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CUEDC2 / CUE domain-containing protein 2 · IHC design guide
Design Immunohistochemistry for CUEDC2
Plan CUEDC2 paraffin IHC with the IHC-validated antibody at 0.5–1 µg/mL (datasheet: PA1962). Assess cytoplasmic staining in lung macrophages or glomerular cells (HPA tissue IHC), and interpret results cautiously because tissue IHC reliability is uncertain (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CUEDC2 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic tissue staining (HPA tissue IHC); nuclear location annotated (UniProt) | |
| Staining pattern | Cytoplasmic staining in lung macrophages and glomerular cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Bone marrow+3 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Tissue staining has low RNA concordance; verify independently (HPA tissue IHC) | |
| Regulation | Higher in breast tumors; inverse to ESR1 (UniProt) | |
| Isoform / epitope | No isoforms or precursor cleavage reported (UniProt) |
Section 1
Recommended CUEDC2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol appears alongside published CUEDC2 protocols for colorectal cancer, rat brain, and serous ovarian cancer (PMC6381460; PMC7906146; PMC7521062).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet PA1962); 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 Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear 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-CUEDC2, 0.5-1μg/ml (datasheet PA1962) |
| 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 | CUEDC2-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in a few tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page retrieval rule: nuclear antigen); colorectal and ovarian protocols used citrate pH 6.0 (PMC6381460; PMC7521062).
Section 2
What Is the Expected CUEDC2 Staining Pattern?
In paraffin-section IHC, expect CUEDC2 staining mainly in the cytoplasm of selected cells, including bone marrow hematopoietic cells, glomerular cells, lung macrophages and smooth muscle cells (HPA tissue IHC: Medium; cytoplasmic expression in a few tissues). Nuclear staining is biologically plausible (UniProt Q9H467: cytoplasm and nucleus). CUEDC2 has no transmembrane segment (UniProt Q9H467 topology). Treat the tissue pattern as provisional: HPA rates its IHC reliability Uncertain because staining and RNA expression show low consistency.
| Cytoplasmic staining in lung macrophages or smooth muscle cells, with other cells less conspicuous. | This matches reported Medium staining in those cell populations (HPA tissue IHC). Compare the cell pattern with a control section; HPA calls the tissue IHC profile Uncertain, so agreement alone does not establish antibody specificity. |
| Nuclear staining accompanies cytoplasmic staining in a plausible positive cell population. | Nuclear localization is consistent with UniProt Q9H467 and supported nucleoplasmic ICC-IF localization (HPA subcellular). HPA describes tissue IHC chiefly as cytoplasmic; assess prominent nuclear IHC against controls before calling it a confirmed tissue pattern. |
| Strong extracellular or luminal deposits dominate while expected intracellular staining is absent. | That distribution does not fit the reported cytoplasmic and nuclear localization (UniProt Q9H467; HPA tissue IHC). Consider precipitated chromogen, trapped reagent or nonspecific staining; inspect the no-primary control and tissue morphology before interpreting it as CUEDC2. |
| Adipocytes or bronchial respiratory epithelial cells stain strongly. | Both populations are reported as Not detected in the listed HPA tissue IHC observations. Suspect cross-reactivity or endogenous detection activity, then check matched controls. An HPA negative observation is a reference pattern, not proof that every specimen must be negative. |
| No intracellular signal appears in a section containing an expected positive cell population. | Bone marrow hematopoietic cells, glomerular cells, lung macrophages and smooth muscle cells each have reported Medium staining (HPA tissue IHC). Check cell identity and controls first; the profile's Uncertain reliability prevents treating any one section as an infallible positive control. |
💡Expected CUEDC2 appearanceCall a result plausible when identifiable HPA-listed positive cells show Medium, mainly cytoplasmic staining, with possible nuclear signal (HPA tissue IHC; UniProt Q9H467); dominant extracellular deposits or strong staining of HPA-listed negative cells warrant investigation.
