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- Table of Contents
DOCK2 / Dedicator of cytokinesis protein 2 · Western blot design guide
Design a Western Blot for DOCK2
Real validated DOCK2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-DOCK2 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
DOCK2 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 | ~211.9 kDa | |
| Gel | 4–12% gradient (standard starting point) | |
| Positive control | Appendix (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 + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated DOCK2 Western Blot Protocols
The M03836-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Jurkat cell lysate (catalog M03836-1) |
| Gel % | 4–12% gradient (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M03836-1; 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 DOCK2 Western Blot Band Size?
DOCK2 has a predicted mass of 211.9 kDa; isoforms and a probable homodimer could affect bands, but their migration is unverified.
| Band near 212 kDa | Consistent with the 211.9 kDa predicted DOCK2 mass; confirm identity with controls |
| Band near twice the predicted mass | Could reflect the probable homodimer if it survives sample preparation |
| Several bands at different positions | Could reflect isoforms 1 and 2 if they differ in migration |
| No band in whole-cell lysate | Unexpected from localization alone because DOCK2 is intracellular; check expression and detection |
💡Expected DOCK2 appearanceUniProt predicts 211.9 kDa for DOCK2, but no empirical band size is supplied; use band-identity controls before assigning a band near 212 kDa or any additional band.
| UniProt predicted mass | Places the reference full-length size at 211.9 kDa |
| Isoform 1 | May migrate differently from isoform 2; its individual mass is unspecified |
| Isoform 2 | May migrate differently from isoform 1; its individual mass is unspecified |
| Probable homodimer | Could appear near twice the monomer size if it survives sample preparation |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | DOCK2 is intracellular, so localization does not explain absence from whole-cell lysate | Check sample expression, loading, transfer, and antibody detection |
| Band higher than expected | A probable homodimer may persist during sample preparation | Compare denaturing conditions and confirm band identity |
| Band lower than expected | An alternative isoform or an unrelated band is possible; no isoform mass is supplied | Confirm identity with an independent antibody or DOCK2 depletion |
| Multiple bands | Isoforms 1 and 2 are annotated, but distinct migration is unverified | Use DOCK2 depletion and an independent antibody to identify specific bands |
| Weak or no signal | Low detectable DOCK2 in the tested sample or inefficient detection | Check loading and transfer and compare a DOCK2-positive sample |
Experiment evidence
Western blot images showing DOCK2 band
M03836-1 · DOCK2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for DOCK2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for DOCK2 in Western blot, you can use appendix tissue, which HPA scores High.
Positive control: Appendix (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: DOCK2 associates with membranes and the cytoskeleton, so adequate protein extraction may require attention.
HPA tissue expression evidence for DOCK2
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 | lymphoid tissue | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Kidney | proximal tubules (microvilli) | High | Protein (IHC) | HPA → |
| Lung | macrophages | High | Protein (IHC) | HPA → |
| Lymph node | 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 → |
| Bronchus | respiratory epithelial cells | 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 DOCK2 Western Blot Tips
Deeper troubleshooting and optimisation questions for DOCK2, answered from its protein features.
How should DOCK2 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 DOCK2 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks several stretches of the canonical sequence, including residues 956–1830, so it could produce a substantially smaller band. The supplied features do not establish either isoform’s observed migration.
Check the antibody epitope against the alternative sequence. Isoform 2 lacks canonical residues 1–449, 462–494, 630–766 and 956–1830; residues 934–955 are replaced. An epitope within a missing stretch would not detect isoform 2. These coordinates refer to the supplied canonical sequence.
Check the antibody epitope against the alternative sequence. Isoform 2 lacks canonical residues 1–449, 462–494, 630–766 and 956–1830; residues 934–955 are replaced. An epitope within a missing stretch would not detect isoform 2. These coordinates refer to the supplied canonical sequence.
Which DOCK2 phosphorylation sites matter when comparing conditions?
PTM · UniProt lists phosphoserines at canonical positions 588, 593, 1685, 1706, 1731 and 1784. Sites 588 and 593 lie outside the listed deletions in isoform 2; the four later sites lie within its missing 956–1830 stretch. Compare matched samples if probing phosphorylation, but these features specify no inducing condition or visible band shift.
N6-acetyllysine is listed at canonical positions 304 and 738. Both positions lie in regions missing from isoform 2. If probing either site, check that the antibody and sample target the canonical sequence. UniProt coordinates may differ from antibody or paper numbering; the supplied features do not establish a detectable mass shift.
N6-acetyllysine is listed at canonical positions 304 and 738. Both positions lie in regions missing from isoform 2. If probing either site, check that the antibody and sample target the canonical sequence. UniProt coordinates may differ from antibody or paper numbering; the supplied features do not establish a detectable mass shift.
Does this guide establish induction of DOCK2?
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 DOCK2 Western blot?
Transfer · At a predicted 211.9 kDa, canonical DOCK2 is large. Start with a transfer setup validated for proteins around 212 kDa and check transfer efficiency with an appropriate high molecular weight marker. The supplied features do not establish a single optimal method or transfer time.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M03836-1 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 DOCK2 localization affect quantitation?
Quantitation · DOCK2 is listed at the endomembrane system as a peripheral membrane protein and in the cytoplasm and cytoskeleton, where it colocalizes with F-actin. Keep sample preparation and the measured fraction consistent across lanes; compare matched fractions if assessing redistribution. A change in one fraction alone need not represent a change in total DOCK2.
How should the predicted DOCK2 mass guide band identification?
Interpretation · Canonical DOCK2 has a predicted mass of 211.9 kDa. Use this as a starting point when selecting a molecular weight range; no observed band position is supplied. The listed modifications alone do not establish a visible shift or explain any difference from the predicted mass.
First check band size and antibody epitope against the listed isoform 2 deletions and replacement. A probable homodimer is also listed, but these features do not establish that any particular extra band is an isoform or dimer. The phosphorylation and acetylation sites likewise do not, by themselves, explain an observed shift.
First check band size and antibody epitope against the listed isoform 2 deletions and replacement. A probable homodimer is also listed, but these features do not establish that any particular extra band is an isoform or dimer. The phosphorylation and acetylation sites likewise do not, by themselves, explain an observed shift.
Boster reagents
DOCK2 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-DOCK2 Rabbit Monoclonal Antibody
Cat # M03836-1
M03836-1 is a rabbit monoclonal anti-DOCK2 antibody listed for human reactivity. Its Western blot image shows DOCK2 expression in Jurkat cell lysate; the supplied evidence does not establish performance in other samples.
Which to pick: Only M03836-1 is listed. It has a Western blot image from Jurkat cell lysate, making it the documented option here for a human DOCK2 blot; confirm suitability for your sample and conditions.
Source: BosterBio DOCK2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q92608.
- Human Protein Atlas. DOCK2 tissue expression.
- PMC8062590 — target-verified WB comparison
- PMC6647034 — target-verified WB comparison
- PMC3178750 — target-verified WB comparison