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ODF2L / Protein BCAP · IHC design guide
Design Immunohistochemistry for ODF2L
Plan chromogenic IHC for ODF2L in paraffin sections using the catalog antibody’s 2–5 μg/ml range (datasheet A16444-1). Assess cytoplasmic staining against the HPA tissue profile, which awaits external verification (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ODF2L 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 stain (HPA tissue IHC); centrioles and basal bodies are molecular locations (UniProt) | |
| Staining pattern | Cytoplasmic staining in glandular and respiratory epithelial cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A16444-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Cervix+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 awaits external verification (HPA tissue IHC) | |
| Regulation | Lost during ciliogenesis; returns at lower levels (UniProt) | |
| Isoform / epitope | Six isoforms; antibody epitope coverage is unspecified (UniProt) |
Section 1
Recommended ODF2L IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A16444-1) and a published ODF2L lung-tissue IHC protocol (PMC13494581) provide starting conditions.
| Sample | Paraffin-embedded human testicular seminoma tissue; fixative not specified (datasheet A16444-1) |
| Fixation | Image fixative and duration unreported (datasheet A16444-1); 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 A16444-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A16444-1) |
| Primary antibody | Rabbit anti-ODF2L, 2-5 μg/ml (datasheet A16444-1) |
| Primary incubation | Overnight at 4 °C (datasheet A16444-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A16444-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ODF2L-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet: A16444-1); the published protocol used EDTA without specifying pH (PMC13494581).
Section 2
What Is the Expected ODF2L Staining Pattern?
ODF2L localizes to centrosomes, centrioles, centriolar satellites and ciliary basal bodies; it has no transmembrane segment (UniProt Q9ULJ1). In paraffin IHC, expect predominantly cytoplasmic staining in cell populations reported as positive, including bronchial respiratory epithelium and glandular cells in several tissues (HPA tissue IHC). Treat intensity as provisional: HPA calls the tissue pattern “Uncertain,” pending external verification (HPA tissue IHC).
| Cytoplasmic stain in bronchial respiratory epithelial cells or appendix, colon or duodenal glandular cells (HPA tissue IHC). | This fits the reported medium staining in those populations (HPA tissue IHC). Centrosomal or basal-body enrichment would fit the annotated localization, but a resolved punctum is not required in chromogenic tissue sections (UniProt Q9ULJ1; general IHC practice). |
| Predominantly nuclear, membrane-rim or extracellular staining, without a convincing cytoplasmic component. | That distribution does not match the reported general cytoplasmic IHC profile or the annotated cytoskeletal locations (HPA tissue IHC; UniProt Q9ULJ1). Check morphology and controls before treating it as ODF2L; chromogen deposits or nonspecific binding can mislead compartment assignment (general IHC practice). |
| Strong staining in a cell population reported as undetected, such as lymph-node germinal-center cells (HPA tissue IHC). | Consider cross-reactivity or endogenous detection activity, and compare a no-primary control (general IHC practice). HPA’s undetected call is a reference observation, not proof that every specimen must be negative; its IHC reliability is “Uncertain” (HPA tissue IHC). |
| Diffuse color across tissue and empty spaces, with poor separation between cells. | This is difficult to score as cellular staining (general IHC practice). Review the no-primary control and staining workflow for background from detection reagents, blocking or washing; do not infer ODF2L expression from diffuse color alone (general IHC practice). |
| No signal in bronchial respiratory epithelium or another HPA-reported medium-staining population (HPA tissue IHC). | First confirm that the expected cells are present and the detection run worked (general IHC practice). An absent signal warrants a controlled repeat, but HPA’s uncertain reliability prevents treating one negative section as a definitive biological contradiction (HPA tissue IHC). |
💡Expected ODF2L appearanceCall a positive result when cell-associated cytoplasmic staining is discernible in an HPA-reported positive population, such as medium bronchial respiratory epithelium; isolated nuclear, membrane-rim or diffuse extracellular color is suspect (HPA tissue IHC; UniProt Q9ULJ1; general IHC practice).