| Which cells provide a provisional positive reference? | Bone marrow hematopoietic cells, kidney glomerular cells, lung macrophages and smooth muscle cells show Medium staining (HPA tissue IHC). Confirm that the relevant cells are present in the actual section before judging assay sensitivity. |
| How much confidence does the tissue IHC map carry? | HPA rates it Uncertain: antibody staining and RNA expression have low consistency, with external verification pending (HPA tissue IHC). HPA036544 also has an Uncertain IHC validation status (HPA antibodies). Interpret distribution alongside controls. |
| Does protein architecture predict membrane staining or an epitope effect? | CUEDC2 lacks a transmembrane segment, has no reported signal peptide or propeptide, and contains a CUE domain at residues 144–187 (UniProt Q9H467). No antibody epitope or target-specific fixation effect is supplied; architecture cannot determine retrieval conditions. |
| IF/ICC Q&A: What localization should I expect on its separate guide page? | Main signals are nucleoplasmic and cytosolic; nuclear membrane localization is also supported, while plasma membrane and primary cilium tip assignments are uncertain (HPA subcellular). Both listed antibodies have Supported ICC status (HPA antibodies). These ICC-IF observations do not validate an IHC protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| A putative positive section has no signal. | The expected cells may be absent, or a general IHC step may have failed; HPA's positive pattern is provisional (HPA tissue IHC: Uncertain). | Locate the relevant cells on the counterstained section. As general IHC practice, verify the validated control, retrieval, primary incubation and detection reagents before changing conditions. |
| Most tissue compartments show a uniform brown haze. | Diffuse staining obscures the restricted cytoplasmic pattern reported by HPA (HPA tissue IHC). General IHC causes include excess primary or detection reagent and inadequate blocking or washing. | Compare a no-primary control; as general IHC practice, review blocking, washing and reagent concentration while retaining the same tissue and detection controls. |
| Adipocytes or bronchial respiratory epithelium stain strongly. | Those cells are listed as Not detected (HPA tissue IHC). Cross-reactivity or endogenous enzyme activity may explain an unexpected chromogenic signal. | Inspect a no-primary control and the detection-system controls. If signal persists, address endogenous activity using standard IHC controls; if primary-dependent, reassess antibody specificity. |
| Signal appears chiefly in lumina or extracellular deposits. | This conflicts with the intracellular localization record (UniProt Q9H467; HPA tissue IHC). Chromogen deposits or retained reagent are possible general IHC explanations. | Inspect section morphology and no-primary control; repeat the affected staining step with clean reagents if deposits are visible. Do not score deposits as positive cells. |
| Only intense nuclear staining appears in a tissue section. | Nuclear localization is plausible (UniProt Q9H467; HPA subcellular), but HPA's tissue IHC profile is mainly cytoplasmic and rated Uncertain. | Record nuclear and cytoplasmic compartments separately. Compare expected cell types and controls before assigning the nuclear-only IHC pattern to CUEDC2. |
| A tissue result disagrees with an ICC-IF image. | HPA supports nucleoplasm and cytosol in ICC-IF, while its tissue IHC profile is Uncertain (HPA subcellular; HPA tissue IHC). The preparations report different observations. | Interpret the paraffin section against tissue IHC controls and cell identity. Use ICC-IF localization as context, without treating it as validation of the chromogenic tissue result. |
Experiment evidence
IHC & IF images showing CUEDC2 staining
PA1962 · IHCAnti-CUEDC2 antibody, PA1962, IHC(P) IHC(P): Human Thyroid Cancer Tissue
A08885 · IFImmunocytochemistry of CUEDC2 in HeLa cells with CUEDC2 antibody at 5 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000107874-CUEDC2/tissue/cerebral+cortex#imid_9301752
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000107874-CUEDC2/tissue/colon#imid_9301686
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000107874-CUEDC2/tissue/liver#imid_9301695
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000107874-CUEDC2/tissue/kidney#imid_9301680
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036544; green = CUEDC2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036544; green = CUEDC2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CUEDC2 IHC & IF
🧪Run bone marrow first and score staining in hematopoietic cells, which HPA rates Medium (HPA: bone marrow, hematopoietic cells, Medium). Use adipose tissue as the negative tissue; its adipocytes are Not detected (HPA: adipose tissue, adipocytes, Not detected). If adipocytes are present on the bone marrow slide, treat them as candidate internal negative cells and expect only background staining, confirming their status against the tissue negative control (HPA: adipose tissue, adipocytes, Not detected).
Positive control tissue: Bone marrow (Hematopoietic cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CUEDC2 in A-431, U-251MG, U2OS, HEK293, ASC52telo, RPTEC/TERT1, KOLF2.1J, NIH 3T3, with annotated localisation: Nucleoplasm (supported), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a host-species- and immunoglobulin-class-matched isotype control for a monoclonal primary, or matched nonimmune serum for a polyclonal primary; and CUEDC2-knockout material as a biological negative. In bone marrow, block endogenous peroxidase and check for residual signal in the no-primary control because hematopoietic cells can contribute peroxidase background (HPA: bone marrow, hematopoietic cells, Medium).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence; the selected PA1962 paraffin-section caption does not state a fixative (selected PA1962 tissue-IHC caption: fixative not stated). A CUEDC2-specific antigen-retrieval requirement and any advantage of frozen sections or IF are unreported; optimize retrieval against the bone marrow positive and adipose negative controls (HPA: bone marrow, hematopoietic cells, Medium; adipose tissue, adipocytes, Not detected). IF can assess the supported nucleoplasmic and cytosolic locations, while peroxidase background is a practical concern for chromogenic IHC in bone marrow (HPA subcellular: nucleoplasm and cytosol supported; HPA: bone marrow, hematopoietic cells, Medium).
