained cells match the named HPA population, then investigate cross-reactivity or endogenous detection activity (general IHC practice). An HPA “Not detected” result is a reference observation, not proof that every specimen must be negative. |
| Broad haze across tissue compartments, including areas without a discernible nuclear pattern. | This does not resemble the reported nuclear IHC pattern (HPA: tissue IHC). Uneven blocking, insufficient washing or detection-system background can obscure a specific signal (general IHC practice); compare the slide with its detection controls. |
| No nuclear signal in adrenal glandular cells, glomerular cells, pancreatic endocrine cells or decidual cells (HPA: High). | A negative result in one of these reference populations warrants checking tissue identity, cell preservation and the IHC workflow (general IHC practice). HPA levels describe observed staining; they do not guarantee the same intensity in every specimen. |
💡Expected CDKN1C appearanceCall positive when staining is chiefly nuclear in the specified glandular, glomerular, endocrine or decidual cells, with conspicuous signal in HPA High examples; diffuse haze or isolated staining in an HPA Not detected population needs investigation (HPA: tissue IHC).
| Cell population and tissue context | HPA reports High staining in four specified populations, Medium in several others, and Not detected in others (HPA: tissue IHC). Score the relevant cell population rather than assigning one intensity to an entire section. |
| Strength of the tissue-IHC evidence | The tissue profile is marked Enhanced, while antibody staining and RNA show medium consistency (HPA: tissue IHC). These labels support a reference pattern but do not turn RNA abundance into a per-cell staining prediction. |
| Antibody validation scope | HPA002924 is listed as IHC Enhanced and ICC Approved (HPA: antibody validation). Those are application-specific validation labels; they do not establish the performance of another antibody or a particular specimen. |
| IF/ICC Q: Where might signal appear? | A: Mainly in nucleoplasm and nuclear bodies, with additional cytosolic localization (HPA: subcellular ICC-IF). This cell-based observation should not be used to require diffuse cytoplasmic staining in paraffin-section IHC. |
| Isoforms and epitope uncertainty | UniProt lists Long and Short isoforms (UniProt P49918: isoforms). Without an epitope position in the supplied record, whether an antibody detects both cannot be determined; avoid attributing a weak result to one isoform. |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected positive population has no nuclear stain. | The observed result differs from HPA High staining in the named population (HPA: tissue IHC); tissue selection, slide condition or a workflow step may account for it (general IHC practice). | Confirm the correct cells are present; inspect a suitable positive reference and review retrieval, primary-antibody dilution and detection against the antibody's validated IHC instructions (general IHC practice). |
| Signal is chiefly cytoplasmic or extracellular on IHC. | That distribution conflicts with nuclear tissue IHC (HPA: tissue IHC; UniProt P49918: nucleus). ICC-IF reports some cytosolic localization, which does not by itself validate this IHC pattern (HPA: subcellular ICC-IF). | Compare nuclear detail and control slides; review antibody specificity, washing and detection background before interpreting the signal as CDKN1C (general IHC practice). |
| Adipocytes or lung alveolar cells stain strongly. | Those particular populations were Not detected in HPA tissue IHC (HPA: adipocytes; lung alveolar cells). Misidentified cells, cross-reactivity or endogenous detection activity are possible explanations (general IHC practice). | Verify cell identity and compare positive and negative reference areas; inspect detection controls for background activity (general IHC practice). |
| The whole section has diffuse chromogen or uneven haze. | Nonspecific background can hide the reported nuclear pattern (HPA: tissue IHC); blocking, washing and detection conditions are general sources to examine (general IHC practice). | Check a no-primary control, blocking and wash steps, then read only cell-associated nuclear signal above the control background (general IHC practice). |
| A reference-positive area stains more weakly than expected. | HPA levels are observational categories, not calibrated intensity targets (HPA: tissue IHC). Variation in the sampled cell population or assay conditions can affect comparisons (general IHC practice). | Compare the same named cell population across sections, confirm the IHC conditions used, and record intensity with the control result rather than imposing an HPA score on the specimen (general IHC practice). |
| A different primary antibody gives a different pattern. | The supplied validation entry applies to HPA002924 (HPA: antibody validation); the record does not establish another antibody's epitope or isoform coverage (UniProt P49918: Long and Short isoforms). | Check that antibody's IHC validation and epitope information, then compare staining in the HPA High reference populations and appropriate controls (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing CDKN1C staining
A01244-2 · IHCIHC analysis of p57 Kip2/CDKN1C using anti-p57 Kip2/CDKN1C antibody (A01244-2). p57 Kip2/CDKN1C was detected in a paraffin-embedded section
A01244-2 · IHCIHC analysis of p57 Kip2/CDKN1C using anti-p57 Kip2/CDKN1C antibody (A01244-2). p57 Kip2/CDKN1C was detected in a paraffin-embedded section
M01244-2 · IHCImmunohistochemical analysis of paraffin-embedded Human thyroid cancer, using the Antibody at 1:50 dilution.