| Tissue and cell population | HPA reports medium staining in bronchial respiratory epithelium and several glandular populations, but no staining in lymph-node germinal-center cells; score identified cells rather than whole-section color (HPA tissue IHC; general IHC practice). |
| Ciliary state and optical resolution | UniProt places ODF2L at centrioles in proliferating cells and basal bodies in ciliated cells, with levels falling during ciliogenesis and returning at lower levels afterward (UniProt Q9ULJ1). Routine chromogenic IHC may show cytoplasm without resolving those structures (general IHC practice). |
| Isoforms and antibody validation | Six isoforms are listed, but the supplied record gives no antibody epitope or isoform coverage (UniProt Q9ULJ1). HPA rates IHC for HPA028020, HPA028095 and HPA028333 “Uncertain”; apparent positives need independent support (HPA antibodies). |
| IHC detection background | Endogenous enzyme activity or nonspecific detection can produce color unrelated to the primary antibody (general IHC practice). A no-primary control helps identify detection background, while tissue morphology and localization help assess the remaining stain (general IHC practice). |
| IF/ICC Q: Is a punctate pattern established? | A: Centrosomal and basal-body puncta are biologically plausible (UniProt Q9ULJ1), but HPA supplies no ICC-IF images or assigned main subcellular location for ODF2L; an IF pattern is therefore not independently established here (HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected cytoplasmic signal is absent in bronchial respiratory epithelium (HPA tissue IHC). | The relevant cells may be missing, or an IHC workflow step may have failed (general IHC practice). | Verify cell identity, compare a concurrent positive tissue section, and review retrieval, primary incubation and detection against the catalog antibody’s IHC-P instructions (general IHC practice). |
| All tissue and control areas show similar diffuse color. | Detection background or inadequate washing may obscure cellular staining (general IHC practice). | Inspect a no-primary control; review blocking, washes and detection-reagent exposure, then score only cell-associated stain above background (general IHC practice). |
| The dominant signal is nuclear or follows cell borders. | The compartment conflicts with HPA’s general cytoplasmic profile and UniProt’s centrosomal and basal-body annotations (HPA tissue IHC; UniProt Q9ULJ1). | Recheck focus and cell boundaries, compare no-primary staining, and seek an independently supported antibody result before assigning ODF2L localization (general IHC practice; HPA antibodies). |
| Germinal-center cells stain strongly despite HPA reporting no detection there (HPA tissue IHC). | Cross-reactivity or endogenous detection activity is possible; HPA tissue reliability is “Uncertain” (general IHC practice; HPA tissue IHC). | Compare no-primary and independent-antibody controls where available, and document the cell population and staining compartment before interpreting the discrepancy (general IHC practice). |
| Cytoplasmic stain appears, but no discrete basal-body or centrosomal dots can be resolved. | Those locations are annotated, while chromogenic sections may not resolve small intracellular structures (UniProt Q9ULJ1; general IHC practice). | Assess the cytoplasmic pattern and cell identity in IHC; consult the separate IF/ICC guide if subcellular resolution is required (HPA tissue IHC; general IHC practice). |
| Two antibodies give different tissue patterns. | The supplied antibodies have uncertain IHC validation, and their epitopes and isoform coverage are unspecified (HPA antibodies; UniProt Q9ULJ1). | Compare staining in the same defined cell populations with matched controls; report the disagreement rather than choosing a pattern solely by intensity (general IHC practice; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing ODF2L staining
A16444-1 · IHCIHC analysis of ODF2L using anti-ODF2L antibody (A16444-1). ODF2L was detected in a paraffin-embedded section of human testicular seminoma t
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000122417-ODF2L/tissue/cerebral+cortex#imid_7807124
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000122417-ODF2L/tissue/colon#imid_7807058
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000122417-ODF2L/tissue/liver#imid_7807059
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000122417-ODF2L/tissue/kidney#imid_7807060
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ODF2L IHC & IF
🧪Run bronchus first and expect staining in respiratory epithelial cells (HPA: Medium in bronchus respiratory epithelial cells). Use cervix glandular cells as the negative tissue (HPA: Not detected in cervix glandular cells); on the bronchus slide, cells without specific signal should show only background staining, without assuming a particular neighboring cell type is negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Cervix (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for ODF2L; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; a concentration-matched rabbit IgG isotype control (selected-SKU caption: rabbit primary antibody); and a biological negative such as ODF2L knockout tissue or a validated immunizing-peptide block. For chromogenic bronchus sections, quench endogenous peroxidase and inspect inflammatory cells for residual background before scoring epithelial staining.
