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DHX15 / ATP-dependent RNA helicase DHX15 · IHC design guide
Design Immunohistochemistry for DHX15
Plan DHX15 chromogenic IHC on paraffin sections using the nuclear and nucleolar tissue pattern (HPA tissue IHC). The guide covers catalog antibody conditions and tissue controls, with caution about low agreement between staining and RNA expression (datasheet A08140-1; HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DHX15 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 and nucleolar staining in several tissues (HPA tissue IHC) | |
| Staining pattern | Nuclear and nucleolar signal in several tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A08140-1) | |
| Positive control | Cerebellum+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 consistent across paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Cytoplasmic staining was observed and disregarded (HPA tissue IHC) | |
| Regulation | No expression regulator is specified (UniProt) | |
| Isoform / epitope | No annotated isoforms; one full-length chain (UniProt) |
Section 1
Recommended DHX15 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A08140-1) is followed by published DHX15 IHC methods from three articles (PMC6861564; PMC10894276; PMC8862312).
| Sample | Paraffin-embedded human prostate cancer tissue; fixative not specified (datasheet A08140-1) |
| Fixation | Image fixative and duration unreported (datasheet A08140-1); 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 A08140-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08140-1) |
| Primary antibody | Rabbit anti-DHX15, 2-5μg/ml (datasheet A08140-1) |
| Primary incubation | Overnight at 4 °C (datasheet A08140-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A08140-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DHX15-positive staining in purkinje cells of cerebellum (HPA tissue IHC: Medium). HPA tissue profile: Nuclear and nucleolar expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A08140-1). Sodium citrate retrieval was reported in another DHX15 IHC method (PMC6861564).
Section 2
What Is the Expected DHX15 Staining Pattern?
DHX15 is reported in the nucleus and nucleolus (UniProt O43143: subcellular location), with nuclear and nucleolar staining in several tissues (HPA: tissue IHC). Expect staining in selected glandular cells, pancreatic endocrine cells, ovarian follicle cells and cerebellar Purkinje cells (HPA: medium staining). Interpret this pattern cautiously: HPA rates the IHC antibody Approved, reports low consistency with RNA data and notes that external verification is pending (HPA: reliability). DHX15 has no transmembrane segment (UniProt O43143: topology).
| Nuclear or nucleolar signal in the listed positive cell populations. | This fits the reported tissue pattern; HPA records medium staining in those populations, without establishing identical intensity in every tissue (HPA: tissue IHC). |
| Predominantly cytoplasmic signal, with little nuclear staining. | Treat this as suspect: HPA observed and disregarded cytoplasmic staining, while UniProt places DHX15 in the nucleus and nucleolus (HPA: reliability; UniProt O43143: subcellular location). |
| Strong staining in cells listed as undetected, such as adipocytes or cardiomyocytes. | Check for cross-reactivity or endogenous detection activity before assigning DHX15 positivity; those particular cell populations were not detected by HPA IHC (HPA: tissue IHC; general IHC practice). |
| Diffuse chromogen across cells and surrounding tissue, without clear nuclear boundaries. | A compartment-free deposit does not match the reported nuclear and nucleolar pattern. Review background controls and the detection step before scoring cells (HPA: tissue IHC; general IHC practice). |
| No nuclear signal in a section containing a listed medium-staining population. | First check tissue identity, section quality and assay controls. A negative run in one section cannot alone overturn an HPA observation, especially given its stated validation caveat (HPA: tissue IHC and reliability; general IHC practice). |
💡Expected DHX15 appearanceCall a result positive when the appropriate cells show discernible nuclear or nucleolar staining, with medium staining reported for the listed positive populations; predominantly cytoplasmic or diffuse staining is suspect (HPA: tissue IHC and reliability).
