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CUEDC2 / CUE domain-containing protein 2 · IHC design guide
Design Immunohistochemistry for CUEDC2
Plan chromogenic paraffin IHC for CUEDC2 using the IHC-validated antibody at 0.5–1 μg/mL (datasheet: PA1962). Assess cytoplasmic staining in lung macrophages and kidney glomerular cells, interpreting results cautiously because tissue staining has low antibody–RNA consistency (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 formalin fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Tissue staining has low antibody–RNA consistency (HPA tissue IHC) | |
| Regulation | Higher in high-grade breast tumors (UniProt) | |
| Isoform / epitope | 0 annotated isoforms; one 1–287 chain (UniProt) |
Section 1
Recommended CUEDC2 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with three published CUEDC2 tissue-staining workflows (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 the page’s Tris-EDTA pH 9.0, 95–98 °C, 20 min retrieval; the two detailed published workflows used citrate pH 6.0 (page retrieval rule; PMC6381460; PMC7521062).
Section 2
What Is the Expected CUEDC2 Staining Pattern?
CUEDC2 is found in the cytoplasm and nucleus (UniProt Q9H467). In tissue IHC, HPA reports cytoplasmic staining at medium levels in lung macrophages, bone marrow hematopoietic cells, kidney glomerular cells, and smooth muscle cells (HPA tissue IHC). CUEDC2 has no transmembrane segment or signal peptide (UniProt Q9H467 topology). Use these cell patterns as provisional references: HPA rates its tissue IHC profile Uncertain because staining and RNA expression have low consistency (HPA tissue IHC).
| Brown cytoplasmic staining in lung macrophages or smooth muscle cells. | This fits the reported medium tissue IHC pattern (HPA tissue IHC). Score the named cells separately from neighboring cells (general IHC practice). The pattern is a provisional match because tissue IHC reliability is Uncertain (HPA tissue IHC). |
| Dominant extracellular or luminal staining with little signal in the named cells. | Treat this as a possible staining artefact: CUEDC2 has no signal peptide or transmembrane segment (UniProt Q9H467 topology), while HPA describes a cytoplasmic tissue pattern (HPA tissue IHC). Check whether the signal persists in a no-primary control (general IHC practice). |
| Strong staining in a cell population reported as not detected. | For example, HPA reports adipocytes as Not detected (HPA tissue IHC). Investigate antibody cross-reactivity or endogenous detection activity with controls (general IHC practice). HPA's Uncertain IHC rating means the discrepancy alone cannot prove a false positive (HPA tissue IHC). |
| Diffuse brown haze across cells and surrounding tissue. | This does not resolve the cell-restricted cytoplasmic pattern reported by HPA (HPA tissue IHC). Check the no-primary control and inspect washing, blocking, and chromogen development for background staining (general IHC practice). |
| No signal in a section containing lung macrophages. | HPA reports medium staining in those cells, but its tissue IHC profile is Uncertain (HPA tissue IHC). Confirm that the expected cells are present, then review detection, antigen retrieval, and antibody conditions (general IHC practice). |
💡Expected CUEDC2 appearanceCall a provisional positive when identifiable lung macrophages, bone marrow hematopoietic cells, glomerular cells, or smooth muscle cells show medium cytoplasmic staining (HPA tissue IHC); dominant extracellular signal or staining spread across unrelated cells warrants a control check (UniProt Q9H467 topology; general IHC practice).
| Tissue reference strength | HPA reports medium staining in four named cell populations but rates the tissue profile Uncertain for low staining–RNA consistency (HPA tissue IHC). Compare cell identity and compartment before interpreting intensity. |
| Compartment and topology | Cytoplasm and nucleus are listed by UniProt; HPA tissue IHC describes cytoplasmic expression (UniProt Q9H467; HPA tissue IHC). CUEDC2 has no signal peptide or transmembrane segment (UniProt Q9H467 topology). |
| Breast tumor context | Protein levels are reported higher in breast tumors than matched adjacent normal tissue and lower in low-grade than high-grade tumors (UniProt Q9H467). This comparison does not set a staining cutoff for an individual section. |
| Antibody validation | HPA036544 is Uncertain for IHC and Supported for ICC; HPA041829 has no IHC status listed and is Supported for ICC (HPA antibodies). ICC support does not establish an IHC staining pattern. |
| IF/ICC Q: Should nuclear signal count? | A: Nucleoplasm and cytosol are Supported ICC-IF locations (HPA subcellular). Interpret IF/ICC on its own guide page; HPA tissue IHC reports a cytoplasmic profile with Uncertain reliability (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected cells are absent from the section. | The chosen field cannot assess the named HPA cell population (HPA tissue IHC). | Locate a field containing identifiable macrophages, hematopoietic cells, glomerular cells, or smooth muscle cells before scoring (HPA tissue IHC; general IHC practice). |
| Named cells are present but entirely unstained. | The reported medium signal may be missed, or staining may have failed; HPA IHC reliability is Uncertain (HPA tissue IHC). | Check a working control section, antibody conditions, retrieval, and detection reagents in sequence (general IHC practice). |
