DAZAP1 / DAZ-associated protein 1 · IHC design guide

Design Immunohistochemistry for DAZAP1

Plan DAZAP1 chromogenic IHC on paraffin sections using the nuclear tissue pattern as a reference (HPA tissue IHC). The guide covers fixation consistency, the catalog antibody's IHC conditions, and interpretation of developmental-stage differences in germ cells (UniProt).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for DAZAP1 (IHC for DAZAP1): expected localisation Predominantly nuclear in tissue sections (HPA tissue IHC), antibody A07947-1, validated IHC image, and IHC protocol steps — Printable **DAZAP1** IHC protocol sheet — expected localisation Predominantly nuclear in tissue sections (HPA tissue IHC), antibody A07947-1, controls and protocol steps. Open the full DAZAP1 IHC guide →

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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 formalin fixation consistent across sections (standard IHC practice; not target-specific)
	Caveat	Germ-cell localisation may vary by developmental stage (UniProt)
	Regulation	Spermatogenesis-associated expression (UniProt)
	Isoform / epitope	2 isoforms; epitope effects unresolved (UniProt)

Section 1

Recommended DAZAP1 IHC & IF Protocols

The catalog antibody’s IHC-P protocol is accompanied by published DAZAP1 protocols for pancreatic cancer tissues (PMC11699656) and gastric tumors and tissue microarrays (PMC12987562).

Recommended immunohistochemistry (IHC-P) protocol parameters
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 citrate pH 6 heat retrieval (datasheet A07947-1); the gastric tumor protocol specifies 95°C for 15 min (PMC12987562).

Section 2

What Is the Expected DAZAP1 Staining Pattern?

In paraffin-section IHC, expect DAZAP1 chiefly in nuclei across many cell types (HPA tissue IHC: ubiquitous nuclear expression; Enhanced reliability). High staining is reported in appendix glandular cells, bone-marrow hematopoietic cells and several epithelial and neural populations (HPA tissue IHC: High). DAZAP1 has no transmembrane segment, so a membrane-rim pattern is unexpected (UniProt Q96EP5: topology). UniProt also lists cytoplasm and nucleus, with a predominantly cytoplasmic annotation by similarity (UniProt Q96EP5: subcellular location).

What am I looking at on my slide?
Discrete nuclear chromogen in appendix glandular cells or bone-marrow hematopoietic cells.	This matches the reported high-staining populations and broad nuclear profile (HPA tissue IHC: High; ubiquitous nuclear expression). Judge intensity against a control section stained in the same run (general IHC practice).
Predominantly membrane-rim staining, with little nuclear signal in an HPA high-staining cell population.	Treat this as a suspect compartment pattern: DAZAP1 has no transmembrane segment (UniProt Q96EP5: topology), while tissue IHC is predominantly nuclear (HPA tissue IHC: ubiquitous nuclear expression). Review morphology, background and detection controls before calling it positive (general IHC practice).
Strong chromogen in smooth-muscle cells or skeletal-muscle myocytes.	These are low-staining populations in the supplied tissue survey (HPA tissue IHC: Low in smooth muscle cells and skeletal-muscle myocytes). Disproportionately strong staining could reflect cross-reactivity or endogenous detection activity; compare a no-primary control and an HPA high-staining tissue (general IHC practice).
Diffuse colour over cells and surrounding tissue, without a readable nuclear boundary.	This does not establish DAZAP1 localisation (HPA tissue IHC: ubiquitous nuclear expression). Uneven blocking, residual detection activity or excessive development can create nonspecific colour; assess a no-primary control and shorten development if indicated (general IHC practice).
No nuclear signal in appendix glandular cells or bone-marrow hematopoietic cells.	A negative result there conflicts with reported high staining (HPA tissue IHC: High in appendix glandular cells and bone-marrow hematopoietic cells). First check section integrity, control staining and the validated IHC workflow before interpreting other tissues as DAZAP1-negative (general IHC practice).

💡Expected DAZAP1 appearanceCall a section positive when cell nuclei show distinct chromogen, often strong in HPA high-staining populations such as appendix glandular cells (HPA tissue IHC: ubiquitous nuclear expression; High in appendix glandular cells); isolated membrane rims or structureless diffuse colour are suspect patterns (UniProt Q96EP5: no transmembrane segment; general IHC practice).

