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DHX15 / ATP-dependent RNA helicase DHX15 · IHC design guide
Design Immunohistochemistry for DHX15
Plan DHX15 staining in paraffin sections around the nuclear and nucleolar pattern observed in several tissues (HPA tissue IHC). Start with the catalog antibody at 2–5 μg/ml (datasheet A08140-1) and assess nuclear staining with appropriate controls (standard IHC practice).
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/nucleolar signal in rectal glandular cells (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 sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A08140-1) | |
| Caveat | Cytoplasmic staining was observed and disregarded (HPA tissue IHC) | |
| Regulation | Ubiquitous expression; no inducer stated (UniProt) | |
| Isoform / epitope | No annotated isoforms or cleavage; no membrane epitope split (UniProt) |
Section 1
Recommended DHX15 IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 retrieval (datasheet A08140-1). These 3 published DHX15 IHC methods provide additional examples (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 noteTry EDTA pH 8.0 heat retrieval first with the catalog antibody (datasheet A08140-1). Sodium citrate retrieval was used with a different DHX15 antibody (PMC6861564).
Section 2
What Is the Expected DHX15 Staining Pattern?
DHX15 is a nuclear and nucleolar protein with no transmembrane segment (UniProt O43143 localisation and topology). In paraffin sections, expect nuclear or nucleolar staining in several cell types, including Purkinje cells and glandular cells in stomach, rectum and gallbladder (HPA: tissue IHC). HPA rates the tissue pattern Approved but reports low consistency with RNA expression, pending external verification, and disregards observed cytoplasmic staining (HPA: IHC reliability).
| Distinct nuclear or nucleolar staining in Purkinje cells or stomach, rectum or gallbladder glandular cells. | This fits the reported medium staining in those cells and the expected compartment (HPA: tissue IHC; UniProt O43143 localisation). Assess the positive cells individually; the HPA profile describes expression in several tissues, while its RNA comparison has low consistency (HPA: IHC reliability). |
| Predominantly cytoplasmic staining, with little convincing nuclear signal. | Do not score cytoplasm alone as a confirmed DHX15 positive result: HPA observed cytoplasmic staining and disregarded it (HPA: IHC reliability). Review staining specificity and detection background before interpreting the section, because the reported protein location is nuclear and nucleolar (UniProt O43143 localisation). |
| Staining appears mainly in an HPA-listed, undetected cell type, such as adipocytes or cardiomyocytes. | Treat the mismatch as a reason to investigate cross-reactivity or endogenous detection activity (HPA: adipocytes and cardiomyocytes not detected; general IHC practice). It is not proof of either: DHX15 is described as ubiquitous, and HPA flags limited agreement between staining and RNA data (UniProt O43143 tissue specificity; HPA: IHC reliability). |
| Broad colour deposition obscures nuclei, tissue edges or empty areas. | A compartment cannot be scored reliably when diffuse background masks cell boundaries (general IHC practice). Check a no-primary control and the detection and blocking steps; background by itself does not establish DHX15 expression (general IHC practice). |
| No nuclear signal in a section expected to be positive, such as stomach glandular cells. | HPA reports medium staining in stomach glandular cells, so a blank result calls for a run-level check (HPA: stomach glandular cells). Confirm that tissue and detection controls worked, then review the antibody and antigen-retrieval workflow (general IHC practice). |
💡Expected DHX15 appearanceCall a section positive when identifiable cells show nuclear or nucleolar staining, with medium signal expected in the listed glandular cells and Purkinje cells; isolated cytoplasmic colour is a suspect result (HPA: tissue IHC and disregarded cytoplasmic staining; UniProt O43143 localisation).
