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- Table of Contents
DAZAP1 / DAZ-associated protein 1 · Western blot design guide
Design a Western Blot for DAZAP1
Source-linked DAZAP1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-DAZAP1 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
DAZAP1 Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~43.4 kDa | |
| Observed band | 43 kDa | |
| Gel | 5–20% (catalog A07947-1) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Acetylated + Methylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked DAZAP1 Western Blot Protocol Options
The A07947-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human AGS, human GES-1, human HGC-27, human Hela, human Jurkat, human 293T, rat PC-12, mouse NIH/3T3 (catalog A07947-1) |
| Gel % | 5–20% (catalog A07947-1) |
| Load | 30 ug; reducing conditions (catalog A07947-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A07947-1) |
| Membrane | nitrocellulose membrane (catalog A07947-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A07947-1) |
| Primary antibody | A07947-1 · 0.5 μg/mL (catalog A07947-1) |
| Primary incubation | overnight at 4°C (catalog A07947-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A07947-1) |
| Secondary incubation | 1.5 hour at RT (catalog A07947-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A07947-1) |
| Detection | ECL (catalog A07947-1) |
Section 2
What Is the Expected DAZAP1 Western Blot Band Size?
DAZAP1 is predicted at 43.4 kDa and observed at approximately 43 kDa; its isoforms and listed modifications have no demonstrated migration effect here.
| Band at approximately 43 kDa | matches the reported DAZAP1 band and its 43.4 kDa predicted mass |
| Single sharp band near 43 kDa | consistent with DAZAP1; its listed modifications do not establish a visible shift |
| Additional band at another position | isoforms 1 and 2 are annotated, but distinct band positions are unverified |
| Weak band in a nuclear fraction | consistent with predominantly cytoplasmic localization, which varies with developmental stage |
💡Expected DAZAP1 appearanceDAZAP1 has a predicted mass of 43.4 kDa and an observed band at approximately 43 kDa; confirm band identity with antibody specificity and appropriate sample controls.
| UniProt predicted mass | 43.4 kDa predicted; the reported band is approximately 43 kDa |
| Isoform 1 | may differ in size from isoform 2, but its mass and migration are not supplied |
| Isoform 2 | may differ in size from isoform 1, but its mass and migration are not supplied |
| Alternative splicing | creates two annotated isoforms without establishing a resolvable band difference |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | DAZAP1 abundance in the tested sample is unknown | check a positive lysate and sample loading |
| Band higher than expected | the identity of a higher band is unestablished | compare with the approximately 43 kDa band and verify antibody specificity |
| Band lower than expected | an isoform is possible, but its migration is unknown | verify band identity with an independent antibody or knockdown control |
| Multiple bands | two isoforms are annotated, but distinct bands are unverified | check antibody specificity and compare samples expressing the isoforms |
| Weak or no signal | predominantly cytoplasmic localization may limit signal in a nuclear fraction | check a whole-cell or cytoplasmic fraction and verify loading |
Experiment evidence
Western blot images showing DAZAP1 band
A07947-1 · DAZAP1
M07947 · DAZAP1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for DAZAP1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for DAZAP1 in Western blot, you can use appendix tissue, which shows high HPA expression.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA lists no tissue with undetected expression, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for DAZAP1
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| 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 → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Parathyroid gland | glandular cells | Low | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Low | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced DAZAP1 Western Blot Tips
Deeper troubleshooting and optimisation questions for DAZAP1, answered from its protein features.
How should DAZAP1 band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Could DAZAP1 isoforms produce different bands?
Isoforms · UniProt lists two isoforms. In isoform 2, residues 350..407 of the canonical sequence are replaced by a shorter sequence. Their migration has not been supplied, so do not assign a band to either isoform from position alone.
Check whether the epitope lies in the shared sequence or the region changed in isoform 2. UniProt shows an alternative sequence at canonical positions 350..407; an epitope there may not recognize both isoforms.
Check whether the epitope lies in the shared sequence or the region changed in isoform 2. UniProt shows an alternative sequence at canonical positions 350..407; an epitope there may not recognize both isoforms.
Which DAZAP1 modifications matter when interpreting bands?
PTM · UniProt lists N-acetylmethionine at position 1, N6-acetyllysine at 150, and omega-N-methylarginine at 253. These are UniProt coordinates; paper or antibody numbering may differ. Their presence does not establish a detectable band shift.
Does this guide establish induction of DAZAP1?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
How should transfer be checked for DAZAP1?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A07947-1 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How can localization affect DAZAP1 quantitation?
Quantitation · DAZAP1 is listed in the cytoplasm and nucleus, with localization varying during sperm cell development. Compare matched sample types and fractions when quantifying bands, since a change in one fraction may reflect redistribution.
Should DAZAP1 migrate above its predicted mass?
Interpretation · The supplied apparent band is 43 kDa, close to the predicted 43.4 kDa. Listed modifications alone do not establish a visible shift or explain any mass difference.
Check whether the antibody recognizes both listed isoforms, especially if its epitope overlaps canonical positions 350..407. Consider the listed modifications at UniProt positions 1, 150, and 253, but do not attribute a band to an isoform or modification without supporting evidence.
Check whether the antibody recognizes both listed isoforms, especially if its epitope overlaps canonical positions 350..407. Consider the listed modifications at UniProt positions 1, 150, and 253, but do not attribute a band to an isoform or modification without supporting evidence.
Boster reagents
DAZAP1 Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-DAZAP1 Antibody Picoband®
Cat # A07947-1
Real WB data
Anti-DAZAP1 Rabbit Monoclonal Antibody
Cat # M07947
Two the supplier anti-DAZAP1 antibodies have Western blot images. A07947-1 shows a band near the expected 43 kDa in the listed human, rat, and mouse cell lysates; M07947 shows DAZAP1 expression in Jurkat lysate. The supplied evidence does not establish broader validation.
Which to pick: Choose A07947-1 when you want a documented WB example for human, rat, or mouse cell lysates and detailed assay conditions. M07947 has a WB image for Jurkat lysate, but its supplied caption gives fewer conditions.
Source: BosterBio DAZAP1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96EP5.
- Human Protein Atlas. DAZAP1 tissue expression.
- PMC11699656 — target-verified WB comparison
- PMC10386766 — target-verified WB comparison
- PMC57980 — target-verified WB comparison