This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
MARK2 / Serine/threonine-protein kinase MARK2 · Western blot design guide
Design a Western Blot for MARK2
Real validated MARK2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MARK2 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
MARK2 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 | ~87.9 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Bone marrow (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 16 isoform(s) |
Section 1
Real Curated MARK2 Western Blot Protocols
The M02117-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Neuro-2a cell lysate (catalog M02117-1) |
| 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 | M02117-1; 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 MARK2 Western Blot Band Size?
MARK2 is predicted at 87.9 kDa; isoforms, phosphorylation, and homodimerization may affect bands, but their migration effects are not demonstrated here.
| Band near 87.9 kDa | Consistent with the predicted MARK2 mass; confirm identity with antibody controls. |
| Band near twice the predicted mass | Could reflect a homodimer that persists during sample preparation. |
| Several bands near the expected region | Could reflect MARK2 isoforms, although their migration differences are unknown. |
| Close doublet near the expected region | Could reflect different phosphorylation states; the sites alone do not establish a visible doublet. |
💡Expected MARK2 appearanceMARK2 has a predicted mass of 87.9 kDa, but no empirical band size is supplied; use antibody specificity controls to identify the band because isoforms, phosphorylation, and homodimerization do not establish its migration.
| UniProt predicted mass | Places the predicted MARK2 monomer near 87.9 kDa. |
| Homodimer formation | Could yield a band near twice the monomer mass if the dimer persists during preparation. |
| Splice isoforms 1 through 8 | May differ in apparent size; individual masses and migration are not supplied. |
| Splice isoforms 9 through 16 | May differ in apparent size; individual masses and migration are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | A MARK2 homodimer may persist during preparation. | Compare reduced and non-reduced samples and verify band identity with antibody controls. |
| Band lower than expected | An alternative MARK2 isoform is possible; its size is unknown. | Check which isoforms the antibody recognizes and confirm the band with an independent antibody. |
| Multiple bands | Alternative isoforms or phosphorylation states may contribute. | Check isoform recognition and compare samples with and without phosphatase treatment. |
| Broad smear instead of sharp band | Variable phosphorylation is possible, but the cause is unestablished. | Compare phosphatase-treated samples and check sample quality. |
| Weak or no signal | Extraction may recover little membrane- or cytoskeleton-associated MARK2. | Check extraction conditions and include a positive lysate control. |
| Fragments below expected size | Sample degradation is possible; no MARK2 cleavage product is specified. | Prepare fresh lysate with protease inhibitors and verify fragments with an independent antibody. |
Experiment evidence
Western blot images showing MARK2 band
M02117-1 · MARK2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MARK2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MARK2 in Western blot, you can use appendix tissue, which HPA scores as high expression.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Bone marrow (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: MARK2 is intracellular, and HPA identifies both high-expression and not-detected tissues for controls.
HPA tissue expression evidence for MARK2
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 → |
| Cerebellum | cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuropil | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Lung | alveolar cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MARK2 Western Blot Tips
Deeper troubleshooting and optimisation questions for MARK2, answered from its protein features.
How should MARK2 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 MARK2 isoforms produce multiple bands?
Isoforms · UniProt lists 16 isoforms with several missing sequence segments. For example, isoforms 2, 3, 6, 7, 10, 12, 13 and 14 lack canonical residues 1–33. Isoform expression and band positions are not supplied, so assign bands to isoforms only with supporting evidence.
Check the antibody epitope against the listed sequence differences. Canonical residues 505–558 are missing in isoforms 2, 4, 5, 7, 9, 10, 13, 15 and 16. An antibody targeting that segment may miss those isoforms. UniProt coordinates refer to the canonical sequence; antibody numbering may differ.
Check the antibody epitope against the listed sequence differences. Canonical residues 505–558 are missing in isoforms 2, 4, 5, 7, 9, 10, 13, 15 and 16. An antibody targeting that segment may miss those isoforms. UniProt coordinates refer to the canonical sequence; antibody numbering may differ.
Which MARK2 phosphorylation sites could a phospho antibody assess?
PTM · Listed sites include canonical Thr-208, phosphorylated by LKB1 and TAOK1; Ser-212, by GSK3-beta; Ser-91, Ser-92, Ser-93 and Thr-294, by CaMK1; and Thr-596, by PKC/PRKCZ. Match the antibody’s stated site and numbering convention to the UniProt canonical coordinates before interpreting its signal.
No visible shift is established by the feature list. MARK2 has 21 listed modified residues, including autocatalytic Thr-58, Ser-274 and Thr-275. If mobility changes, compare the same isoform across conditions and assess phosphorylation separately; the listed sites alone cannot explain a measured mass difference.
No visible shift is established by the feature list. MARK2 has 21 listed modified residues, including autocatalytic Thr-58, Ser-274 and Thr-275. If mobility changes, compare the same isoform across conditions and assess phosphorylation separately; the listed sites alone cannot explain a measured mass difference.
Does this guide establish induction of MARK2?
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 MARK2?
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 M02117-1 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 MARK2 signal be quantified across cell fractions?
Quantitation · MARK2 is listed in the cytoplasm, cytoskeleton and at cell membranes, including the lateral membrane. Define whether the measurement concerns total protein or a particular fraction, and compare like fractions between samples. A fraction-specific signal may change without establishing a change in total MARK2 abundance.
Should MARK2 run at its predicted 87.9 kDa?
Interpretation · The canonical sequence has a predicted mass of 87.9 kDa. No observed band position is supplied, so this does not establish where MARK2 runs on a blot. Consider the listed isoforms and phosphorylation sites when assessing a different apparent mass; their presence alone does not demonstrate a visible shift.
Check whether the antibody epitope is present in candidate isoforms and consider the listed sequence deletions, including residues 505–558 and 644–652. MARK2 is also phosphorylated at multiple sites. These features suggest possibilities to investigate, but neither isoform identity nor a phosphorylation-dependent shift can be assigned from band position alone.
Check whether the antibody epitope is present in candidate isoforms and consider the listed sequence deletions, including residues 505–558 and 644–652. MARK2 is also phosphorylated at multiple sites. These features suggest possibilities to investigate, but neither isoform identity nor a phosphorylation-dependent shift can be assigned from band position alone.
Boster reagents
MARK2 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-MARK2 Rabbit Monoclonal Antibody
Cat # M02117-1
The catalog reports one anti-MARK2 antibody for Western blotting: rabbit monoclonal M02117-1. It is listed as reactive with human, mouse, and rat, and its WB image shows MARK2 expression in Neuro-2a cell lysate. The supplied evidence does not document other tested samples.
Which to pick: M02117-1 is the only listed option. Its reported reactivity covers human, mouse, and rat, and it has a WB image from Neuro-2a cell lysate. Choose it with that demonstrated sample context in mind.
Source: BosterBio MARK2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q7KZI7.
- Human Protein Atlas. MARK2 tissue expression.
- PMC9354676 — target-verified WB comparison
- PMC11568763 — target-verified WB comparison
- PMC3242779 — target-verified WB comparison