M01244-2 · IHCImmunohistochemical analysis of paraffin-embedded Human ovarian cancer, using the Antibody at 1:50 dilution.
M01244-2 · IFImmunofluorescent analysis of HeLa cells treated with dexamethasone, using p57 Kip2 Antibody .
M01244-3 · IHCHuman kidney tissue was stained with anti-p57kip2 rabbit Antibody
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000129757-CDKN1C/tissue/cerebral+cortex#imid_19229335
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000129757-CDKN1C/tissue/colon#imid_19229269
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000129757-CDKN1C/tissue/liver#imid_19229262
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000129757-CDKN1C/tissue/kidney#imid_19229263
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA002924; green = CDKN1C, blue = nucleus, red = microtubules.
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA002924; green = CDKN1C, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CDKN1C IHC & IF
🧪Run placenta first: decidual-cell nuclei should stain (HPA: High in decidual cells; UniProt P49918: nucleus). Use adipose tissue as a negative, with adipocytes at background (HPA: Not detected in adipocytes); on the positive slide, assess other cells for background staining without assuming they are CDKN1C-negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CDKN1C in HeLa, SiHa, U2OS, with annotated localisation: Nucleoplasm (approved), Nuclear bodies (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a concentration-matched rabbit IgG isotype control for the captioned rabbit antibody, and a validated CDKN1C knockout or peptide-block control (caption: rabbit antibody; standard IHC practice). Check placental sections for endogenous peroxidase and biotin background before interpreting DAB signal, especially with the captioned biotin–streptavidin detection (caption: biotinylated secondary, SABC and DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A01244-2 paraffin-section caption does not state the fixative (caption: fixative unreported). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required; no matched evidence establishes that frozen sections or IF are easier (caption: EDTA retrieval; supplied evidence). ICC-IF images exist for HeLa, SiHa and U2OS, while placental endogenous peroxidase or biotin can complicate chromogenic interpretation (HPA: ICC-IF image cell lines; standard IHC practice).
HPA tissue IHC evidence for CDKN1C
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | High | Protein (IHC) | HPA → |
| Pancreas | Pancreatic endocrine cells | High | Protein (IHC) | HPA → |
| Placenta | Decidual cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Colon | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CDKN1C IHC Tips
Troubleshoot CDKN1C chromogenic IHC in paraffin sections by checking retrieval, nuclear localisation, cell type and assay controls before interpreting staining.
What retrieval should I try first when CDKN1C nuclear staining is weak?
Start with heat-mediated retrieval in EDTA, pH 8.0, the condition used for the catalog antibody in a paraffin section (datasheet A01244-2). Keep retrieval time and heating conditions consistent across slides while comparing signal in a control with expected nuclear staining, such as placental decidual cells (HPA: High in decidual cells). The caption reports 2 μg/ml primary antibody overnight at 4°C, so check those variables before attributing weak staining to retrieval (datasheet A01244-2). If staining remains weak, test a second retrieval condition on matched sections as a troubleshooting experiment, and record both nuclear signal and tissue damage (standard IHC practice).
Could fixation explain weak or uneven CDKN1C staining in my paraffin sections?