How each factor affects the staining
Tissue and cell selection	HPA reports high staining in appendix glandular, bone-marrow hematopoietic, bronchial respiratory epithelial, caudate glial, cerebellar granular-layer, cerebral-cortex neuronal, cervical squamous epithelial and colon glandular cells (HPA tissue IHC: High). Its low-staining examples include parathyroid glandular cells, smooth-muscle cells, skeletal-muscle myocytes and soft-tissue fibroblasts (HPA tissue IHC: Low).
Evidence and antibody validation	The HPA tissue profile is rated Enhanced for agreement between staining and RNA expression; HPA004201 and HPA004631 each have Enhanced IHC status (HPA tissue IHC: reliability; HPA antibodies: IHC Enhanced). These ratings support the observed pattern but do not define a dilution or prove every specimen will have the same intensity (HPA tissue IHC: profile; general IHC practice).
Compartment annotation	HPA tissue IHC describes ubiquitous nuclear expression, whereas UniProt records cytoplasm and nucleus and notes predominant cytoplasm by similarity (HPA tissue IHC: profile; UniProt Q96EP5: subcellular location). Interpret paraffin-section staining against the observed HPA tissue pattern; report reproducible cytoplasmic staining separately rather than forcing a single-compartment call (general IHC practice).
Isoforms and antigen identity	UniProt lists two DAZAP1 isoforms and one protein chain spanning residues 1–407, with no signal peptide or propeptide (UniProt Q96EP5: isoforms; processing). The supplied evidence gives no antibody epitope, so it cannot establish whether both isoforms are detected or predict an epitope-specific staining difference.
What should IF/ICC show?	Predominantly nucleoplasmic signal, with supported cytosolic localisation; midbody localisation is uncertain (HPA subcellular ICC-IF: main and additional locations). This describes the separate IF/ICC evidence and is not an IHC protocol option (HPA subcellular ICC-IF).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
An HPA high-staining control tissue has no signal.	A failed staining run, damaged section or weak detection is possible (general IHC practice); HPA reports high staining in appendix glandular cells and bone-marrow hematopoietic cells (HPA tissue IHC: High).	Inspect the section and run controls; check the catalog antibody's IHC-P instructions, retrieval settings and detection reagents, then repeat with a high-staining control (general IHC practice). No DAZAP1-specific retrieval requirement is supplied.
The whole section develops colour, including spaces without cells.	Non-specific deposition or endogenous detection activity may obscure the expected nuclear pattern (general IHC practice; HPA tissue IHC: ubiquitous nuclear expression).	Compare a no-primary control; verify blocking, washes and chromogen development for the chosen detection system (general IHC practice). Score DAZAP1 only where cellular boundaries and nuclei remain interpretable.
Staining is mainly on cell borders.	This disagrees with nuclear tissue IHC and lacks support from the recorded topology (HPA tissue IHC: ubiquitous nuclear expression; UniProt Q96EP5: no transmembrane segment).	Check the no-primary control and tissue morphology, then review antibody identity and detection conditions before accepting the signal (general IHC practice).
Strong colour appears chiefly in smooth muscle or myocytes.	Those populations are reported as low staining; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC: Low in smooth-muscle cells and skeletal-muscle myocytes; general IHC practice).	Compare adjacent high-staining cells where available and a no-primary control; record the cell type and compartment before changing the scoring threshold (general IHC practice).
A section shows cytoplasmic colour as well as nuclear colour.	UniProt includes cytoplasm and nucleus, while HPA paraffin tissue IHC describes a broadly nuclear pattern (UniProt Q96EP5: subcellular location; HPA tissue IHC: profile).	Score nuclear and cytoplasmic components separately, check whether cytoplasmic colour persists above the no-primary control, and document the discrepancy (general IHC practice).
A low-staining tissue looks negative.	Low staining in parathyroid glandular cells, smooth-muscle cells, skeletal-muscle myocytes or soft-tissue fibroblasts is compatible with the supplied survey (HPA tissue IHC: Low).	Confirm that an HPA high-staining control worked before reporting a tissue-level absence; a negative slide alone does not establish absent DAZAP1 protein (general IHC practice).