| Choice of comparison tissue | Use a listed medium-staining population to judge whether the assay can show the expected compartment; HPA lists Purkinje cells and several glandular populations, but reports low staining in skin keratinocytes (HPA: tissue IHC). |
| Cell identity within a section | Score the named cells rather than an entire organ: HPA reports pancreatic endocrine cells as medium, for example, and reports liver cholangiocytes as not detected (HPA: tissue IHC). |
| Evidence strength | Approved is the HPA IHC rating for HPA047047; HPA also reports low agreement between antibody staining and RNA expression, with external verification pending (HPA: antibody validation and reliability). |
| DHX15 distribution and structure | UniProt calls tissue expression ubiquitous and reports no transmembrane segment. These facts support broad biological presence but do not predict equal visible IHC staining in every cell (UniProt O43143: tissue specificity and topology; HPA: tissue IHC). |
| IF/ICC question: where should signal appear? | HPA reports enhanced nuclear-speckle localization in ICC-IF; UniProt also lists nuclear and nucleolar locations. Compare each image with its assay's own controls rather than requiring identical IHC and IF detail (HPA: subcellular ICC-IF; UniProt O43143: subcellular location; general microscopy practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| The listed positive cells show no nuclear signal. | The section may lack the expected cells, or the staining run may have failed; absence alone does not identify the cause (HPA: tissue IHC; general IHC practice). | Confirm the named cell population and inspect a suitable positive control, then review the general IHC retrieval and detection workflow before interpreting the negative section (HPA: tissue IHC; general IHC practice). |
| Signal is mainly cytoplasmic. | HPA explicitly disregarded observed cytoplasmic staining, so it is weak evidence for DHX15 localization (HPA: reliability). | Check the negative control and antibody-dependent staining; score DHX15 only where a credible nuclear or nucleolar pattern is present (HPA: tissue IHC; general IHC practice). |
| An HPA-listed undetected cell type stains strongly. | Cross-reactivity or endogenous detection activity is possible; an HPA undetected call is also limited to its observed assay and sample (HPA: tissue IHC; general IHC practice). | Compare with a control lacking primary antibody and check whether staining follows cell boundaries or the reported nuclear compartment (HPA: tissue IHC; general IHC practice). |
| Chromogen appears broadly across the section. | Diffuse background can obscure compartment assignment and may reflect nonspecific binding or detection background (general IHC practice). | Inspect a control lacking primary antibody, blocking and detection conditions; score only resolved nuclear or nucleolar signal in the relevant cells (HPA: tissue IHC; general IHC practice). |
| Staining varies between named positive tissues. | HPA reports selected cells at medium intensity, other cells at low intensity or undetected, and low staining-to-RNA consistency (HPA: tissue IHC and reliability). | Record tissue, cell type, compartment and intensity separately; use the listed HPA observations as comparisons rather than assuming uniform staining (HPA: tissue IHC). |
| Nuclear puncta appear in IF/ICC but are less distinct in chromogenic IHC. | HPA describes enhanced nuclear speckles in ICC-IF and nuclear and nucleolar expression in tissue IHC; these are assay-specific observations (HPA: subcellular ICC-IF and tissue IHC). | Evaluate each assay against its reported pattern and controls; do not reject tissue IHC solely because it lacks the speckle detail reported for ICC-IF (HPA: subcellular ICC-IF and tissue IHC; general microscopy practice). |
Experiment evidence
IHC & IF images showing DHX15 staining
A08140-1 · IHCIHC analysis of DHX15/prp43 using anti-DHX15/prp43 antibody (A08140-1). DHX15/prp43 was detected in a paraffin-embedded section of human pro
A08140-1 · IHCIHC analysis of DHX15/prp43 using anti-DHX15/prp43 antibody (A08140-1). DHX15/prp43 was detected in a paraffin-embedded section of human tes
A08140-1 · IFIF analysis of DHX15/prp43 using anti-DHX15/prp43 antibody (A08140-1) and anti-Beta Tubulin antibody (M01857-3). DHX15/prp43 was detected in
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000109606-DHX15/tissue/cerebral+cortex#imid_13017191
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000109606-DHX15/tissue/colon#imid_13014173
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000109606-DHX15/tissue/liver#imid_13014182
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000109606-DHX15/tissue/kidney#imid_13014191
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA047047; green = DHX15, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA047047; green = DHX15, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DHX15 IHC & IF
🧪Run cerebellum first and score Purkinje cells, which show Medium DHX15 staining (HPA: cerebellum, Purkinje cells). Use adipose tissue as the negative comparator because adipocytes are Not detected (HPA: adipose tissue, adipocytes); on the positive slide, cells scored as unstained should show counterstain without nuclear DAB signal, but should not be assumed to be verified DHX15-negative cells (standard IHC practice).
Positive control tissue: Cerebellum (Purkinje cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show DHX15 in A-431, HEK293, U2OS, U2OS, scrambled (10x), U2OS, siRNA 1 (10x), NIH 3T3, with annotated localisation: Nuclear speckles (enhanced) (HPA subcellular).
Technical controls: Include no-primary (secondary-only), concentration-matched rabbit IgG isotype control matched to the primary’s clonality, and, if available, identically processed DHX15-knockout material as a biological negative (caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase before HRP/DAB detection in cerebellum; for IF, check neuronal autofluorescence (caption: HRP/DAB; standard IHC/IF practice).
