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DPYSL5 / Dihydropyrimidinase-related protein 5 · IHC design guide
Design Immunohistochemistry for DPYSL5
Plan DPYSL5 paraffin IHC around cytoplasmic staining in CNS and testis (HPA tissue IHC). Use caudate neurons as a positive reference (HPA tissue IHC), and start the IHC-P antibody M08644-1 at 1:25 (datasheet M08644-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DPYSL5 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 staining in CNS and testis (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic in caudate neurons and testis spermatids (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet M08644-1) | |
| Positive control | Caudate+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Formaldehyde-fixed paraffin sections were used (selected-SKU IHC image M08644-1) | |
| Caveat | Cerebellar granular cells were undetected (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | No annotated isoforms or processing; epitope impact unknown (UniProt) |
Section 1
Recommended DPYSL5 IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet: M08644-1); the published DPYSL5 IHC methods below provide two additional tissue-staining examples (PMC10796342; PMC5319751).
| Sample | Formaldehyde-fixed, paraffin-embedded tissue sections (datasheet M08644-1; sample unspecified) |
| Fixation | Image formalin-fixed; duration unreported (datasheet M08644-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6 (datasheet M08644-1); 20 min, 95–100 °C (standard) |
| 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-DPYSL5, 1:25 (datasheet M08644-1) |
| Primary incubation | 1 hours at 37°C (datasheet M08644-1) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DPYSL5-positive staining in neuronal cells of caudate (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in CNS and testis. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet: M08644-1); the brain-tissue protocol also used citrate pH 6.0 (PMC5319751).
Section 2
What Is the Expected DPYSL5 Staining Pattern?
DPYSL5 is annotated as cytoplasmic and has no transmembrane segment (UniProt Q9BPU6). In tissue IHC, expect cytoplasmic staining in caudate neuronal cells, cortical and hippocampal glial cells, and late spermatids; enterocytes also show staining (HPA tissue IHC). HPA rates the tissue pattern Enhanced, citing agreement with RNA data and GTEx support in the CNS (HPA tissue IHC).
| Strong cytoplasmic stain in caudate neuronal cells. | This matches the reported High neuronal staining and cytoplasmic tissue profile (HPA tissue IHC). Judge the pattern in identifiable cells, since a dark area without clear cellular boundaries gives less confidence that the chromogen marks DPYSL5. |
| Moderate cytoplasmic stain in cerebral cortex or hippocampal glial cells, testicular late spermatids, or small-intestinal enterocytes. | Each listed cell population is reported at Medium intensity (HPA tissue IHC). A weaker result than caudate neurons can therefore fit the reference pattern; compare corresponding cell types rather than expecting every positive tissue to stain equally. |
| Predominantly nuclear stain in a paraffin tissue section, with little cytoplasmic signal. | This differs from the reported cytoplasmic tissue IHC profile (HPA tissue IHC; UniProt Q9BPU6). Investigate the tissue pattern and controls before assigning it to DPYSL5. Nuclear staining alone is not proof of artefact: HPA separately approves nucleoplasmic localization in ICC-IF (HPA subcellular). |
| Stain concentrated in an HPA-listed negative cell population, such as adipocytes in adipose tissue or cerebellar granular-layer cells. | Those specific populations are reported Not detected (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity, and review morphology and controls. The HPA result applies to the named cells; it does not declare every cell in the organ negative. |
| Widespread haze across tissue and surrounding section, or no stain in caudate neuronal cells. | A haze obscures the expected cellular pattern and may reflect nonspecific staining or detection background (general IHC practice). No caudate neuronal signal conflicts with the reported High level (HPA tissue IHC); first establish that tissue, staining run, and controls performed adequately. |
💡Expected DPYSL5 appearanceCall the IHC result convincing when cytoplasmic stain follows the reported cells—strong in caudate neurons and moderate in the listed glia, spermatids, or enterocytes (HPA tissue IHC); uniform haze or dominant stain in listed negative cells warrants investigation.