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU tissue-IHC caption: fixative not stated). The demonstrated IHC conditions use heat retrieval in EDTA at pH 8.0, but the evidence does not establish that retrieval is required or that frozen sections or IF are easier (selected-SKU tissue-IHC caption: EDTA retrieval; HPA subcellular: no ICC-IF image cell lines). In bronchus, check inflammatory-cell peroxidase background when interpreting respiratory epithelial staining (HPA: Medium in bronchus respiratory epithelial cells).
HPA tissue IHC evidence for ODF2L
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Chondrocytes | Not detected | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Not detected | Protein (IHC) | HPA → |
| Vagina | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ODF2L IHC Tips
Troubleshoot ODF2L chromogenic IHC in paraffin sections using the catalog antibody’s tissue image, the reported localisation, and appropriately qualified tissue controls.
Which retrieval condition should I start with for ODF2L staining in paraffin sections?
For paraffin-section chromogenic IHC, begin with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A16444-1). That condition accompanied ODF2L staining in a human testicular seminoma section, although the heating time was not reported (datasheet A16444-1). Keep section thickness, heating, and cooling consistent across the test and control sections, then assess signal and tissue integrity together (standard IHC practice). If staining is weak, optimise heating duration before testing another buffer or pH as a fallback (standard IHC practice). Record any departure from the documented retrieval condition when comparing staining across runs.
How should I troubleshoot variable staining when the fixation history is uncertain?
ODF2L-specific sensitivity to fixation is unknown; the catalog antibody’s paraffin-section caption does not identify its fixative (datasheet A16444-1). Record the fixative and fixation duration for each specimen, and compare sections with similar processing histories where possible (standard IHC practice). Run a processing-matched control alongside the specimen before attributing a weak signal to low antigen abundance (standard IHC practice). If staining varies, change one processing or retrieval variable at a time while monitoring morphology and background (standard IHC practice). Neither the reported tissue pattern nor the protein’s localisation establishes a preferred fixation condition for ODF2L.
Where should convincing ODF2L staining appear within a positive cell?
Prioritise staining near centrioles or basal bodies when assessing cellular localisation: ODF2L is reported at centrioles in proliferative cells and basal bodies in ciliated cells (UniProt Q9ULJ1). Centriolar satellites are also reported locations, so a punctate cytoplasmic pattern can be biologically plausible (UniProt Q9ULJ1). Its abundance can fall during ciliogenesis and return at lower levels afterward, making ciliation state relevant to comparisons (UniProt Q9ULJ1). The tissue IHC profile describes general cytoplasmic expression but carries Uncertain reliability (HPA tissue IHC). Evaluate apparent diffuse DAB signal against morphology, a matched control, and the expected subcellular pattern before assigning it to ODF2L.
Could isoforms or an unknown epitope explain inconsistent staining?
ODF2L has 6 annotated isoforms, so isoform coverage matters when interpreting a negative section (UniProt Q9ULJ1). The supplied antibody caption does not identify its epitope or establish which isoforms it recognises (datasheet A16444-1). Ask for epitope mapping before claiming that one staining pattern represents every isoform, and retain the antibody identity in the experiment record. UniProt annotates no glycosylation sites or modified residues, but those annotations do not establish epitope accessibility after tissue processing (UniProt Q9ULJ1). Compare staining in consistently processed sections and avoid assigning an isoform from DAB localisation alone (standard IHC practice).
How can I assess ODF2L localisation by IF alongside chromogenic IHC?
For a separate IF experiment, multiplex ODF2L with an epithelial marker in bronchus to identify the expected respiratory epithelial cells (HPA tissue IHC: Medium in bronchus respiratory epithelial cells). Select a far-red channel where practical, and measure unstained tissue autofluorescence before interpreting puncta (standard IF practice). ODF2L has no annotated transmembrane segment and is reported at intracellular centrioles and basal bodies; use controlled permeabilisation to provide intracellular antibody access (UniProt Q9ULJ1). Titrate permeabilisation against structural preservation, because the antibody’s epitope is unspecified (datasheet A16444-1). The supplied HPA subcellular record lists no ICC/IF images, so confirm antibody performance in IF independently (HPA subcellular).
What should I change when DAB obscures a weak ODF2L signal?
