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- Table of Contents
DAPK1 / Death-associated protein kinase 1 · Western blot design guide
Design a Western Blot for DAPK1
Real validated DAPK1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-DAPK1 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
DAPK1 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 | ~160 kDa | |
| Observed band | ~160 kDa | |
| Gel | 5–20% (catalog A01161-1) | |
| Positive control | Esophagus (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 C7 membership | |
| Isoform | 4 isoform(s) |
Section 1
Real Curated DAPK1 Western Blot Protocols
The A01161-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat liver, mouse liver (catalog A01161-1) |
| Gel % | 5–20% (catalog A01161-1) |
| Load | 30 ug; reducing conditions (catalog A01161-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01161-1) |
| Membrane | nitrocellulose membrane (catalog A01161-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01161-1) |
| Primary antibody | A01161-1 · 0.5 μg/mL (catalog A01161-1) |
| Primary incubation | overnight at 4°C (catalog A01161-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01161-1) |
| Secondary incubation | 1.5 hour at RT (catalog A01161-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01161-1) |
| Detection | ECL (catalog A01161-1) |
Section 2
What Is the Expected DAPK1 Western Blot Band Size?
DAPK1 is predicted at 160 kDa and observed at ~160 kDa; the supplied evidence establishes no size discrepancy.
| Band at ~160 kDa | Matches the empirical DAPK1 band and predicted mass |
| Additional bands at different positions | Could reflect isoforms 1, 2, 3, or 4; their migration is not established |
| Closely spaced bands near ~160 kDa | Could reflect different phosphorylation states; visible separation is not established |
| Weak band in soluble lysate | DAPK1 may be retained in the cytoskeletal fraction |
💡Expected DAPK1 appearanceDAPK1 is predicted at 160 kDa and observed at ~160 kDa in reducing tissue lysates; confirm band identity with an appropriate control.
| UniProt predicted mass | Places the reference band near 160 kDa, consistent with the observed ~160 kDa band |
| Splice isoform 1 | May differ in size from other isoforms; its individual mass is not supplied |
| Splice isoform 2 | May differ in size from other isoforms; its individual mass is not supplied |
| Splice isoform 3 | May differ in size from other isoforms; its individual mass is not supplied |
| Splice isoform 4 | May differ in size from other isoforms; its individual mass is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytoskeleton-associated DAPK1 may be poorly recovered | Check the insoluble fraction and use a positive tissue control |
| Band higher than expected | Phosphorylation could affect migration, but a shift is not established | Compare phosphatase-treated samples and confirm identity with a second antibody |
| Band lower than expected | An alternative isoform or protein breakdown is possible; individual sizes are unknown | Compare isoform expression and confirm identity with a second antibody |
| Multiple bands | Alternative splicing or phosphorylation may contribute | Compare isoform expression, phosphatase treatment, and a DAPK1-negative control |
| Weak or no signal | Cytoskeleton-associated DAPK1 may be underrepresented in the tested fraction | Check fraction recovery and run a positive tissue control |
Experiment evidence
Western blot images showing DAPK1 band
A01161-1 · DAPK1
M01161 · DAPK1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for DAPK1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for DAPK1 in Western blot, you can use esophagus tissue lysate, which HPA scores high for DAPK1.
Positive control: Esophagus (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: Cytoplasmic DAPK1 should be accessible in tissue lysates, with HPA supporting adipose tissue as a negative control.
HPA tissue expression evidence for DAPK1
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 |
|---|---|---|---|---|
| Esophagus | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Nasopharynx | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Placenta | trophoblastic cells | High | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | High | Protein (IHC) | HPA → |
| Skin | keratinocytes | 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 → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced DAPK1 Western Blot Tips
Deeper troubleshooting and optimisation questions for DAPK1, answered from its protein features.
How should DAPK1 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 DAPK1 isoforms produce different bands?
Isoforms · Yes. Four isoforms are listed. Relative to the canonical sequence, isoform 2 lacks residues 1–446 and 784–1430 and replaces 742–783; isoform 4 lacks 805–870; isoform 3 changes the residue at 1430 to a longer ending. Check whether the antibody recognizes a region retained in the isoform of interest.
Which DAPK1 phosphorylation sites matter when interpreting a blot?
PTM · UniProt lists phosphoserines at 289 (by RPS6KA1 and RPS6KA3), 308 (by autocatalysis), 319, 333, 734 (by MAPK1), and 1115. These are UniProt coordinates; antibody or paper numbering may differ. Confirm the antibody's stated site and numbering before interpreting a phospho-specific signal.
Does this guide establish induction of DAPK1?
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 DAPK1 Western blot?
Transfer · For the approximately 160 kDa canonical protein, verify that transfer captures the high-mass band by checking the membrane and residual protein in the gel. The supplied features do not specify a transfer method or settings.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01161-1 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 should DAPK1 bands be quantified?
Quantitation · Quantify a defined band consistently across samples. Four isoforms and six listed phosphoserines make antibody target and band identity important: a total-DAPK1 signal and a site-specific phospho-DAPK1 signal answer different questions. Report which band and antibody target were measured.
Why does DAPK1 appear near 160 kDa?
Interpretation · The observed band near 160 kDa matches the supplied predicted mass of 160 kDa. The listed phosphorylation sites do not, by themselves, establish a visible shift or explain any mass difference.
First check whether the antibody's target region is retained in the relevant isoform. Isoform 2 has extensive deletions, isoform 4 lacks residues 805–870, and isoform 3 has an altered ending. The listed phosphoserines alone cannot establish that an unexpected band is a phosphorylation-dependent shift.
First check whether the antibody's target region is retained in the relevant isoform. Isoform 2 has extensive deletions, isoform 4 lacks residues 805–870, and isoform 3 has an altered ending. The listed phosphoserines alone cannot establish that an unexpected band is a phosphorylation-dependent shift.
Boster reagents
DAPK1 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-DAPK1 Antibody Picoband®
Cat # A01161-1
Real WB data
Anti-DAP Kinase 1 DAPK1 Rabbit Monoclonal Antibody
Cat # M01161
Two the supplier anti-DAPK1 antibodies list human, mouse, and rat reactivity and have Western blot images. A01161-1 shows an approximately 160 kDa band in rat and mouse liver lysates; M01161 shows DAPK1 expression in HeLa lysate. These examples do not establish performance across all specimens.
Which to pick: Choose A01161-1 for a starting point with rat or mouse liver lysates: its caption reports a 160 kDa band and detailed conditions. For HeLa lysate, M01161 has a WB image, though its caption provides fewer experimental details.
Source: BosterBio DAPK1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P53355.
- Human Protein Atlas. DAPK1 tissue expression.
- PMC11744996 — target-verified WB comparison
- PMC4047864 — target-verified WB comparison
- PMC8824877 — target-verified WB comparison