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DAZAP1 / DAZ-associated protein 1 · IHC design guide
Design Immunohistochemistry for DAZAP1
DAZAP1 shows widespread nuclear staining in tissue sections (HPA tissue IHC). This guide helps plan paraffin-section IHC with catalog antibody A07947-1 and select high-staining controls such as appendix glandular cells (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DAZAP1 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 | Predominantly nuclear in tissue sections (HPA tissue IHC) | |
| Staining pattern | Widespread nuclear staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A07947-1) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Myocytes and fibroblasts show low staining (HPA tissue IHC) | |
| Regulation | Testis-enriched expression (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended DAZAP1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A07947-1) with published DAZAP1 staining in pancreatic tissues (PMC11699656) and gastric tumor specimens (PMC12987562).
| Sample | Paraffin-embedded mouse testis tissues; fixative not specified (datasheet A07947-1) |
| Fixation | Image fixative and duration unreported (datasheet A07947-1); 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, 20 min (datasheet A07947-1) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A07947-1) |
| Primary antibody | Rabbit anti-DAZAP1, 0.5-1μg/ml (datasheet A07947-1) |
| Primary incubation | Overnight at 4 °C (datasheet A07947-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A07947-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DAZAP1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet A07947-1); the gastric tumor protocol also specifies citrate at pH 6.0 (PMC12987562).
Section 2
What Is the Expected DAZAP1 Staining Pattern?
In paraffin-section IHC, expect DAZAP1 mainly in nuclei across many cell types (HPA tissue IHC: ubiquitous nuclear expression; reliability Enhanced). HPA reports high staining in appendix glandular cells, bone marrow hematopoietic cells, and several epithelial and neural populations (HPA tissue IHC). DAZAP1 has no transmembrane segment (UniProt Q96EP5 topology). UniProt also lists cytoplasmic localization, predominantly so by similarity, creating a source-specific difference to consider when interpreting compartments (UniProt Q96EP5 subcellular location).
| Distinct nuclear staining in appendix glandular cells or bone marrow hematopoietic cells. | This matches documented high-staining populations and the broad nuclear tissue pattern (HPA tissue IHC: High; reliability Enhanced). Assess the relevant cell nuclei, since a positive tissue can also contain other structures (general IHC practice). |
| Strong cytoplasmic staining with little or no nuclear staining across the section. | Question whether the pattern represents specific IHC signal: HPA tissue IHC reports ubiquitous nuclear expression (HPA tissue IHC). Cytoplasmic localization is listed by UniProt, predominantly by similarity, so cytoplasm alone cannot establish an artefact (UniProt Q96EP5 subcellular location). |
| Strong staining concentrated in a cell population that does not fit the tissue reference. | Compare the named cell population with HPA's cell-level observations before calling it positive (HPA tissue IHC). If staining instead follows structures expected to be negative in the assay controls, consider cross-reactivity or endogenous detection activity (general IHC practice). |
| Diffuse color throughout cells, stroma, and spaces, without clear nuclear boundaries. | A diffuse pattern is difficult to score against HPA's nuclear reference (HPA tissue IHC: ubiquitous nuclear expression). Nonspecific antibody binding or detection background are possible causes; inspect controls and morphology before assigning a DAZAP1 score (general IHC practice). |
| No detectable nuclear signal in appendix glandular cells or bone marrow hematopoietic cells. | This conflicts with HPA's reported High staining in those populations (HPA tissue IHC). First verify that the expected cells are present and that the control and detection steps worked; a blank slide alone does not establish absent DAZAP1 (general IHC practice). |
💡Expected DAZAP1 appearanceCall a convincing IHC positive when identifiable cells show distinct nuclear signal, especially HPA High populations; uniform haze or control-associated color is suspect (HPA tissue IHC; general IHC practice).
