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- Table of Contents
DLK1 / Protein delta homolog 1 · Western blot design guide
Design a Western Blot for DLK1
Real validated DLK1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-DLK1 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
DLK1 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~41.3 kDa | |
| Observed band | ≈41 kDa | |
| Gel | 10% (catalog A00513-5) | |
| Positive control | Adrenal gland (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 | Glycosylated + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated DLK1 Western Blot Protocols
The A00513-5 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human A549, human MCF-7 (catalog A00513-5) |
| Gel % | 10% (catalog A00513-5) |
| Load | 30 ug; reducing conditions (catalog A00513-5) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A00513-5) |
| Membrane | nitrocellulose membrane (catalog A00513-5) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A00513-5) |
| Primary antibody | A00513-5 · 0.5 μg/mL (catalog A00513-5) |
| Primary incubation | overnight at 4°C (catalog A00513-5) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A00513-5) |
| Secondary incubation | 1.5 hour at RT (catalog A00513-5) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A00513-5) |
| Detection | ECL (catalog A00513-5) |
Section 2
What Is the Expected DLK1 Western Blot Band Size?
DLK1 is predicted at 41.3 kDa and observed near 41 kDa in reducing lysates; the small difference has no established cause.
| Band near 41 kDa | Matches the reported band in reducing whole-cell lysates and is close to the 41.3 kDa predicted mass; confirm identity with controls |
| Band above 41 kDa | Could reflect glycosylation, although its effect on migration is not established |
| Band below 41 kDa | Could reflect signal-peptide cleavage or the Short isoform; neither band position is established |
| Several bands | Long and Short isoforms are annotated, but distinct resolvable bands are not established |
| Broad smear | Variable glycosylation is possible across annotated sites; a visible smear is not established |
💡Expected DLK1 appearanceDLK1 has a predicted mass of 41.3 kDa and an empirical band near 41 kDa in reducing whole-cell lysates; confirm band identity with appropriate antibody and sample controls.
| UniProt predicted mass | 41.3 kDa provides the sequence-based reference; the reported band is approximately 41 kDa |
| O-linked GalNAc site at Ser94 | May affect apparent migration, but a measurable shift is not established |
| N-linked GlcNAc site at Asn100 | May affect apparent migration, but its size effect is not supplied |
| Partial atypical N-linked GlcNAc site at Asn165 | May contribute to variation in migration; a visible effect is not established |
| Signal peptide at residues 1–23 | Cleavage makes the mature chain shorter than the full-length precursor; its band position is not supplied |
| Long and Short isoforms | May differ in size; individual masses and resolvable migration differences are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | DLK1 is a single-pass membrane protein that may be poorly recovered | Check membrane-protein extraction and include a positive-control lysate |
| Band higher than expected | Annotated N- and O-linked glycosylation may affect migration | Compare with a validated deglycosylation control and confirm band identity |
| Band lower than expected | Signal-peptide cleavage or the Short isoform could alter size | Check antibody epitope coverage and compare with isoform-specific controls |
| Broad smear instead of sharp band | Variation across annotated glycosylation sites is possible | Check sample preparation and compare treated and untreated samples with a band-identity control |
| Multiple bands | Long and Short isoforms are annotated, though distinct bands are unproven | Compare isoform-specific controls and verify which bands the antibody recognizes |
| Weak or no signal | Membrane localization may reduce recovery in the tested preparation | Check extraction efficiency and use a positive-control lysate |
Experiment evidence
Western blot images showing DLK1 band
A00513-5 · DLK1
A00513 · DLK1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for DLK1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for DLK1 in Western blot, you can use adrenal gland lysate, where HPA reports high expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: Because DLK1 is a membrane protein, use a lysate that retains membrane proteins.
HPA tissue expression evidence for DLK1
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Colon | peripheral nerve/ganglion | Medium | Protein (IHC) | HPA → |
| Nasopharynx | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Ovary | follicle cells | Medium | 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced DLK1 Western Blot Tips
Deeper troubleshooting and optimisation questions for DLK1, answered from its protein features.
How should DLK1 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 DLK1 isoforms affect band interpretation?
Isoforms · UniProt lists Long and Short isoforms. Short lacks residues 229–301 of the canonical sequence, so an additional band could reflect an isoform. Check whether the antibody recognizes a region retained in Short before assigning the band.
Which DLK1 glycosylation sites matter when assessing band patterns?
PTM · UniProt lists O-linked sites at 94, 143, 163, 214, 222, 251, 256 and 260, and N-linked sites at 100, 165 and 172. Sites 163, 165, 172, 222, 251 and 260 are marked partial; 165 and 172 are also atypical. These are canonical-sequence coordinates, which may differ from antibody or paper numbering.
Glycosylation is a plausible consideration because DLK1 has 11 listed sites, including partially occupied sites. Site annotations alone cannot establish that a particular band represents a glycoform. Compare the bands with an appropriate glycosylation assessment before assigning them.
Glycosylation is a plausible consideration because DLK1 has 11 listed sites, including partially occupied sites. Site annotations alone cannot establish that a particular band represents a glycoform. Compare the bands with an appropriate glycosylation assessment before assigning them.
Does this guide establish induction of DLK1?
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 DLK1 Western blot?
Transfer · UniProt identifies DLK1 as a single-pass type I membrane protein but does not specify a transfer method. Check transfer efficiency around the reported approximately 41 kDa band and optimize transfer conditions empirically.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A00513-5 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 DLK1 bands be quantified?
Quantitation · Define which band or bands you are measuring and apply that choice consistently. The reported band is approximately 41 kDa, while UniProt lists two isoforms and multiple glycosylation sites. Those features make it important to distinguish a single band measurement from a combined DLK1 signal.
Why is the observed DLK1 band near its predicted mass?
Interpretation · The reported band is approximately 41 kDa, close to the 41.3 kDa predicted mass. DLK1 has a signal peptide at residues 1–23 and 11 listed glycosylation sites, but those features alone do not establish a visible shift or explain the apparent mass.
The Short isoform lacks canonical residues 229–301, making isoform expression one possibility. That deletion also removes listed glycosylation sites 251, 256 and 260. An unexpected band cannot be assigned to Short from its position alone; check antibody epitope coverage and isoform expression.
The Short isoform lacks canonical residues 229–301, making isoform expression one possibility. That deletion also removes listed glycosylation sites 251, 256 and 260. An unexpected band cannot be assigned to Short from its position alone; check antibody epitope coverage and isoform expression.
Boster reagents
DLK1 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-DLK-1/DLK1 Antibody Picoband®
Cat # A00513-5
Real WB data
Anti-Protein delta homolog 1 DLK1 Antibody
Cat # A00513
Two anti-DLK1 antibodies have WB images. A00513-5 shows an approximately 41 kDa band in human A549 and MCF-7 whole cell lysates. A00513 has a WB caption describing HepG2 in 293 cell lysate; the supplied evidence does not establish WB performance across its full stated reactivity.
Which to pick: For human A549 or MCF-7 lysates, A00513-5 has the more detailed WB example. A00513 lists human, mouse, and rat reactivity and has a WB image, but its caption does not demonstrate mouse or rat samples. Choose based on your sample species and the reported conditions.
Source: BosterBio DLK1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P80370.
- Human Protein Atlas. DLK1 tissue expression.
- PMC3338567 — target-verified WB comparison
- PMC6911156 — target-verified WB comparison
- PMC6000581 — target-verified WB comparison