| Tissue and cell selection | Caudate neurons offer a High reference; cortical and hippocampal glia, late spermatids, and enterocytes offer Medium references (HPA tissue IHC). Use the named cell population when judging a section; whole-organ labels are too broad for these comparisons. |
| IHC versus ICC-IF localization | Tissue IHC is summarized as cytoplasmic, whereas ICC-IF lists nucleoplasm as the approved main location and cytosol as an approved additional location (HPA tissue IHC; HPA subcellular). An IF image therefore needs its own interpretation; this IHC guide supplies no ICC-IF protocol. |
| Antibody evidence | HPA034544 is rated Enhanced for IHC, while HPA072387 is listed as Approved for ICC with no IHC rating in the supplied record (HPA antibodies). Match the validation to the application; the ICC rating alone does not establish an IHC staining pattern. |
| Topology and processing | DPYSL5 has no transmembrane segment or annotated signal peptide, propeptide, or cleavage into multiple chains; the recorded chain spans residues 1–564 (UniProt Q9BPU6). A cell-surface outline alone would be unexpected from this record and should prompt a control review. |
| Epitope and preparation limits | Five modified residues are annotated, but no antibody epitope, retrieval condition, or target-specific fixation effect is supplied (UniProt Q9BPU6; HPA antibodies). Do not attribute a weak stain to a particular modification or fixation effect without separate evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| Caudate neuronal cells show no convincing stain. | The result conflicts with their reported High level (HPA tissue IHC); the record does not identify a DPYSL5-specific preparation failure. | Check tissue identity and run controls, then review the IHC-validated antibody's specified paraffin-section procedure and the detection steps (general IHC practice). |
| A brown haze covers positive and negative regions. | Possible nonspecific binding, excess reagent, or detection background (general chromogenic IHC practice); haze cannot be scored as the cytoplasmic pattern (HPA tissue IHC). | Review blocking, washing, antibody concentration, and no-primary control; assess stain only where cell boundaries and compartment remain clear (general IHC practice). |
| Adipose adipocytes or cerebellar granular-layer cells stain prominently. | These named cells are reported Not detected (HPA tissue IHC). Cross-reactivity or endogenous detection activity are possibilities, not established causes. | Confirm cell identity; compare a no-primary control and, where relevant, an endogenous-enzyme block before interpreting the stain as DPYSL5 (general chromogenic IHC practice). |
| Nuclear stain dominates a tissue IHC section. | That differs from the cytoplasmic tissue profile, although nucleoplasm is an approved ICC-IF location (HPA tissue IHC; HPA subcellular). | Check whether cytoplasmic signal is present in known-positive cells, inspect controls, and report the IHC–ICC-IF difference without treating the IF result as an IHC validation. |
| A testis or small-intestine section is scored entirely negative or entirely positive. | HPA reports Medium staining in late spermatids or enterocytes specifically, not a uniform organ-wide signal (HPA tissue IHC). | Locate the named cell population before scoring; compare its cytoplasm with neighboring cells and the run controls (HPA tissue IHC; general IHC practice). |
| Only a sharp cell-edge outline appears, without cytoplasmic stain. | The outline does not match the cytoplasmic tissue profile or the lack of a transmembrane segment (HPA tissue IHC; UniProt Q9BPU6). | Review morphology, no-primary control, and detection background; avoid assigning the edge signal to DPYSL5 unless independent evidence supports it (general IHC practice). |
Experiment evidence
IHC & IF images showing DPYSL5 staining
M08644-1 · IHCM08644-1 staining DPYSL5 in human tonsil sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections). Tissu
M08644 · IFImmunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (human neuroblastoma cell line) cells label
M08644-2 · IFImmunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (human neuroblastoma cell line) cells label
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000157851-DPYSL5/tissue/cerebral+cortex#imid_19963293
HPA · CaudateHuman Protein Atlas IHC, Caudate. Source: https://www.proteinatlas.org/ENSG00000157851-DPYSL5/tissue/basal+ganglia#imid_19963295
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000157851-DPYSL5/tissue/colon#imid_19963227
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000157851-DPYSL5/tissue/liver#imid_19963228
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA072387; green = DPYSL5, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA072387; green = DPYSL5, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DPYSL5 IHC & IF
🧪Run caudate first and expect neuronal cells to stain (HPA: caudate neuronal cells High); use adipose tissue as the negative tissue and expect adipocytes to lack staining (HPA: adipose adipocytes Not detected). On the caudate slide, neighboring cells without signal can serve as an internal background reference, but do not assume a particular non-neuronal cell type is negative from the supplied HPA row (HPA: caudate neuronal cells High).