| Tissue and cell context | HPA reports High staining in appendix glandular, bone marrow hematopoietic, bronchial respiratory epithelial, caudate glial, cerebellar granular-layer, cerebral cortical neuronal, cervical squamous epithelial, and colon glandular cells (HPA tissue IHC). Its listed Low populations include parathyroid glandular cells, smooth muscle cells, skeletal myocytes, and soft-tissue fibroblasts (HPA tissue IHC). |
| Source-specific localization | HPA tissue IHC describes ubiquitous nuclear expression, while UniProt lists cytoplasm and nucleus and says predominantly cytoplasmic by similarity (HPA tissue IHC; UniProt Q96EP5 subcellular location). Interpret paraffin-section staining against the tissue IHC observation; report substantial cytoplasmic staining separately rather than assuming identical compartment balance across methods (general IHC practice). |
| Validation and scope | HPA rates the tissue profile Enhanced for consistency between antibody staining and RNA expression, and lists HPA004201 and HPA004631 as IHC Enhanced (HPA tissue IHC; HPA antibodies). These summaries support the observed pattern; they do not establish a fixation effect for this target (HPA tissue IHC). |
| Protein features and isoforms | DAZAP1 has no transmembrane segment, no signal peptide or propeptide, and two reported isoforms (UniProt Q96EP5 topology, processing, isoforms). The supplied record gives no antibody epitope or isoform-specific staining data, so these features cannot predict which isoform an IHC signal represents (UniProt Q96EP5; HPA antibodies). |
| Retrieval and detection checks | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. |
| IF/ICC Q&A: where should fluorescence appear? | HPA places DAZAP1 mainly in the nucleoplasm, with supported cytosolic and uncertain midbody localization in ICC-IF (HPA subcellular). This is an IF/ICC interpretation reference, not an IHC-P protocol or a reason to require midbody staining in paraffin sections (HPA subcellular; general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected-positive cells are blank. | The sampled section may lack the relevant cells, or the staining run may have failed (general IHC practice). Appendix glandular and bone marrow hematopoietic cells are reported High (HPA tissue IHC). | Confirm cell identity and section integrity, then check the run's positive control and detection steps before interpreting DAZAP1 as absent (general IHC practice). |
| Signal appears only in cytoplasm. | The result differs from HPA's ubiquitous nuclear tissue pattern, although UniProt also lists cytoplasmic localization and calls it predominant by similarity (HPA tissue IHC; UniProt Q96EP5 subcellular location). | Compare nuclear and cytoplasmic signal in a documented High population and review controls; record the compartment observed rather than forcing a nuclear-only score (HPA tissue IHC; general IHC practice). |
| A supposedly low-staining cell type appears strongly positive. | Cell identification or nonspecific staining may explain the discrepancy; HPA lists smooth muscle cells, skeletal myocytes, and soft-tissue fibroblasts as Low (HPA tissue IHC; general IHC practice). | Recheck morphology and the exact cell population, then compare matched controls before treating the signal as evidence of expression (general IHC practice). |
| Color is diffuse across the whole section. | Background from blocking, washing, or chromogenic detection can obscure nuclei (general IHC practice). HPA's tissue reference is nuclear (HPA tissue IHC). | Inspect the negative control and staining distribution; optimize general blocking, washing, or detection conditions while retaining a documented positive tissue for comparison (general IHC practice; HPA tissue IHC). |
| Color persists in a no-primary control. | Endogenous detection activity or reagent background can generate color without primary antibody (general IHC practice). | Investigate the detection system and, where applicable, its endogenous-activity blocking step; do not score control-associated color as DAZAP1 (general IHC practice). |
| Retrieval changes the apparent staining pattern. | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. | Compare conditions on equivalent sections with the same positive and negative controls, and accept a condition only when cell morphology and interpretable nuclear staining are retained (general IHC practice; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing DAZAP1 staining
A07947-1 · IHCIHC analysis of DAZAP1 using anti-DAZAP1 antibody (A07947-1). DAZAP1 was detected in paraffin-embedded section of mouse testis tissues. Heat
A07947-1 · IHCIHC analysis of DAZAP1 using anti-DAZAP1 antibody (A07947-1). DAZAP1 was detected in paraffin-embedded section of rat testis tissues. Heat m
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000071626-DAZAP1/tissue/cerebral+cortex#imid_2276076
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000071626-DAZAP1/tissue/colon#imid_2276018
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000071626-DAZAP1/tissue/liver#imid_2276019
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000071626-DAZAP1/tissue/kidney#imid_2276012
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA004201; green = DAZAP1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA004201; green = DAZAP1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DAZAP1 IHC & IF
🧪Run appendix first; its glandular cells should stain (HPA: Appendix glandular cells, High). HPA detects DAZAP1 in all 45 scored tissues, so use no-primary and isotype controls for the negative comparison; cells without visible glandular-cell signal on the positive slide should show only background, but no specific cell type is established as DAZAP1-negative (HPA: no negative rows).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: None in HPA: DAZAP1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show DAZAP1 in A-431, U-251MG, U2OS, Sperm, KOLF2.1J, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a concentration-matched rabbit IgG isotype control, and DAZAP1 knockout material or peptide preabsorption if the immunizing peptide is available (standard IHC practice; caption: rabbit primary antibody). Check endogenous peroxidase and biotin background in appendix before interpreting DAB signal (standard IHC practice; caption: biotin-based detection with DAB).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A07947-1 paraffin-section caption does not state the fixative (caption: fixative not stated). The caption uses citrate retrieval at pH 6 for 20 minutes, but does not establish that retrieval is required (caption: heat-mediated antigen retrieval). The supplied evidence does not establish whether frozen sections or IF are easier; evaluate background from the biotin-based DAB detection on the chosen tissue (caption: biotin-based detection with DAB; standard IHC practice).