| Extracellular or luminal material stains prominently. | That distribution conflicts with the reported cytoplasmic tissue pattern (HPA tissue IHC) and CUEDC2 topology (UniProt Q9H467). | Compare a no-primary control and assess nonspecific binding or deposited chromogen before assigning CUEDC2 positivity (general IHC practice). |
| Many unrelated cells show uniform brown staining. | Broad signal is difficult to reconcile with HPA's cytoplasmic staining in a few tissues (HPA tissue IHC); background or cross-reactivity is possible (general IHC practice). | Inspect a no-primary control, then reassess blocking, antibody concentration, washing, and cell identity (general IHC practice). |
| A nuclear-only IHC pattern appears. | Nuclear localization is possible (UniProt Q9H467), but HPA's tissue IHC summary emphasizes cytoplasm (HPA tissue IHC). | Record nuclear and cytoplasmic scores separately; compare control sections before calling the nuclear pattern specific (general IHC practice). |
| A proposed negative control stains. | HPA lists some populations as Not detected, including adipocytes, but its IHC profile remains Uncertain (HPA tissue IHC). | Check cell identification and no-primary signal, then verify the result with an independent antibody or orthogonal evidence where available (general IHC practice). |
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 lung first: macrophages should stain (HPA: Medium in lung macrophages). Use adipose tissue as the negative, expecting no adipocyte staining (HPA: Not detected in adipocytes); on the lung slide, treat neighboring non-macrophage cells as internal negatives only where they lack specific signal.
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 primary-host- and immunoglobulin-class-matched isotype control (or host-matched nonimmune IgG for a polyclonal primary), and CUEDC2 knockout material or validated immunizing-peptide competition as a biological specificity control (standard IHC practice). Quench endogenous peroxidase and check for pigment in lung macrophages during chromogenic detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported in the supplied evidence, and the selected PA1962 IHC(P) caption does not report a fixative (PA1962 caption). Retrieval dependence is unreported; optimize antigen retrieval using the lung positive and adipose negative controls (HPA: Medium in lung macrophages; Not detected in adipocytes). The evidence does not establish whether frozen sections or IF/ICC are easier; IF/ICC scoring can use the supported nucleoplasm and cytosol locations, with a no-primary control to assess lung macrophage autofluorescence (HPA subcellular: supported nucleoplasm and cytosol; standard IF practice).
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
These answers address CUEDC2 staining in chromogenic paraffin-section IHC, with one entry on adapting the experiment to IF/ICC.
How should I optimize retrieval when 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). Keep section thickness, cooling, antibody incubation, and chromogen development constant while comparing retrieved sections with a no-retrieval section (standard IHC practice). Score nuclear and cytoplasmic staining separately, because both compartments are plausible for CUEDC2 (UniProt: nucleus and cytoplasm; HPA subcellular: supported nucleoplasm and cytosol). If staining remains weak or tissue morphology suffers, test a milder retrieval condition as a fallback and retain the condition that improves interpretable cellular signal without raising background (standard IHC practice).
Could fixation explain weak or uneven CUEDC2 staining?
CUEDC2-specific sensitivity to fixation is unknown from the supplied evidence (selected-SKU caption: fixative unreported). For a troubleshooting comparison, process matched samples with documented fixation and processing histories, then stain sections together using the same retrieval, antibody incubation, and detection steps (standard IHC practice). Assess tissue preservation and staining in both nuclear and cytoplasmic compartments before attributing a difference to antigen loss (UniProt: nucleus and cytoplasm; standard IHC practice). Record fixation conditions alongside the results, because uneven penetration or processing can create spatial staining differences in paraffin sections (standard IHC practice); the thyroid-cancer IHC caption alone cannot establish a CUEDC2-specific fixation preference (PA1962 caption: fixative unreported).
Should CUEDC2 staining be nuclear, cytoplasmic, or both?
Evaluate nuclear and cytoplasmic signal separately in each identifiable cell population (UniProt: nucleus and cytoplasm; standard IHC practice). Nucleoplasm and cytosol are supported locations, while nuclear membrane staining is additional; plasma membrane and primary-cilium-tip locations carry uncertain support (HPA subcellular: main and additional locations). CUEDC2 has no transmembrane segment, so isolated crisp membrane staining warrants a check for nonspecific signal rather than an automatic positive call (UniProt: topology; standard IHC interpretation). Compare serial sections with and without primary antibody, inspect cell morphology, and document which compartment contributes to any score (standard IHC practice); a compartment pattern alone does not establish antibody specificity (standard IHC interpretation).
How can epitope accessibility complicate CUEDC2 IHC interpretation?