Positive control tissue: Caudate (Neuronal cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show DPYSL5 in HEK293, SH-SY5Y, U2OS, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species and clonality; use DPYSL5 knockout material or a validated peptide block as a biological specificity control (standard IHC practice). For chromogenic caudate IHC, block endogenous peroxidase and check endogenous biotin when using a biotinylated secondary; assess pigment against the no-primary slide (selected IHC caption: biotinylated secondary; standard IHC practice).
⚠️Feasibility: A DPYSL5-specific fixation window or fixation effect is unreported, and the selected M08644-1 tissue-IHC record lists the fixative as unreported (selected-SKU fixative field: not stated). Heat retrieval in citrate buffer at pH 6 is described, but a requirement for retrieval is untested; the caption’s primary dilution is 1:25 (selected IHC caption). The supplied evidence does not establish whether frozen sections or IF/ICC are easier; caudate pigment may complicate chromogenic scoring, while HPA ICC-IF images show nucleoplasm and cytosol localization in HEK293, SH-SY5Y and U2OS (standard IHC practice; HPA: subcellular summary and ICC-IF cell lines).
HPA tissue IHC evidence for DPYSL5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. GTEX supports antibody staining in CNS.). 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 |
|---|---|---|---|---|
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | Medium | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Medium | Protein (IHC) | HPA → |
| Small intestine | Enterocytes | Medium | Protein (IHC) | HPA → |
| Testis | Elongated or late spermatids | 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 | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced DPYSL5 IHC Tips
Troubleshoot DPYSL5 chromogenic IHC by checking retrieval, tissue controls, cell identity and staining compartment; use the IF guidance for complementary localisation checks.
How should I optimise retrieval when DPYSL5 staining is weak in paraffin sections?
Use heat-mediated citrate buffer at pH 6 for DPYSL5 antigen retrieval in paraffin sections (datasheet M08644-1). Keep buffer, section thickness and detection conditions constant while comparing 10 and 20 minutes of heating, followed by a 20-minute cooldown (standard IHC optimisation practice). Include caudate neuronal cells as a tissue positive control when available (HPA tissue IHC: High in caudate neuronal cells). Assess whether stronger signal also raises diffuse background or damages section morphology before selecting a heating time (standard IHC practice). Record the chosen time and heating device so subsequent slides receive the same retrieval treatment (standard IHC practice).
Could fixation explain variable DPYSL5 staining between paraffin blocks?
Target-specific DPYSL5 sensitivity to fixative choice or fixation duration is unknown because comparative fixation evidence is not supplied (evidence payload). For a new tissue series, record fixative, time to fixation and fixation duration, then process matched samples together where feasible (standard IHC practice). A pilot comparing 6 and 24 hours in 10% neutral buffered formalin can reveal whether the chosen preparation changes signal or morphology in that series (standard IHC optimisation practice). Apply the same citrate pH 6 retrieval and detection settings to both preparations (datasheet M08644-1; standard IHC practice). Interpret any difference as a result of that pilot, not an established DPYSL5 fixation effect (evidence payload).