HPA tissue IHC evidence for CUEDC2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. 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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | Medium | Protein (IHC) | HPA → |
| Lung | Macrophages | Medium | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CUEDC2 IHC Tips
Troubleshoot CUEDC2 staining in paraffin sections by checking retrieval, cellular compartment, controls and scoring before interpreting a chromogenic signal.
What retrieval should I try first if CUEDC2 staining is weak?
Start with Tris-EDTA at pH 9.0, heated to 95–98 °C for 20 min (page retrieval rule: nuclear antigen). Allow slides to cool in the retrieval buffer, then compare a positive control and a no-primary control processed in the same run (standard IHC practice). Assess both cytoplasmic and nuclear staining, since CUEDC2 is reported in both compartments (UniProt Q9H467: subcellular location). If signal remains weak, vary heating time on serial sections while keeping detection and imaging conditions fixed, and reject conditions that increase diffuse background or damage tissue morphology (standard IHC practice).
How should I investigate weak CUEDC2 staining after fixation?
CUEDC2-specific sensitivity to fixative type or fixation duration is unknown from the supplied evidence (selected PA1962 caption: fixative not stated). Record the fixative, time to fixation, fixation duration and section age for each specimen so weak staining can be compared with handling history (standard IHC practice). Run similarly processed positive-control tissue beside the specimen, using lung macrophages or kidney glomerular cells as candidates while recognizing that their reported staining is only medium and uncertain (HPA tissue IHC: medium staining; reliability uncertain). If staining varies with processing, compare matched serial sections and optimize retrieval systematically before attributing the difference to CUEDC2 biology (standard IHC practice).
Should CUEDC2 appear in nuclei, cytoplasm, or both?
Evaluate nuclear and cytoplasmic compartments separately because CUEDC2 is reported in both (UniProt Q9H467: subcellular location). Nucleoplasm and cytosol are supported locations in cell imaging, whereas plasma membrane and primary cilium tip assignments are uncertain (HPA subcellular: main and additional locations). The protein has no transmembrane segment, so a crisp membrane-only rim warrants closer review rather than automatic acceptance as CUEDC2 staining (UniProt Q9H467: topology; standard IHC interpretation). Compare the pattern with a no-primary slide and tissue morphology, and document compartment-specific intensity instead of combining all brown signal into one score (standard IHC practice).
Could epitope accessibility explain inconsistent CUEDC2 staining?
The supplied record lists a single 1–287 chain, a CUE domain at residues 144–187 and no annotated isoforms (UniProt Q9H467: processing, domains and isoforms). It also lists phosphoserine at residue 110, but gives no antibody epitope, so a phosphorylation-dependent staining effect cannot be inferred (UniProt Q9H467: modified residues; supplied antibody evidence: epitope unspecified). Check the antibody's documented immunogen or epitope before interpreting loss of signal as a change in CUEDC2 abundance (standard IHC practice). If staining changes after retrieval adjustments, compare serial sections with constant antibody and detection conditions, then report the observed staining change without assigning it to a particular domain or modification (standard IHC practice).
How can IF help check an ambiguous CUEDC2 IHC pattern?
Use IF as a separate localisation check, pairing CUEDC2 with a marker for the cell population being assessed, such as a macrophage marker when examining lung macrophages (HPA tissue IHC: lung macrophages, medium staining; standard IF practice). Choose spectrally separated fluorophores and favor a far-red CUEDC2 channel when tissue autofluorescence obscures shorter wavelengths (standard IF practice). Because the expected signals include nucleoplasm and cytosol, optimize permeabilisation for intracellular access, for example by comparing brief 0.1% Triton X-100 treatment with a gentler condition (HPA subcellular: supported locations; standard IF practice). Include single-label and no-primary controls before comparing IF localisation with chromogenic IHC (standard IF practice).
What should I check when CUEDC2 staining looks diffuse?
Compare the stained slide with no-primary and detection-only controls to identify nonspecific reagent signal (standard IHC practice). In chromogenic detection, block endogenous peroxidase before applying the detection reagent and inspect whether brown signal follows tissue structures or appears as a uniform haze (standard IHC practice). Titrate the primary antibody and shorten chromogen development on serial sections if background rises with staining strength; keep retrieval unchanged during that comparison (standard IHC practice). Judge any remaining signal against the reported cytoplasmic tissue pattern and supported nuclear and cytosolic locations, while treating the tissue IHC reference cautiously because its reliability is uncertain (HPA tissue IHC: profile and reliability; HPA subcellular: main locations).