⚠️Feasibility: No supplied source reports a DHX15-specific fixation window, fixation effect, or retrieval dependency; the fixative is unreported in the selected A08140-1 paraffin-section caption (A08140-1 tissue-IHC caption). That caption reports heat-mediated retrieval in EDTA, pH 8.0, which provides a starting condition for IHC optimization (A08140-1 tissue-IHC caption). IF/ICC can assess the reported nuclear-speckle pattern, but the supplied evidence does not establish that IF or frozen sections are easier; cerebellar neuronal autofluorescence may complicate IF interpretation (HPA subcellular: nuclear speckles; standard IF practice).
HPA tissue IHC evidence for DHX15
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. Pending external verification. Cytoplasmic staining observed and disregarded.). 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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | Medium | Protein (IHC) | HPA → |
| Ovary | Follicle cells | Medium | Protein (IHC) | HPA → |
| Pancreas | Pancreatic endocrine 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 → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced DHX15 IHC Tips
Troubleshoot DHX15 staining in paraffin-section chromogenic IHC using the catalog antibody’s tissue image, reported localisation, and matched controls.
What retrieval should I try first when DHX15 staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A08140-1). The catalog image used this retrieval before 2 μg/ml primary antibody overnight at 4°C, so retain those conditions while assessing retrieval (datasheet A08140-1). Compare staining in the same cell type across adjacent sections, and examine nuclear detail and section integrity before increasing heat exposure (standard IHC practice; UniProt O43143 localisation). If staining remains weak, test a different retrieval buffer as a controlled fallback, recording heat time and temperature for each condition (standard IHC practice).
Could fixation explain weak DHX15 staining in my paraffin sections?
DHX15-specific sensitivity to fixation is unknown because the selected tissue image reports a paraffin-embedded section but does not state its fixative (datasheet A08140-1). Record the specimen’s fixative and fixation duration, then compare similarly processed sections while holding retrieval at EDTA pH 8.0 constant (standard IHC practice; datasheet A08140-1). If fixation histories differ, assess staining and tissue morphology together before attributing an intensity difference to DHX15 abundance (standard IHC practice). Do not infer a fixation effect from DHX15’s nuclear localisation, lack of a transmembrane segment, or reported phosphorylation and acetylation (UniProt O43143).
Where should convincing DHX15 staining appear in tissue?
Prioritise nuclear staining, including nucleoli, when reviewing chromogenic tissue sections (UniProt O43143 localisation; HPA tissue IHC). HPA reports nuclear and nucleolar expression in several tissues, while its cell imaging places DHX15 in nuclear speckles (HPA tissue IHC; HPA subcellular). Inspect high-power fields to determine whether DAB follows nuclei rather than section folds, tissue edges, or diffuse extracellular material (standard IHC practice). Treat predominantly cytoplasmic staining cautiously: HPA explicitly disregarded its observed cytoplasmic staining, and its tissue antibody assessment remains pending external verification (HPA tissue IHC).
How should I investigate an unexpected DHX15 staining pattern?
First check the antibody’s documented immunogen or epitope against the target sequence; no epitope position is supplied here (standard IHC practice; datasheet A08140-1). The supplied DHX15 record lists 0 isoforms and one chain spanning residues 1–795, so an alternative isoform is not supported as the explanation (UniProt O43143). Its ATP-binding and helicase C-terminal domains span residues 147–313 and 338–518, respectively, but those domain positions do not identify the antibody’s binding site (UniProt O43143). Compare retrieval conditions and nuclear staining in adjacent sections before assigning a pattern change to epitope masking or a modification (standard IHC practice; UniProt O43143 localisation).
How can I assess DHX15 by multiplex immunofluorescence?
Validate the antibody and staining conditions for IF separately from its paraffin-section chromogenic image (datasheet A08140-1; standard IF practice). Pair DHX15 with a validated marker for the expected cell type, such as follicle cells in ovary, and use a nuclear counterstain to assess compartment assignment (HPA tissue IHC; standard IF practice). Choose a far-red fluorophore when tissue autofluorescence compromises shorter wavelengths, and include single-label controls for channel separation (standard IF practice). Because DHX15 is nuclear and has no transmembrane segment, optimise permeabilisation for nuclear antibody access while checking that nuclear morphology remains intact (UniProt O43143 topology and localisation; standard IF practice).
What should I check when DAB obscures DHX15-positive nuclei?
Run a section without primary antibody to assess secondary reagent, chromogen, and tissue-derived background before changing the primary concentration (standard IHC practice). Include an endogenous peroxidase block in the chromogenic workflow, and examine whether brown signal persists outside nuclei after that control (standard IHC practice; UniProt O43143 localisation). The selected image used 10% goat serum, 2 μg/ml primary antibody, a peroxidase-conjugated secondary, and DAB; these are documented image conditions rather than background guarantees (datasheet A08140-1). If diffuse staining remains, compare antibody concentration and wash conditions on adjacent sections while checking that nuclear signal is retained (standard IHC practice).