Source: UniProt Q96EP5 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing DAZAP1 staining

IHC analysis of DAZAP1 using anti-DAZAP1 antibody (A07947-1). DAZAP1 was detected in paraffin-embedded section of mouse testis tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-DAZAP1 Antibody (A07947-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

A07947-1 · IHCIHC analysis of DAZAP1 using anti-DAZAP1 antibody (A07947-1). DAZAP1 was detected in paraffin-embedded section of mouse testis tissues. Heat

IHC analysis of DAZAP1 using anti-DAZAP1 antibody (A07947-1). DAZAP1 was detected in paraffin-embedded section of rat testis tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-DAZAP1 Antibody (A07947-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

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.

← 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 and expect staining in glandular cells (HPA: Appendix glandular cells, High). HPA detects DAZAP1 in all 45 scored tissues, so no negative tissue is available; use no-primary and isotype controls, and expect any cells without specific signal on the positive slide to show counterstain only, with no DAB above background (HPA: no negative rows; detected in all 45 tissues).

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) slide, a host- and clonality-matched rabbit IgG isotype control, and DAZAP1 knockout material or cognate peptide competition if available (caption: rabbit primary antibody). For appendix IHC, quench endogenous peroxidase and check endogenous biotin background before biotin-based SABC/DAB detection (caption: biotinylated secondary and SABC/DAB).

⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A07947-1 paraffin-section caption does not state a fixative (caption: fixative not stated). Heat retrieval in citrate buffer at pH 6 for 20 minutes was used for mouse testis IHC, but the caption does not establish that retrieval is required (caption: mouse testis paraffin-section IHC). The evidence does not establish whether frozen sections or IF/ICC are easier; in appendix sections, assess glandular-cell staining against any luminal debris or background (HPA: Appendix glandular cells, High).

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, background and cellular localisation before comparing signal across samples.

Which retrieval condition should I try first for DAZAP1 in paraffin sections?

Start with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A07947-1). The selected paraffin-section example used this condition before overnight incubation with 1 µg/ml primary antibody at 4°C (datasheet A07947-1). If staining is weak, vary heating within a small matched series while keeping section thickness, antibody concentration and detection constant (standard IHC practice). Record tissue preservation alongside signal, because excessive heating can damage morphology and complicate compartment scoring (standard IHC practice). Judge improvement against a positive section and a no-primary control processed in the same run (standard IHC practice).

How should I troubleshoot fixation when DAZAP1 staining varies between paraffin blocks?

The selected DAZAP1 image documents paraffin-embedded mouse testis, but does not state its fixative or fixation duration (datasheet A07947-1). Target-specific sensitivity to fixation is therefore unknown; the HPA staining pattern and DAZAP1 topology do not establish a fixation condition (HPA tissue IHC; UniProt Q96EP5 topology). Compare blocks with recorded fixation histories using identical pH 6 citrate retrieval and detection settings (datasheet A07947-1; standard IHC practice). Inspect morphology and staining together, since poor preservation can make nuclear and cytoplasmic boundaries hard to score (standard IHC practice). Use an appropriately processed control section in each run to distinguish block variation from run variation (standard IHC practice).

What nuclear and cytoplasmic pattern is plausible in DAZAP1 IHC?

Score nuclear and cytoplasmic signal separately: HPA describes ubiquitous nuclear tissue staining, while its cell images support nucleoplasmic and cytosolic localisation (HPA tissue IHC; HPA subcellular). UniProt also lists both compartments and describes stage-dependent localisation during sperm development by similarity (UniProt Q96EP5 subcellular). In the selected mouse testis section, interpret staining with cell morphology rather than assuming every germ cell has the same compartment pattern (datasheet A07947-1; UniProt Q96EP5 subcellular). Treat isolated midbody signal cautiously because HPA labels that location uncertain (HPA subcellular). Compare compartment patterns with a matched control section before calling diffuse DAB deposition DAZAP1 signal (standard IHC practice).

Could isoforms or epitope accessibility explain a changed DAZAP1 staining pattern?

DAZAP1 has 2 annotated isoforms and RNA recognition motifs at residues 10–97 and 113–190 (UniProt Q96EP5 isoforms; UniProt Q96EP5 domains). The payload does not map the A07947-1 epitope, so staining cannot establish which isoform this antibody detects (datasheet A07947-1; UniProt Q96EP5 isoforms). DAZAP1 also has reported acetylated and methylated residues, but their effect on this antibody's binding is unknown (UniProt Q96EP5 modified residues). If two sections disagree, repeat the documented pH 6 citrate retrieval and compare compartment patterns under matched detection conditions (datasheet A07947-1; standard IHC practice). Avoid assigning an isoform from chromogenic intensity alone without isoform-specific validation (standard IHC practice).