| Tissue and cell selection | Stomach, rectum and gallbladder glandular cells and cerebellar Purkinje cells offer reported medium IHC examples (HPA: tissue IHC). An HPA 'not detected' cell type is a comparison, not proof that DHX15 is absent, given the ubiquitous tissue-specificity annotation (UniProt O43143 tissue specificity). |
| Compartment and topology | Score nuclei and nucleoli because those are the reported locations (UniProt O43143 localisation; HPA: tissue IHC). DHX15 has no transmembrane segment, so a membrane-only pattern lacks support from the supplied record (UniProt O43143 topology). |
| Antibody evidence | HPA047047 is Approved for tissue IHC and Enhanced for ICC-IF (HPA: antibody validation). The tissue IHC profile still carries a low RNA-consistency warning and pending external verification; the ICC-IF rating does not remove that IHC caveat (HPA: IHC reliability and antibody validation). |
| IF/ICC Q: where should DHX15 appear? | A: HPA reports enhanced localisation to nuclear speckles in ICC-IF (HPA: subcellular ICC-IF). Use that as an IF/ICC interpretation reference; assess paraffin-section IHC against its nuclear and nucleolar tissue profile (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected-positive glandular cells have no signal. | A failed stain or detection run is possible; HPA reports medium staining in stomach, rectum and gallbladder glandular cells (HPA: tissue IHC; general IHC practice). | Check section integrity and run controls, then review the validated antibody and antigen-retrieval steps used for the assay (general IHC practice). |
| Cytoplasmic colour dominates the section. | HPA observed cytoplasmic staining but disregarded it when describing the DHX15 tissue pattern (HPA: IHC reliability). | Record nuclear and cytoplasmic signals separately; compare with a no-primary control before assigning a positive result (general IHC practice). |
| Undetected reference cells stain strongly. | Cross-reactivity or endogenous detection activity is possible; HPA lists adipocytes and cardiomyocytes as not detected (HPA: tissue IHC; general IHC practice). | Compare matched no-primary and detection controls, and confirm the signal's compartment before interpreting the mismatch (general IHC practice). |
| Diffuse deposit makes nuclei hard to distinguish. | Background from the staining or detection workflow can obscure localisation (general IHC practice). | Inspect the no-primary control and review blocking, washing and chromogen development for the run (general IHC practice). |
| A low-staining tissue looks negative. | HPA reports low staining in hippocampal neuronal cells and duodenal glandular cells, so signal may be difficult to distinguish from background (HPA: tissue IHC; general IHC practice). | Interpret alongside a reported medium tissue control and avoid calling absence from that low-staining field alone (HPA: tissue IHC; general IHC practice). |
| IF/ICC speckles and paraffin IHC look different. | HPA describes nuclear speckles in ICC-IF and nuclear and nucleolar expression in tissue IHC (HPA: subcellular ICC-IF and tissue IHC). | Judge each preparation against its own reported pattern; do not require resolved speckles to score a tissue-IHC nucleus (HPA: subcellular ICC-IF and tissue IHC). |
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 look for staining in Purkinje cells (HPA: Medium in cerebellar Purkinje cells). Run adipose tissue as the negative comparator, focusing on adipocytes (HPA: Not detected in adipocytes); on the cerebellum slide, use neighboring cells without visible signal to assess background, without assuming they are DHX15-negative.
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 a no-primary, secondary-only control, a matched rabbit IgG isotype control (caption: rabbit primary antibody), and DHX15-knockout material as a biological specificity control (standard IHC practice). For chromogenic detection, block endogenous peroxidase and inspect the cerebellum slide for background staining (caption: peroxidase secondary and DAB; standard IHC practice).
⚠️Feasibility: The selected paraffin-section example uses heat retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required across specimens (caption: EDTA retrieval). Its fixative is unreported, and no target-specific fixation window or fixation effect is supplied (caption: fixative not stated). Neither frozen sections nor IF can be judged easier from these data; for cerebellar IF, check whether neuronal autofluorescence obscures the signal (HPA: cerebellar Purkinje-cell staining; 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 by checking retrieval, compartment, controls, and scoring against the expected nuclear and nucleolar pattern (UniProt O43143; HPA tissue IHC).
How should I adjust retrieval when DHX15 staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA, pH 8.0, for paraffin sections (datasheet A08140-1). The selected tissue image used this retrieval before an overnight incubation at 4°C with 2 μg/ml primary antibody (caption A08140-1). If nuclear staining remains weak, compare retrieval times such as 10 and 20 minutes on adjacent sections while keeping antibody concentration and detection constant; these times are optimisation trials, not reported product conditions. Check tissue integrity and nuclear contrast after each trial, then select the shortest treatment that gives reproducible nuclear staining with little background (UniProt O43143 localisation; HPA tissue IHC).
Could fixation explain absent DHX15 staining in my paraffin sections?
Target-specific fixation sensitivity is unknown: the selected DHX15 image identifies paraffin-embedded tissue but does not state its fixative (caption A08140-1). Record the fixative and time for each specimen, and compare similarly processed sections before attributing a negative result to DHX15 abundance. If you control fixation, compare a short and a longer interval under the same routine fixative, then apply the same EDTA, pH 8.0 retrieval to both (datasheet A08140-1). Inspect morphology alongside staining, and include a section showing the expected nuclear or nucleolar signal as a process control (UniProt O43143 localisation; HPA tissue IHC).
What staining pattern should I expect, and how should I assess diffuse cytoplasmic signal?
Evaluate DHX15 primarily in nuclei and nucleoli of intact cells (UniProt O43143 localisation; HPA tissue IHC). Nuclear speckles are also reported by subcellular imaging, although their fine pattern may be difficult to resolve with chromogenic tissue staining (HPA subcellular). The tissue atlas specifically disregarded observed cytoplasmic staining, so score diffuse cytoplasmic DAB separately from nuclear signal (HPA tissue IHC). On adjacent sections, compare a no-primary control and the 2 μg/ml primary-antibody condition under identical development times; inspect whether the suspect signal follows tissue damage or persists without primary antibody (caption A08140-1; standard IHC controls).
How can I troubleshoot inconsistent staining if the antibody epitope is unknown?