Target-specific fixation sensitivity for CDKN1C is unknown from the supplied evidence; the catalog image identifies a paraffin section but does not state its fixative (datasheet A01244-2). Record the fixative, fixation duration and processing history for each specimen before comparing staining intensity across cases (standard IHC practice). Examine whether weak nuclear signal tracks with poorly preserved morphology or varies across a section, and repeat staining on a better-preserved section when available (standard IHC practice). Keep retrieval at EDTA, pH 8.0 during that comparison so a change in retrieval does not obscure a processing effect (datasheet A01244-2).
How should I evaluate CDKN1C staining outside the nucleus?
Prioritise nuclear staining when reading chromogenic CDKN1C IHC: UniProt assigns CDKN1C to the nucleus, and HPA reports nuclear expression in several tissues (UniProt P49918; HPA: tissue IHC). HPA cell imaging places it mainly in the nucleoplasm and nuclear bodies, with an additional cytosolic location, so cytoplasmic colour alone is insufficient for a tissue-level positive call (HPA: subcellular). Compare the stained compartment with the hematoxylin counterstain at high magnification, and check whether nuclei in an expected positive cell population stain consistently (standard IHC practice). CDKN1C has no annotated transmembrane segment, so a crisp membrane-only pattern needs independent verification (UniProt P49918 topology).
Could CDKN1C isoforms or epitope masking explain discordant staining?
CDKN1C has 2 reported isoforms, Long and Short, but the supplied catalog caption does not identify the antibody epitope or establish which isoforms it detects (UniProt P49918; datasheet A01244-2). Ask for epitope information before treating an isoform difference as the explanation for discordant tissue staining (standard IHC practice). UniProt lists methylation at residue 107 and phosphorylation at residue 268; their effect on this antibody’s binding has not been established here (UniProt P49918). Compare matched sections under the stated EDTA, pH 8.0 retrieval condition, then assess antibody specificity with an independent method if the disagreement persists (datasheet A01244-2; standard IHC practice).
How can IF help resolve which cells carry a CDKN1C signal?
Use IF as a separate follow-up assay to map CDKN1C alongside a marker for the expected cell type, then compare its cellular assignment with the chromogenic IHC result (standard IF practice). For example, HPA reports high CDKN1C staining in pancreatic endocrine cells, so a validated endocrine-cell marker can help assess that population (HPA: High in pancreatic endocrine cells). Choose fluorophores after checking tissue autofluorescence and include single-label controls to distinguish bleed-through from overlap (standard IF practice). Because CDKN1C is mainly nuclear and has no transmembrane segment, plan permeabilisation for an intracellular epitope, while establishing fixation and antibody conditions specifically for IF (HPA: subcellular; UniProt P49918 topology; standard IF practice).
What should I check when DAB appears broadly across the section?
Separate diffuse chromogen deposition from cell-specific nuclear staining by examining a no-primary control alongside the test slide (standard IHC practice). The catalog image used 10% goat serum blocking, a biotinylated secondary for 30 minutes at 37°C, a streptavidin–biotin detection complex and DAB (datasheet A01244-2). Check the peroxidase block, washing and detection reagents as general chromogenic workflow steps if the no-primary control also develops colour (standard IHC practice). If background persists only with primary antibody, titrate around the reported 2 μg/ml condition and judge whether nuclear contrast improves in an expected positive control (datasheet A01244-2; standard IHC practice).
How should I score CDKN1C IHC across heterogeneous tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, then report the percentage of positive nuclei and nuclear intensity rather than whole-section DAB coverage (standard IHC practice; UniProt P49918 subcellular). An H-score combines percentages at intensity grades 0–3 into a 0–300 score; apply the same thresholds to every case (standard IHC practice). Alternatively, count positive nuclei per mm² and normalise to all assessable nuclei of the same cell type in the measured region (standard IHC practice). Record excluded necrotic and damaged areas, and use an appropriate positive reference because CDKN1C expression varies by tissue and cell population (standard IHC practice; HPA: tissue IHC).
How do I distinguish a true CDKN1C positive from staining artefact?