First compare a no-primary section with the stained section to identify secondary-reagent or tissue-derived background (standard IHC practice). Include an endogenous peroxidase block before HRP and DAB detection, and examine whether residual colour follows the tissue rather than plausible cellular staining (standard IHC practice). The documented tissue run used 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A16444-1). Use that run as a starting reference, then optimise blocking, washing, and primary concentration while holding retrieval constant. Review tissue edges and damaged areas separately, since accumulated chromogen there can distort interpretation (standard IHC practice).
How should I score ODF2L staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the eligible cell population and compartment before scoring, then apply the same rule to every section (standard IHC practice). Record percentage positive cells and intensity on a 0–3 scale; their weighted sum gives an H-score from 0–300 (standard IHC practice). For spatial patterns, count positive cells per mm² of viable, evaluated tissue and report the evaluated area (standard IHC practice). Normalise comparisons to eligible cells or viable tissue area rather than whole-slide area when those denominators differ. Analyse centriolar or basal-body puncta separately from diffuse cytoplasmic colour, since both localisation reports and uncertain general cytoplasmic IHC exist (UniProt Q9ULJ1; HPA tissue IHC).
How can I distinguish genuine ODF2L staining from misleading positives?
A credible result should follow cell morphology and, where resolvable, the reported centriolar or basal-body localisation of ODF2L (UniProt Q9ULJ1). Treat isolated nuclear colour or staining confined to section edges and necrotic areas cautiously, and compare it with a no-primary control (standard IHC practice). Check persistent DAB colour against an endogenous peroxidase control before assigning it to ODF2L (standard IHC practice). Cell identity also matters: HPA reports medium staining in bone-marrow hematopoietic cells, whereas UniProt reports ODF2L as not detected in bone marrow (HPA tissue IHC; UniProt Q9ULJ1). This disagreement and HPA’s Uncertain IHC reliability warrant orthogonal confirmation before making tissue-specific claims (HPA tissue IHC).
Boster reagents
Best ODF2L / Protein BCAP IHC Antibodies
A16444-1 has real IHC data from a human paraffin-embedded testicular seminoma section (catalog image caption); no IF/ICC data are supplied (catalog applications and images).
Real IHC data
Anti-ODF2L Antibody ®
Cat # A16444-1
A16444-1 is listed for human IHC, with a paraffin-embedded testicular seminoma image (catalog applications, reactivity and image caption). Its listed applications do not include IF/ICC (catalog applications).
Which to pick: Choose A16444-1 for human paraffin-section IHC; its image caption reports EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody, while the catalog recommends 2–5 μg/ml for IHC (catalog image caption and dilution). The antibody is rabbit, its clonality is unreported, and the image caption does not report a fixative (catalog host, clone and image caption). No listed SKU supports an IF/ICC or cross-species recommendation because A16444-1 lists human reactivity and does not list IF/ICC (catalog reactivity and applications).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9ULJ1 (ODF2L_HUMAN, Protein BCAP).
- Human Protein Atlas. ODF2L tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. ODF2L subcellular location (ICC-IF): Highest expression in HUVEC/TERT2: 21.0 nTPM.
- Human Protein Atlas. ODF2L antibody validation summary (3 antibodies).
- SHAP-interpretable machine learning integrating exposures and multi-omics reveals immune alterations and biomarkers in COPD-lung cancer comorbidity. Frontiers in immunology 2026 — PMC13494581.
- PubMed PMID:10574462 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ODF2L (Protein BCAP). Expected localisation Cytoplasmic tissue stain (HPA tissue IHC); centrioles and basal bodies are molecular locations (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A16444-1). Validated IHC antibody: A16444-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Cervix. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human testicular seminoma tissue; fixative not specified (datasheet A16444-1); 2. Fixation: Image fixative and duration unreported (datasheet A16444-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A16444-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A16444-1); 8. Primary antibody: Rabbit anti-ODF2L, 2-5 μg/ml (datasheet A16444-1); 9. Primary incubation: Overnight at 4 °C (datasheet A16444-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Tissue staining awaits external verification (HPA tissue IHC); Isoform / epitope: Six isoforms; antibody epitope coverage is unspecified (UniProt); Regulation: Lost during ciliogenesis; returns at lower levels (UniProt). Evidence: UniProt Q9ULJ1, HPA tissue IHC · Uncertain, HPA cell-line RNA · Highest expression in HUVEC/TERT2: 21.0 nTPM, Datasheet A16444-1, PMC13494581. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ODF2L/immunohistochemistry-ihc-guide.