The supplied record lists 0 isoforms and a CUE domain at residues 144–187, but gives no antibody epitope, so a sequence-specific retrieval claim is unsupported (UniProt: isoforms and domain; supplied antibody evidence: epitope unreported). CUEDC2 also has a reported phosphoserine at residue 110; the evidence does not establish whether this modification affects this antibody’s staining (UniProt: modified residue; supplied antibody evidence: epitope unreported). Compare retrieval conditions on adjacent sections while holding detection constant, and assess morphology alongside signal (standard IHC practice). If discordant patterns persist, seek an independently characterized antibody with a documented epitope before assigning the discrepancy to an isoform or modification (standard IHC validation practice).
How would I adapt CUEDC2 localisation checks to IF/ICC?
Plan IF/ICC as a separately optimized experiment: the selected antibody’s caption documents tissue IHC(P), while HPA reports IF/ICC images for CUEDC2 in listed cell lines (PA1962 caption; HPA subcellular: ICC/IF images). Multiplex CUEDC2 with an independently validated marker for the cell population being examined, and use a nuclear counterstain to distinguish nucleoplasm from cytosol (HPA subcellular: supported locations; standard IF practice). Choose fluorophores after checking the specimen’s autofluorescence and include single-channel controls for bleed-through (standard IF practice). Because CUEDC2 has no transmembrane segment, optimize permeabilisation to reach intracellular epitopes while preserving nuclear and cytosolic morphology; the antibody’s exact epitope and IF fixation sensitivity are unreported (UniProt: topology; supplied antibody evidence: epitope and IF fixation unreported).
What should I check when CUEDC2 sections show widespread brown staining?
Run a section without primary antibody and inspect whether staining persists after the usual peroxidase block; persistent signal points to detection-system or endogenous-enzyme background (standard chromogenic IHC practice). Compare tissue edges, folds, and damaged areas with well-preserved interior cells, since staining concentrated at artifacts can mislead scoring (standard IHC practice). Check whether longer chromogen development increases diffuse signal without improving nuclear or cytoplasmic definition (UniProt: nucleus and cytoplasm; standard IHC practice). Optimize blocking, primary-antibody concentration, washes, and development one variable at a time, recording the effect on cellular contrast (standard IHC practice); HPA’s tissue-IHC reliability is uncertain, so its staining pattern is a cautious comparator (HPA tissue IHC: uncertain reliability).
How should I score heterogeneous CUEDC2 staining across a section? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and scoring region before reviewing stain intensity, and exclude folds, edges, and necrotic areas by a consistent rule (standard IHC quantification practice). Report an H-score from intensity-weighted percentages or report the percentage of positive cells, with nuclear and cytoplasmic scores kept separate (UniProt: nucleus and cytoplasm; standard IHC quantification practice). For discrete positive cells, density per mm² can supplement these scores; normalize counts to the viable tissue area or the number of eligible cells actually assessed (standard IHC quantification practice). Use the same threshold, exposure or scan settings, and analysis rules across samples, and state which cell type contributed to each result (standard IHC quantification practice).
How can I distinguish genuine CUEDC2 positivity from staining artifacts?
Favor reproducible intracellular staining with identifiable nuclear or cytoplasmic boundaries, because these are supported CUEDC2 compartments (UniProt: nucleus and cytoplasm; HPA subcellular: supported nucleoplasm and cytosol). Check the stained cell population rather than treating every brown area as equivalent: HPA reports medium staining in lung macrophages and kidney glomerular cells, but rates its tissue-IHC reliability uncertain (HPA tissue IHC: positive cells and uncertain reliability). Isolated membrane-only signal, section-edge accentuation, necrotic deposits, or signal in the no-primary control requires further investigation (UniProt: no transmembrane segment; standard IHC interpretation). Review morphology, peroxidase-block controls, and serial sections before interpreting a tumor-versus-adjacent difference (standard IHC practice; UniProt: reported breast-tumor comparison).
Boster reagents
Best CUEDC2 / CUE domain-containing protein 2 IHC Antibodies
The catalog shows CUEDC2 IHC in human thyroid cancer paraffin sections (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 is listed for IHC and has an image of human thyroid cancer tissue in a paraffin section (catalog applications; PA1962 image caption). A08885 is listed for ICC and has an image of HeLa cells at 5 μg/mL (catalog applications; A08885 image caption).
Which to pick: Choose PA1962 for tissue IHC in paraffin sections based on its IHC application listing and its own tissue image (catalog applications; PA1962 image caption); the fixative is unreported (PA1962 image caption). Choose A08885 for IF/ICC in cultured cells based on its ICC application listing and HeLa cell image (catalog applications; A08885 image caption). For mouse or rat samples, both list reactivity with those species, while the supplied IHC and ICC images document human samples only (catalog reactivity; PA1962 and A08885 image captions).
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 formalin fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Tissue staining has low antibody–RNA consistency (HPA tissue IHC); Isoform / epitope: 0 annotated isoforms; one 1–287 chain (UniProt); Regulation: Higher in high-grade breast tumors (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.