This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
DLGAP5 / Disks large-associated protein 5 · Western blot design guide
Design a Western Blot for DLGAP5
Source-linked DLGAP5 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-DLGAP5 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
DLGAP5 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 | ~95.1 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Bone marrow (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Ubl conjugation | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked DLGAP5 Western Blot Protocol Options
The A30642 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | A549 cells (catalog A30642) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A30642; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected DLGAP5 Western Blot Band Size?
DLGAP5 has a predicted full-length mass of 95.1 kDa; isoforms and phosphorylation could affect bands, but altered migration is not demonstrated.
| Band near 95.1 kDa | Consistent with the predicted mass of full-length DLGAP5; identity still needs confirmation |
| Band slightly above 95.1 kDa | May reflect a modified form; phosphorylation is annotated but its effect on migration is unproven |
| Several bands near the predicted size | Could include isoforms 1, 2, and 3 or different phosphorylation states; distinct migration is unproven |
| Weak band in a soluble cytoplasmic fraction | DLGAP5 also localizes to the nucleus and mitotic spindle |
💡Expected DLGAP5 appearanceFull-length DLGAP5 has a predicted mass of 95.1 kDa; no empirical band size is supplied, and any shifted or additional bands require band-identity controls.
| Predicted full-length mass | Provides a 95.1 kDa reference, not a measured migration position |
| CDK1 phosphorylation at Ser67 | May alter apparent migration; no shift is established |
| CDK1 phosphorylation at Thr329 | May alter apparent migration; no shift is established |
| Splice isoforms 1, 2, and 3 | May differ in size, but their masses and separation on a blot are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | DLGAP5 detection in the sampled cells or cell-cycle state is uncertain | Check loading and antibody controls; compare with mitotic-enriched cells |
| Band higher than expected | Phosphorylation is annotated, but the cause of a higher band is unproven | Compare phosphatase-treated and untreated samples and confirm band identity |
| Band lower than expected | An isoform is possible, but no isoform mass is supplied | Check antibody epitope coverage and confirm the band with an independent antibody |
| Multiple bands | Isoforms or phosphorylation states are possible, without demonstrated separation | Compare phosphatase-treated samples and use an independent antibody |
| Weak or no signal | DLGAP5 localizes to nuclear and spindle compartments as well as cytoplasm | Check sample fraction and loading; compare mitotic-enriched material |
Experiment evidence
Western blot images showing DLGAP5 band
A30642 · DLGAP5
A30642 · DLGAP5
A30642 · DLGAP5
A30642 · DLGAP5
A05112 · DLGAP5
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for DLGAP5 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for DLGAP5 in Western blot, you can use bone marrow lysate.
Positive control: Bone marrow (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Signal may depend on the abundance of mitotic cells; adipose tissue offers an HPA not-detected negative control.
HPA tissue expression evidence for DLGAP5
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 |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | High | Protein (IHC) | HPA → |
| Placenta | syncytiotrophoblasts - microvilli | High | Protein (IHC) | HPA → |
| Testis | pachytene spermatocytes | High | Protein (IHC) | HPA → |
| Tonsil | germinal center cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-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 → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced DLGAP5 Western Blot Tips
Deeper troubleshooting and optimisation questions for DLGAP5, answered from its protein features.
How should DLGAP5 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.
How could DLGAP5 isoforms affect band interpretation?
Isoforms · Isoform 2 lacks canonical residues 1–81, so it may produce a smaller band. Isoform 3 has a replacement sequence at canonical residues 807–846. Check whether the antibody epitope lies in either affected region before assigning bands to isoforms.
Which DLGAP5 phosphorylation sites matter for antibody selection?
PTM · UniProt annotates CDK1 phosphorylation at canonical residues 67, 329, 401, 402, 618, 639, 642, 759 and 839, and AURKA phosphorylation at 627, 725, 757 and 830. Check whether an antibody epitope overlaps these regions. Coordinates refer to the supplied canonical sequence; antibody or paper numbering may differ.
Does this guide establish induction of DLGAP5?
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.
What transfer method to use for DLGAP5 Western blot?
Transfer · DLGAP5 has a predicted mass of 95.1 kDa. Use a transfer setup validated for proteins around that size and check transfer with a total-protein stain. The supplied features do not specify a preferred transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A30642 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 should DLGAP5 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Why might DLGAP5 migrate away from 95.1 kDa?
Interpretation · 95.1 kDa is the predicted mass; no observed band position is supplied. DLGAP5 has 24 annotated phosphorylation sites and three isoforms, but these features alone do not establish a visible shift or explain a particular apparent mass.
DLGAP5 localizes to the spindle in mitotic cells and has annotated CDK1 and AURKA phosphorylation sites. Record cell cycle conditions when comparing samples; changes in localization or phosphorylation may affect interpretation. The supplied features do not establish that total DLGAP5 abundance increases during mitosis.
Compare bands with the predicted 95.1 kDa mass, the N-terminal deletion in isoform 2, and the C-terminal replacement in isoform 3. Check the antibody epitope and sample conditions. Phosphorylation is annotated, but it cannot by itself identify an unexpected band or prove a visible shift.
DLGAP5 localizes to the spindle in mitotic cells and has annotated CDK1 and AURKA phosphorylation sites. Record cell cycle conditions when comparing samples; changes in localization or phosphorylation may affect interpretation. The supplied features do not establish that total DLGAP5 abundance increases during mitosis.
Compare bands with the predicted 95.1 kDa mass, the N-terminal deletion in isoform 2, and the C-terminal replacement in isoform 3. Check the antibody epitope and sample conditions. Phosphorylation is annotated, but it cannot by itself identify an unexpected band or prove a visible shift.
Boster reagents
DLGAP5 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-DLGAP5/Hurp Antibody
Cat # A30642
Real WB data
Anti-HURP DLGAP5 Antibody
Cat # A05112
Real WB data
Anti-DLGAP5 Antibody
Cat # A05112-1
Three catalogued anti-DLGAP5 antibodies have Western blot images: A30642 with A549 cells, A05112 with HT-29 cells, and A05112-1 with various cell lines and reported blot conditions. No publication or independent validation evidence was supplied.
Which to pick: For rat samples, consider A30642 or A05112-1; A05112 lists human and mouse reactivity. All three have WB images. Match A549 or HT-29 to the corresponding caption, or use A05112-1 when its reported dilution and blot conditions are useful.
Source: BosterBio DLGAP5 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q15398.
- Human Protein Atlas. DLGAP5 tissue expression.
- PMC13412048 — target-verified WB comparison
- PMC13244561 — target-verified WB comparison
- PMC10900829 — target-verified WB comparison