HPA tissue IHC evidence for DAZAP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: DAZAP1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced DAZAP1 IHC Tips
Troubleshoot DAZAP1 staining in paraffin sections by checking retrieval, compartment, controls and scoring before interpreting chromogenic signal.
How should I retrieve DAZAP1 in paraffin sections?
Start with heat-mediated citrate retrieval at pH 6 for 20 minutes (datasheet A07947-1). This is the retrieval used for the catalog antibody in a paraffin-embedded mouse testis section; the caption does not state the fixative (datasheet A07947-1). Keep heating, cooling and section thickness consistent across comparison slides, and include a known positive section in each run (standard IHC practice). If staining is weak, first check tissue preservation, antibody concentration and detection performance before testing an alternative retrieval condition (standard IHC practice). Compare any alternative on adjacent sections because stronger color can also reflect increased background (standard IHC practice).
Could fixation explain weak or uneven DAZAP1 staining?
DAZAP1-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not report a fixative (datasheet A07947-1). Record fixative, time in fixative, processing history and section age for each specimen, then compare like with like (standard IHC practice). In a controlled pilot, apply the same citrate pH 6, 20-minute retrieval and 1 µg/ml primary concentration to sections with documented processing (datasheet A07947-1; standard IHC practice). Examine morphology alongside staining, since poor preservation can make compartment assignment unreliable (standard IHC practice). Neither tissue expression patterns nor DAZAP1 modifications establish a fixation effect for this antibody (HPA tissue IHC; UniProt Q96EP5).
Should DAZAP1 staining be nuclear or cytoplasmic?
Assess nuclear and cytoplasmic signal separately: tissue IHC reports ubiquitous nuclear expression, while subcellular imaging supports nucleoplasm and cytosol (HPA tissue IHC; HPA subcellular). UniProt describes DAZAP1 as predominantly cytoplasmic by similarity and reports developmental shifts between nuclei and cytoplasm in spermatogenic cells (UniProt Q96EP5). In testis, record the cell stage and compartment before comparing fields; midpachytene spermatocytes, round spermatids and elongated spermatids have different reported patterns (UniProt Q96EP5). Use a nuclear counterstain to assign chromogenic signal to individual cells, and score ambiguous overlap separately (standard IHC practice). Treat isolated midbody staining cautiously because that HPA assignment is uncertain (HPA subcellular).
Can isoforms or epitope masking change the staining pattern?
DAZAP1 has two annotated isoforms, but the supplied antibody evidence does not map its epitope or establish isoform coverage (UniProt Q96EP5; datasheet A07947-1). Its RNA recognition motifs span residues 10–97 and 113–190, and recorded modifications include acetylation at residue 150 and methylation at residue 253 (UniProt Q96EP5). These annotations identify questions for epitope review; they do not show that a modification or isoform blocks this antibody (UniProt Q96EP5; datasheet A07947-1). If staining varies unexpectedly, obtain the epitope specification and compare controls processed in the same run (standard IHC practice). Avoid assigning an isoform from chromogenic intensity alone (standard IHC practice).
How can I check DAZAP1 localisation by multiplex IF?
For an IF comparison, pair DAZAP1 with an independently validated marker of the expected spermatogenic cell population and a nuclear counterstain (UniProt Q96EP5; standard IF practice). Choose a spectrally separated red or far-red fluorophore after inspecting tissue autofluorescence and single-label controls (standard IF practice). DAZAP1 has no transmembrane segment and is reported in intracellular nuclear and cytosolic compartments, so assess permeabilisation for access to those compartments (UniProt Q96EP5; HPA subcellular; standard IF practice). Check whether the signal tracks the marked cells and their nuclei across optical planes (standard IF practice). The selected IHC caption supplies no IF fixation or permeabilisation conditions (datasheet A07947-1).
How do I reduce diffuse or misleading DAB background?