How can IF help check an ambiguous DAZAP1 IHC localisation result?

Use IF as a separate localisation check, with a validated marker identifying the expected cell population in the same section (standard IF practice). Choose spectrally separated fluorophores and inspect an unstained section for tissue autofluorescence before interpreting weak DAZAP1 signal (standard IF practice). DAZAP1 has no transmembrane segment and is reported in the nucleoplasm and cytosol; permeabilisation must permit access to intracellular epitopes, including nuclear access when needed (UniProt Q96EP5 topology; HPA subcellular; standard IF practice). HPA lists ICC/IF images, but the supplied IHC caption does not establish an IF protocol for A07947-1 (HPA subcellular; datasheet A07947-1). Compare IF localisation with compartment-scored IHC rather than treating fluorescence intensity as a direct DAB equivalent (standard IHC/IF practice).

What should I check when DAZAP1 DAB staining is diffuse or widespread?

The selected workflow used 10% goat serum blocking, a biotinylated secondary antibody and avidin-biotin detection with DAB (datasheet A07947-1). Include no-primary and secondary-only controls to identify nonspecific secondary or detection signal, and assess endogenous biotin where relevant (standard IHC practice). Apply an endogenous peroxidase block before chromogenic detection, following the detection system's instructions (standard IHC practice). Widespread nuclear staining alone is not evidence of background because HPA reports ubiquitous nuclear DAZAP1 staining (HPA tissue IHC). Compare edge and damaged regions with intact central tissue, then adjust blocking or washing while preserving the documented pH 6 retrieval condition (standard IHC practice; datasheet A07947-1).

How should I quantify DAZAP1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define cell populations and score nuclear and cytoplasmic staining separately because both compartments are reported for DAZAP1 (HPA subcellular; UniProt Q96EP5 subcellular). Report the percentage of positive cells and intensity, or calculate an H-score on a 0–300 scale using a prespecified threshold (standard IHC practice). For spatial comparisons, report positive-cell density per mm² of evaluable tissue and exclude folds, edges and damaged areas consistently (standard IHC practice). Normalise counts to the number of eligible cells or evaluable tissue area, and keep imaging and DAB development settings matched (standard IHC practice). Document tissue region and compartment because testis localisation may vary with developmental stage (UniProt Q96EP5 subcellular).

How can I distinguish credible DAZAP1 staining from artefact?

Look for staining in intact cells with interpretable nuclear or cytoplasmic boundaries, then compare the pattern with a no-primary control (standard IHC practice). Nuclear signal is plausible across many tissues according to HPA, while UniProt describes developmental changes in germ-cell localisation by similarity (HPA tissue IHC; UniProt Q96EP5 subcellular). Signal confined to an unexpected compartment or cell population warrants review of morphology and antibody specificity before a biological conclusion (standard IHC practice). Discount staining concentrated at section edges, necrotic areas or regions with endogenous enzyme activity if controls show the same pattern (standard IHC practice). Interpret the selected mouse testis image within its documented paraffin, citrate-retrieval and DAB workflow (datasheet A07947-1).

Boster reagents

Best DAZAP1 / DAZ-associated protein 1 IHC Antibodies

A07947-1 has paraffin-section IHC images from mouse, rat, and human testis (catalog image captions). No IF/ICC application or image is listed (catalog applications; catalog IF images).

Real IHC data

Anti-DAZAP1 Antibody ®

Cat # A07947-1

View product →

A07947-1 will render with its mouse testis paraffin-section IHC figure (catalog image caption). The same SKU has rat and human testis paraffin-section IHC captions and lists Human, Mouse, and Rat reactivity (catalog image captions; catalog reactivity).

Which to pick: For tissue IHC, choose A07947-1, a rabbit antibody shown on paraffin-embedded testis sections; the captions do not report a fixative (catalog host; catalog image captions). There is no IF/ICC SKU to recommend because A07947-1 has no listed IF/ICC application or image (catalog applications; catalog IF images). For cross-species IHC, A07947-1 lists Human, Mouse, and Rat reactivity, with a testis IHC caption for each species (catalog reactivity; catalog image captions).

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.