A credible positive shows reproducible nuclear staining in morphologically intact cells of an expected population, such as placental decidual cells or pancreatic endocrine cells (HPA: High in both populations; UniProt P49918 subcellular). Treat membrane-only colour cautiously because CDKN1C has no annotated transmembrane segment, while recognising that HPA also reports an additional cytosolic location (UniProt P49918 topology; HPA: subcellular). Check whether staining concentrates at section edges or in necrotic areas, and compare a no-primary control for endogenous enzyme or detection background (standard IHC practice). Interpret negative cells in context: HPA reports CDKN1C as undetected in some sampled cell populations, including adipocytes (HPA: Not detected in adipocytes).
Boster reagents
Best CDKN1C / Cyclin-dependent kinase inhibitor 1C IHC Antibodies
Anti-CDKN1C antibodies have IHC images from human and rat tissues (catalog IHC captions) and an IF image from HeLa cells (M01244-2 IF caption); mouse reactivity is listed (catalog reactivity).
Real IHC data
Anti-p57 Kip2/CDKN1C Antibody ®
Cat # A01244-2
Real IHC data
Anti-p57 Kip2 Rabbit Monoclonal Antibody
Cat # M01244-2
Real IHC data
Anti-p57kip2 Rabbit Monoclonal Antibody
Cat # M01244-3
A01244-2 has paraffin-section IHC images from human esophageal, cervical and lung cancers and rat kidney (A01244-2 IHC captions); M01244-2 has paraffin-section IHC images from human thyroid, ovarian and tongue cancers and rat kidney, plus an IF image from dexamethasone-treated HeLa cells (M01244-2 IHC and IF captions). M01244-3 has a human kidney IHC image, with section processing unreported (M01244-3 IHC caption).
Which to pick: For tissue IHC, choose A01244-2 when a documented starting procedure helps: its paraffin-section captions specify EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody, but do not report the fixative (A01244-2 IHC captions). For IF/ICC, choose monoclonal M01244-2, which lists both applications and has a HeLa IF image; its IHC captions document paraffin sections without reporting the fixative (M01244-2 catalog applications and IF/IHC captions). For cross-species IHC, A01244-2 and M01244-2 list human, mouse and rat reactivity, while M01244-3 lists human reactivity and has a human kidney IHC image (catalog applications/reactivity; M01244-3 IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P49918 (CDN1C_HUMAN, Cyclin-dependent kinase inhibitor 1C).
- Human Protein Atlas. CDKN1C tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CDKN1C subcellular location (ICC-IF): Mainly localized to the nucleoplasm and nuclear bodies. In addition localized to the cytosol..
- Human Protein Atlas. CDKN1C antibody validation summary (1 antibodies).
- CDKN1C/p57kip2 is a candidate tumor suppressor gene in human breast cancer. BMC cancer 2008 — PMC2323395.
- Cdkn1c Boosts the Development of Brown Adipose Tissue in a Murine Model of Silver Russell Syndrome. PLoS genetics 2016 — PMC4786089.
- Relationship of DNA methylation to mutational changes and transcriptional organization in fusion-positive and fusion-negative rhabdomyosarcoma. International journal of cancer 2019 — PMC7415348.
- Mutations in the PCNA-binding domain of CDKN1C cause IMAGe syndrome. Nature genetics 2012 — PMC3386373.
- PubMed PMID:7729684 — UniProt-cited evidence.
- PubMed PMID:8640800 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CDKN1C (Cyclin-dependent kinase inhibitor 1C). Expected localisation Nuclear staining in several tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01244-2). Validated IHC antibody: A01244-2 at 2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human esophageal squamous carcinoma tissue; fixative not specified (datasheet A01244-2); 2. Fixation: Image fixative and duration unreported (datasheet A01244-2); 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 A01244-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01244-2); 8. Primary antibody: Rabbit anti-CDKN1C, 2 μg/ml (datasheet A01244-2); 9. Primary incubation: Overnight at 4 °C (datasheet A01244-2). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining is cell-specific; lung alveolar cells were not detected (HPA tissue IHC); Isoform / epitope: 2 isoforms, Long and Short; check epitope coverage (UniProt); Regulation: Expression varies by tissue and cell type (HPA tissue IHC). Evidence: UniProt P49918, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm and nuclear bodies. In addition…, Datasheet A01244-2, PMC2323395, PMC4786089. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CDKN1C/immunohistochemistry-ihc-guide.