Should DPYSL5 IHC signal appear in cytoplasm or nuclei?
Expect a substantial cytoplasmic component in tissue IHC, particularly in CNS and testis, while examining the labelled cell type (HPA tissue IHC: cytoplasmic expression in CNS and testis). UniProt also annotates DPYSL5 as cytoplasmic and without a transmembrane segment (UniProt Q9BPU6 localisation and topology). Nuclear signal warrants review, but it cannot be dismissed solely by compartment: HPA subcellular imaging reports approved nucleoplasmic and additional cytosolic localisation (HPA subcellular). Compare nuclear and cytoplasmic staining separately against the same-run controls, including secondary-only slides (standard IHC practice). At 40×, confirm that apparent nuclear colour lies within intact cells rather than overlying pigment or precipitate (standard IHC practice).
How can epitope uncertainty affect interpretation of DPYSL5 IHC?
The supplied record annotates 0 isoforms and one DPYSL5 chain spanning residues 1–564; it does not identify this antibody’s epitope (UniProt Q9BPU6 record; evidence payload). Therefore, do not assign a staining pattern to a particular splice form or claim that the antibody distinguishes modified from unmodified DPYSL5 (evidence payload). The record includes phosphorylation at residues 509, 514, 532 and 538, plus methylation at 559 (UniProt Q9BPU6 modified residues). If staining changes across preparations, test matched blocks under identical retrieval and detection conditions before attributing it to epitope masking (standard IHC practice). Seek documented epitope information or an independently validated antibody before making site-specific claims (standard antibody validation practice).
How can IF help check cell identity and localisation seen by DPYSL5 IHC?
Use IF as a complementary check of the cell type and compartment assigned in chromogenic DPYSL5 IHC (standard IHC/IF practice). In neuronal regions, multiplex DPYSL5 with a neuronal marker such as MAP2 and score cells with both signals; caudate neuronal cells show high tissue staining (HPA tissue IHC: caudate neuronal cells; standard IF practice). Choose red or far-red fluorophores and inspect unstained tissue first when green autofluorescence obscures signal (standard IF practice). Apply a mild permeabilisation pilot, such as 0.1% Triton X-100, for intracellular access; DPYSL5 has no transmembrane segment (standard IF practice; UniProt Q9BPU6 topology). Check nuclear and cytosolic channels separately because both locations appear in subcellular imaging (HPA subcellular).
What should I check if DPYSL5 IHC has widespread brown background?
First compare the stained slide with a no-primary control to separate primary-dependent signal from detection background (standard IHC practice). The selected catalog image used a biotinylated secondary, making endogenous biotin a relevant control issue for that detection workflow (caption M08644-1; standard IHC practice). If using peroxidase and DAB, include an endogenous peroxidase block, such as 3% hydrogen peroxide for 10 minutes, and inspect a no-primary slide (standard chromogenic IHC practice). A protein block can reduce nonspecific binding; the selected image reports 3% BSA for 30 minutes (caption M08644-1). Reassess primary concentration and wash stringency if background persists while the positive-control tissue remains stained (standard IHC practice).
How should DPYSL5 staining be scored across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the eligible cell population and compartment before scoring, because HPA reports different stained cell types across tissues (HPA tissue IHC). For intensity-based scoring, assign 0–3 to each eligible cell and calculate an H-score from the percentages at each intensity, giving a 0–300 range (standard IHC scoring practice). Report the percentage of positive eligible cells alongside the H-score, and score nuclear and cytoplasmic signal separately when both occur (standard IHC practice; HPA subcellular). For spatial counts, express positive cells per mm² of viable analysed tissue (standard image-analysis practice). Normalise to the same eligible cell population or viable area across sections, and hold detection and threshold settings constant (standard IHC quantification practice).
How can I distinguish true DPYSL5 staining from chromogenic artefact?