How should I score CUEDC2 across heterogeneous paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, then record nuclear and cytoplasmic staining separately (UniProt Q9H467: subcellular location; standard IHC practice). For a chosen compartment, report the percentage of positive cells and an H-score from intensity categories 0–3; for sparse positive cells, density per mm² can be useful (standard IHC quantification practice). Normalize counts to viable cells or measured viable tissue area, and apply the same threshold, region selection and detection settings across sections (standard IHC quantification practice). Keep tumor and adjacent normal regions distinct when testing the reported breast tumor association, and avoid treating an HPA medium-staining tissue as a calibrated abundance standard (UniProt Q9H467: tissue specificity; HPA tissue IHC: reliability uncertain).
How do I distinguish plausible CUEDC2 staining from artefact?
Give greatest weight to reproducible nuclear or cytoplasmic signal in intact cells, the reported CUEDC2 compartments (UniProt Q9H467: subcellular location; HPA subcellular: supported locations). Review membrane-only signal, staining confined to section edges or necrotic regions, and brown deposits that persist in no-primary controls as possible artefacts (UniProt Q9H467: no transmembrane segment; standard IHC interpretation). Check cell identity: lung macrophages and kidney glomerular cells have reported medium staining, although the tissue IHC dataset has uncertain reliability (HPA tissue IHC: positive cells and reliability). The PA1962 thyroid cancer image demonstrates tissue staining, but its caption does not state a fixative; it cannot establish an FFPE-specific pattern or validate an unexpected compartment (selected PA1962 tissue-IHC caption).
Boster reagents
Best CUEDC2 / CUE domain-containing protein 2 IHC Antibodies
CUEDC2 reagents have illustrated paraffin-section IHC in human thyroid cancer tissue (PA1962 image caption) and ICC in HeLa cells (A08885 image caption); both list Human, Mouse and Rat reactivity (catalog).
Real IHC data
Anti-CUEDC2 Antibody ®
Cat # PA1962
Real IF data
Anti-CUEDC2 Antibody
Cat # A08885
PA1962 lists IHC and shows staining of human thyroid cancer tissue in a paraffin section (catalog applications; PA1962 image caption). A08885 lists ICC and shows staining of HeLa cells at 5 μg/mL (catalog applications; A08885 image caption).
Which to pick: Choose PA1962 for paraffin-section IHC: its image caption documents IHC(P) in human thyroid cancer tissue; the fixative is unreported (PA1962 image caption). Choose A08885 for cultured-cell IF/ICC: its ICC image shows HeLa cells at 5 μg/mL (A08885 image caption). Both list Human, Mouse and Rat reactivity, so the catalog gives neither a cross-species advantage; both are rabbit antibodies with clone type unreported (catalog reactivity, host and clone fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9H467 (CUED2_HUMAN, CUE domain-containing protein 2).
- Human Protein Atlas. CUEDC2 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. CUEDC2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and cytosol. In addition localized to the nuclear membrane, plasma membrane and primary cilium tip..
- Human Protein Atlas. CUEDC2 antibody validation summary (2 antibodies).
- Expression of CUEDC2 in colorectal cancer with different invasion and migration abilities. The Journal of international medical research 2019 — PMC6381460.
- CUEDC2 ablation enhances the efficacy of mesenchymal stem cells in ameliorating cerebral ischemia/reperfusion insult. Aging 2021 — PMC7906146.
- CUEDC2 modulates cardiomyocyte oxidative capacity by regulating GPX1 stability. EMBO molecular medicine 2016 — PMC4931293.
- Expression of CUE domain containing 2 protein in serous ovarian cancer tissue: predicting disease-free and overall survival of patients. The Journal of international medical research 2020 — PMC7521062.
- PubMed PMID:15164054 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:17347654 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CUEDC2 (CUE domain-containing protein 2). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); nuclear location annotated (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: PA1962 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet PA1962); 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 Tris-EDTA buffer, pH 9.0 , 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-CUEDC2, 0.5-1μg/ml (datasheet PA1962); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Tissue staining has low RNA concordance; verify independently (HPA tissue IHC); Isoform / epitope: No isoforms or precursor cleavage reported (UniProt); Regulation: Higher in breast tumors; inverse to ESR1 (UniProt). Evidence: UniProt Q9H467, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the nucleoplasm and cytosol. In addition…, Datasheet PA1962, PMC6381460, PMC7906146. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CUEDC2/immunohistochemistry-ihc-guide.