How should I score DHX15 across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score nuclear staining because the reported tissue pattern is nuclear and nucleolar (HPA tissue IHC; UniProt O43143 localisation). Record the percentage of positive nuclei and intensity categories, then calculate an H-score using the percentage in each category multiplied by its intensity score (standard IHC practice). Normalise counts to the number of eligible cells in the scored compartment, or report positive-cell density per mm² when sampling area is the comparison unit (standard IHC practice). Keep retrieval, DAB development, imaging, and scoring thresholds consistent across samples, and record excluded damaged or necrotic regions (standard IHC practice).
How do I distinguish true DHX15 staining from artefact?
A credible result follows nuclear or nucleolar boundaries in intact cells, consistent with the reported DHX15 tissue distribution (HPA tissue IHC; UniProt O43143 localisation). Compare the relevant cell type with a documented reference, such as medium staining in cerebellar Purkinje cells, without assuming that every cell in that tissue is positive (HPA tissue IHC). Reconsider diffuse cytoplasmic colour, strong section-edge signal, necrotic regions, or staining retained in a no-primary control; HPA disregarded cytoplasmic staining in its assessment (HPA tissue IHC; standard IHC practice). Check endogenous peroxidase blocking and section morphology before interpreting DAB intensity as biological variation (standard IHC practice).
Boster reagents
Best DHX15 / ATP-dependent RNA helicase DHX15 IHC Antibodies
A08140-1 has paraffin-section IHC images from human tumors and an IF/ICC image from U2OS cells (catalog: IHC and IF image captions). Listed reactivity includes human, mouse and rat (catalog: reactivity).
Real IHC data
Anti-DHX15/prp43 Antibody ®
Cat # A08140-1
A08140-1 will render with its human prostate cancer paraffin-section IHC figure (catalog: IHC image caption). Its additional IHC captions cover human testicular germ cell tumor, colorectal adenocarcinoma and liver cancer; its IF caption shows U2OS cells (catalog: IHC and IF image captions).
Which to pick: Choose A08140-1 for paraffin-section tissue IHC: its figure documents EDTA pH 8.0 retrieval, 2 μg/ml primary antibody and DAB detection; the fixative is unreported (catalog: IHC image caption). Choose the same SKU for IF/ICC based on its U2OS cell image and 5 μg/ml IF concentration (catalog: IF image caption). Human, mouse and rat reactivity is listed, but the supplied IHC and IF images show human samples only; the antibody is rabbit-hosted and its clone is unreported (catalog: reactivity, host and clone; IHC and IF image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O43143 (DHX15_HUMAN, ATP-dependent RNA helicase DHX15).
- Human Protein Atlas. DHX15 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. DHX15 subcellular location (ICC-IF): Localized to the nuclear speckles..
- Human Protein Atlas. DHX15 antibody validation summary (1 antibodies).
- Long Non-coding RNA JHDM1D-AS1 Interacts with DHX15 Protein to Enhance Non-Small-Cell Lung Cancer Growth and Metastasis. Molecular therapy. Nucleic acids 2019 — PMC6861564.
- DHX15 Inhibits Autophagy and the Proliferation of Hepatoma Cells. Frontiers in medicine 2020 — PMC7904900.
- Fusobacterium nucleatum promotes tumor progression in KRAS p.G12D-mutant colorectal cancer by binding to DHX15. Nature communications 2024 — PMC10894276.
- RNA helicase DHX15 decreases cell apoptosis by NF-κB signaling pathway in Burkitt lymphoma. Cancer cell international 2022 — PMC8862312.
- PubMed PMID:9388478 — UniProt-cited evidence.
- PubMed PMID:12458796 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DHX15 (ATP-dependent RNA helicase DHX15). Expected localisation Nuclear and nucleolar staining in several tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A08140-1). Validated IHC antibody: A08140-1 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human prostate cancer tissue; fixative not specified (datasheet A08140-1); 2. Fixation: Image fixative and duration unreported (datasheet A08140-1); 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 A08140-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08140-1); 8. Primary antibody: Rabbit anti-DHX15, 2-5μg/ml (datasheet A08140-1); 9. Primary incubation: Overnight at 4 °C (datasheet A08140-1). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Cytoplasmic staining was observed and disregarded (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one full-length chain (UniProt); Regulation: No expression regulator is specified (UniProt). Evidence: UniProt O43143, HPA tissue IHC · Approved, HPA ICC-IF · nuclear speckles., Datasheet A08140-1, PMC6861564, PMC10894276. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DHX15/immunohistochemistry-ihc-guide.