Look for reproducible staining within identifiable cells and compare it with same-run positive and no-primary controls (standard IHC practice). Caudate neuronal cells provide a high-staining reference, whereas adipose tissue adipocytes are reported as undetected (HPA tissue IHC). A cytoplasmic pattern agrees with tissue IHC and UniProt, but nuclear staining needs independent scrutiny because HPA subcellular imaging also supports nucleoplasmic localisation (HPA tissue IHC; UniProt Q9BPU6 localisation; HPA subcellular). Discount colour confined to section edges, necrotic material or extracellular deposits when judging cellular positivity (standard IHC practice). Persistent colour in a no-primary peroxidase slide suggests endogenous enzyme or detection background rather than confirmed DPYSL5 signal (standard chromogenic IHC practice).
Boster reagents
Best DPYSL5 / Dihydropyrimidinase-related protein 5 IHC Antibodies
Catalog anti-DPYSL5 antibodies have human tonsil paraffin-section IHC and human SH-SY5Y cell IF images (catalog figure captions); all three list Human, Mouse, and Rat reactivity (catalog reactivity).
Real IHC data
Anti-DPYSL5 Antibody (C-term)
Cat # M08644-1
Real IF data
Anti-DPYSL5 Antibody (C-term)
Cat # M08644
Real IF data
Anti-DPYSL5 Antibody
Cat # M08644-2
M08644-1 has an IHC-P image of human tonsil paraffin sections (M08644-1 figure caption). M08644 and M08644-2 each have an IF image of human SH-SY5Y cells (respective figure captions).
Which to pick: For tissue IHC, choose M08644-1: its own paraffin-section caption reports citrate retrieval at pH 6 and a 1:25 primary dilution (M08644-1 figure caption). For cell IF/ICC, choose rabbit polyclonal M08644 or mouse monoclonal M08644-2; each has a cytoplasmic SH-SY5Y IF image at 1:25 (catalog antibody descriptions and respective figure captions). For Mouse or Rat samples, all three list reactivity, while the supplied images show Human samples only; select by application and validate staining in the intended species (catalog reactivity and figure captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9BPU6 (DPYL5_HUMAN, Dihydropyrimidinase-related protein 5).
- Human Protein Atlas. DPYSL5 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. DPYSL5 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. DPYSL5 antibody validation summary (2 antibodies).
- DPYSL5 is highly expressed in treatment-induced neuroendocrine prostate cancer and promotes lineage plasticity via EZH2/PRC2. Communications biology 2024 — PMC10796342.
- Differentially expressed proteins underlying childhood cortical dysplasia with epilepsy identified by iTRAQ proteomic profiling. PloS one 2017 — PMC5319751.
- PubMed PMID:11034345 — UniProt-cited evidence.
- PubMed PMID:11220734 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DPYSL5 (Dihydropyrimidinase-related protein 5). Expected localisation Cytoplasmic staining in CNS and testis (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet M08644-1). Validated IHC antibody: M08644-1 at 1:25 (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Neuronal cells); negative Adipose tissue. IHC-P workflow on paraffin sections. Protocol steps: 1. Sample: Formaldehyde-fixed, paraffin-embedded tissue sections (datasheet M08644-1; sample unspecified); 2. Fixation: Image formalin-fixed; duration unreported (datasheet M08644-1); 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: Citrate pH 6 (datasheet M08644-1); 20 min, 95–100 °C (standard); 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-DPYSL5, 1:25 (datasheet M08644-1); 9. Primary incubation: 1 hours at 37°C (datasheet M08644-1). Watch for: Fixation: Formaldehyde-fixed paraffin sections were used (selected-SKU IHC image M08644-1); Caveat: Cerebellar granular cells were undetected (HPA tissue IHC); Isoform / epitope: No annotated isoforms or processing; epitope impact unknown (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q9BPU6, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet M08644-1, PMC10796342, PMC5319751. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DPYSL5/immunohistochemistry-ihc-guide.