The supplied record lists 0 isoforms and a 1–795 chain, but it does not map the catalog antibody's epitope (UniProt O43143; caption A08140-1). DHX15 has annotated phosphoserine at residue 15 and acetyllysine at 488; neither annotation establishes an effect on this antibody's binding (UniProt O43143). Compare adjacent sections with identical EDTA, pH 8.0 retrieval and detection settings before assigning inconsistent staining to epitope access (datasheet A08140-1). If an independently validated antibody with a disclosed epitope is available, compare nuclear patterns and document each antibody's controls before interpreting discordance as biology (UniProt O43143 localisation; standard IHC practice).
How should I plan multiplex IF to check the DHX15 pattern?
For a separate IF experiment, pair DHX15 with a validated marker identifying the cell population of interest; ovarian follicle cells are one population with medium tissue staining (HPA tissue IHC). Choose a far-red channel for DHX15 when tissue autofluorescence is prominent, and image unstained and single-stain controls to set exposure and assess bleed-through (standard IF practice). DHX15 lacks a transmembrane segment and is reported in nuclei and nucleoli, so use controlled permeabilisation to reach its intracellular epitope, for example a trial of 0.1% Triton X-100 for 5 minutes (UniProt O43143 topology and localisation; standard IF practice). Check for nuclear speckles against the subcellular reference, while validating the chosen IF antibody independently of the paraffin IHC caption (HPA subcellular; caption A08140-1).
How can I separate nonspecific DAB staining from DHX15 signal?
Run a no-primary section through the same secondary antibody, peroxidase detection, and DAB development to reveal detection-related background (standard IHC practice). The selected image used 10% goat serum blocking, a peroxidase-conjugated secondary incubated for 30 minutes at 37°C, and DAB development (caption A08140-1). Apply an endogenous-peroxidase block as a general chromogenic IHC step, and compare staining before and after a shorter DAB development if diffuse colour persists (standard IHC practice). Keep the assessment anchored to nuclei and nucleoli, because cytoplasmic staining was disregarded in the tissue atlas (UniProt O43143 localisation; HPA tissue IHC).
What is a defensible way to quantify DHX15 chromogenic staining? ⚠ ANSWER MARKED FOR VERIFICATION
Segment viable cells by tissue region and score nuclear DHX15 separately from cytoplasmic colour, using the expected nuclear and nucleolar pattern as the compartment rule (UniProt O43143 localisation; HPA tissue IHC). Report the percentage of positive nuclei and an H-score from 0–300, with intensity categories defined before reviewing groups (standard IHC scoring practice). Normalise positive counts to the number of evaluable nuclei in each region, and report sampling area in mm² if using positive-cell density (standard IHC quantification practice). Keep retrieval, antibody concentration, DAB development, and image thresholds constant across specimens; document exclusions for damaged or necrotic areas (datasheet A08140-1; standard IHC practice).
When does a DHX15-positive section support a biological conclusion?
Give the most weight to reproducible nuclear or nucleolar staining in morphologically intact cells, consistent with the reported compartment (UniProt O43143 localisation; HPA tissue IHC). Treat isolated cytoplasmic colour cautiously: the tissue atlas observed and disregarded cytoplasmic staining, and its tissue IHC reliability is Approved with low consistency against RNA data and pending external verification (HPA tissue IHC). Inspect section edges, necrotic regions, and the no-primary control for artefact or endogenous enzyme signal before calling a region positive (standard IHC practice). Compare like cell populations across specimens; a negative cell type does not establish global absence, because the atlas reports both stained and undetected populations (HPA tissue IHC).
Boster reagents
Best DHX15 / ATP-dependent RNA helicase DHX15 IHC Antibodies
A08140-1 has IHC images from human paraffin sections and an IF/ICC image from U2OS cells (catalog image captions); its listed reactivity is human, mouse and rat (catalog).
Real IHC data
Anti-DHX15/prp43 Antibody ®
Cat # A08140-1
A08140-1 has IHC images from paraffin sections of human prostate cancer, testicular germ cell tumor, colorectal adenocarcinoma and liver cancer tissue (catalog IHC captions). Its IF/ICC image shows U2OS cells, and IF and ICC are listed applications (catalog IF caption; catalog applications).
Which to pick: Choose A08140-1 for human tissue IHC on paraffin sections because its IHC captions document staining with EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (catalog IHC captions). Choose A08140-1 for IF/ICC because its IF caption documents U2OS cell staining at 5 μg/ml (catalog IF caption). The catalog lists human, mouse and rat reactivity, but the supplied IHC and IF images show human samples only; the tissue fixative and antibody clonality are unreported (catalog reactivity; catalog image captions; catalog clone field).
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 sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A08140-1); Caveat: Cytoplasmic staining was observed and disregarded (HPA tissue IHC); Isoform / epitope: No annotated isoforms or cleavage; no membrane epitope split (UniProt); Regulation: Ubiquitous expression; no inducer stated (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.