The catalog example used 10% goat-serum block, 1 µg/ml primary overnight at 4°C, and biotin-based detection with DAB (datasheet A07947-1). Start by comparing a no-primary control with the stained section, then inspect whether haze follows tissue folds, edges or damaged areas (standard IHC practice). Block endogenous peroxidase before DAB development and evaluate endogenous biotin when using biotin-based amplification (standard IHC practice). If background persists, titrate primary concentration and detection development while keeping retrieval at citrate pH 6 constant during that comparison (datasheet A07947-1; standard IHC practice). Judge improvement by retained cellular localisation as well as lower diffuse color (standard IHC practice).
What is a defensible way to quantify DAZAP1 IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue region, cell population and nuclear versus cytoplasmic compartment before scoring (standard IHC practice). Report the percentage of positive cells and an H-score from 0–300, or count positive cells per mm² when cell density is the question (standard IHC practice). Normalise counts to the number of evaluable cells or measured tissue area, and apply one threshold and imaging setup across the comparison (standard IHC practice). Record nuclear and cytoplasmic scores separately because both compartments are reported for DAZAP1 (HPA subcellular; UniProt Q96EP5). Exclude folds, necrosis and missing tissue using prespecified rules, and retain control-slide results with each scoring batch (standard IHC practice).
How can I distinguish true DAZAP1 staining from artefact?
Look for reproducible cellular staining with a plausible compartment: HPA supports nucleoplasm and cytosol, and reports widespread nuclear tissue expression (HPA subcellular; HPA tissue IHC). In testis, compare the stained cell type and developmental stage with the reported shifts among spermatocytes and spermatids (UniProt Q96EP5). Treat color confined to cut edges, folds, necrotic areas or a no-primary control as suspect, and check the endogenous peroxidase block when DAB appears nonspecific (standard IHC practice). A signal exclusively in an unexpected compartment needs confirmation on an independently processed section (standard IHC practice). Do not infer isoform identity or fixation sensitivity from staining pattern alone (UniProt Q96EP5; datasheet A07947-1).
Boster reagents
Best DAZAP1 / DAZ-associated protein 1 IHC Antibodies
The catalog antibody has real paraffin-section IHC images from human, mouse and rat testis (catalog IHC captions). No IF/ICC data are listed (catalog applications; catalog IF images).
Real IHC data
Anti-DAZAP1 Antibody ®
Cat # A07947-1
A07947-1 will render with an IHC figure from paraffin-embedded mouse testis (catalog IHC caption). Its additional IHC captions show rat and human testis; its listed applications are ELISA, IHC and WB (catalog IHC captions; catalog applications).
Which to pick: Choose A07947-1 for paraffin-section IHC, starting at 0.5–1 μg/ml (datasheet: IHC dilution); its mouse testis image used citrate retrieval at pH 6 for 20 minutes (catalog mouse IHC caption). The same SKU has human, mouse and rat IHC testis images, making it the documented cross-species choice here; the captions identify paraffin sections but do not report the fixative (catalog IHC captions). No SKU in this payload is listed for IF/ICC, so there is no documented IF/ICC pick (catalog applications; catalog IF images).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96EP5 (DAZP1_HUMAN, DAZ-associated protein 1).
- Human Protein Atlas. DAZAP1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. DAZAP1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol and midbody..
- Human Protein Atlas. DAZAP1 antibody validation summary (2 antibodies).
- RNA-binding protein DAZAP1 accelerates the advancement of pancreatic cancer by inhibiting ferroptosis. European journal of medical research 2025 — PMC11699656.
- RNA‑binding protein DAZAP1 promotes gastric cancer metastasis by enhancing NOTCH1 and JAG1 mRNA stability. International journal of oncology 2026 — PMC12987562.
- PubMed PMID:10857750 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DAZAP1 (DAZ-associated protein 1). Expected localisation Predominantly nuclear in tissue sections (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A07947-1). Validated IHC antibody: A07947-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse testis tissues; fixative not specified (datasheet A07947-1); 2. Fixation: Image fixative and duration unreported (datasheet A07947-1); 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 , 20 min (datasheet A07947-1); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A07947-1); 8. Primary antibody: Rabbit anti-DAZAP1, 0.5-1μg/ml (datasheet A07947-1); 9. Primary incubation: Overnight at 4 °C (datasheet A07947-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Myocytes and fibroblasts show low staining (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unknown (UniProt); Regulation: Testis-enriched expression (UniProt). Evidence: UniProt Q96EP5, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A07947-1, PMC11699656, PMC12987562. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DAZAP1/immunohistochemistry-ihc-guide.