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- Table of Contents
MOBP / Myelin-associated oligodendrocyte basic protein · Western blot design guide
Design a Western Blot for MOBP
Source-linked MOBP Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MOBP 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
MOBP 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 | ~21 kDa | |
| Observed band | ~21 kDa | |
| Gel | 5–20% (catalog A09516-2) | |
| Positive control | Cerebral cortex (IHC candidate; verify WB) | |
| Negative control | Adipose tissue (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 C7 membership | |
| Isoform | 4 isoform(s) |
Section 1
Source-Linked MOBP Western Blot Protocol Options
The A09516-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HepG2, human Caco-2, human K562, human SH-SY5Y, rat brain, rat C6, mouse brain, mouse Neuro-2a (catalog A09516-2) |
| Gel % | 5–20% (catalog A09516-2) |
| Load | 30 ug; reducing conditions (catalog A09516-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A09516-2) |
| Membrane | nitrocellulose membrane (catalog A09516-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A09516-2) |
| Primary antibody | A09516-2 · 0.5 μg/mL (catalog A09516-2) |
| Primary incubation | overnight at 4°C (catalog A09516-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A09516-2) |
| Secondary incubation | 1.5 hour at RT (catalog A09516-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A09516-2) |
| Detection | ECL (catalog A09516-2) |
Section 2
What Is the Expected MOBP Western Blot Band Size?
MOBP is predicted at 21 kDa and observed at ~21 kDa; isoforms and phosphorylation are listed, but their effects on migration are unestablished.
| Band at ~21 kDa | Matches the predicted mass and the empirical MOBP band. |
| Single sharp band near 21 kDa | Consistent with MOBP; confirm identity with a control. |
| Multiple discrete bands | Could reflect isoforms 1, 2, 3, or 4; distinct migration is unverified. |
| Slightly shifted band near 21 kDa | Could reflect phosphorylation at Ser99 or Ser109; a visible shift is unverified. |
💡Expected MOBP appearanceMOBP has a predicted mass of 21 kDa and an empirical band at ~21 kDa; confirm band identity with an appropriate control because isoform and phosphorylation effects on migration are unestablished.
| Predicted MOBP mass | 21 kDa, matching the empirical band at ~21 kDa. |
| Isoform 1 | Its mass relative to other isoforms is unspecified. |
| Isoform 2 | Its mass relative to other isoforms is unspecified. |
| Isoform 3 | Its mass relative to other isoforms is unspecified. |
| Isoform 4 | Its mass relative to other isoforms is unspecified. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | MOBP abundance may be low in the tested sample. | Check a brain tissue lysate positive control and sample loading. |
| Band higher than expected | A MOBP isoform or phosphorylation could affect migration, but neither shift is established. | Compare with the ~21 kDa control band and verify identity with a second antibody. |
| Band lower than expected | An alternative isoform or protein degradation is possible; its identity is unverified. | Check sample integrity and verify the band with a second antibody. |
| Multiple bands | Four MOBP isoforms are listed, but their band positions are unknown. | Compare across samples and confirm band identities with isoform-specific controls. |
| Broad smear instead of sharp band | Sample degradation or nonspecific signal is possible; no glycosylation site supports a glycoform smear. | Check sample integrity and antibody specificity. |
| Fragments below expected size | Protein degradation is possible; no cleavage feature is listed. | Prepare fresh lysate with protease inhibitors and compare with a control. |
Experiment evidence
Western blot images showing MOBP band
A09516-2 · MOBP
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MOBP Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MOBP in Western blot, you can use cerebral cortex lysate.
Positive control: Cerebral cortex (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: MOBP is present in CNS myelin, so a myelin-rich cerebral cortex sample may give a stronger signal than bulk tissue lysate.
HPA tissue expression evidence for MOBP
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 |
|---|---|---|---|---|
| Cerebral cortex | neuropil | 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 → |
| Adrenal gland | glandular cells | 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 → |
Section 3
Advanced MOBP Western Blot Tips
Deeper troubleshooting and optimisation questions for MOBP, answered from its protein features.
How should MOBP 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.
Which MOBP isoforms might produce different bands?
Isoforms · UniProt lists four isoforms. Isoform 3 substitutes residues 70–81 and lacks residues 82–183; isoform 4 changes residue 68 to a longer sequence. Isoforms 2 and 4 lack residue 183. These differences could affect migration, but band positions must be established experimentally.
Map the antibody epitope against each isoform. An epitope within canonical residues 82–183 is absent from isoform 3, while sequences near residue 68 or residues 70–81 differ in isoforms 4 and 3, respectively. Check the actual isoform sequences before treating bands as comparable.
Map the antibody epitope against each isoform. An epitope within canonical residues 82–183 is absent from isoform 3, while sequences near residue 68 or residues 70–81 differ in isoforms 4 and 3, respectively. Check the actual isoform sequences before treating bands as comparable.
Which MOBP phosphorylation sites matter for band interpretation?
PTM · UniProt annotates phosphoserines at positions 99 and 109 in the supplied sequence. These are UniProt coordinates; antibody or paper numbering may differ. Isoform 3 lacks canonical residues 82–183, so it lacks those canonical sites.
No. The annotations at Ser99 and Ser109 establish sites, not phosphorylation levels in a given sample or a detectable mobility change. If a shifted band appears, compare untreated and phosphatase-treated samples before assigning it to phosphorylation.
No. The annotations at Ser99 and Ser109 establish sites, not phosphorylation levels in a given sample or a detectable mobility change. If a shifted band appears, compare untreated and phosphatase-treated samples before assigning it to phosphorylation.
Does this guide establish induction of MOBP?
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 MOBP?
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 A09516-2 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 MOBP bands be quantified across samples?
Quantitation · Use the same band definition across samples and check whether the antibody recognizes all relevant isoforms. Isoform 3 lacks residues 82–183, and isoforms 2 and 4 lack residue 183; epitope coverage can therefore affect what the blot measures. Quantify distinct bands separately until their identities are established.
Should MOBP run at its predicted 21 kDa?
Interpretation · The supplied observed band is about 21 kDa, matching the predicted mass. MOBP also has sequence variants and two annotated phosphorylation sites, but those features alone do not establish a visible shift or explain any mass discrepancy.
First compare them with the four annotated isoforms, especially the large deletion in isoform 3 and the longer replacement sequence in isoform 4. Consider phosphorylation at UniProt Ser99 and Ser109, but do not assign a band from size alone. The supplied features list no glycosylation sites, signal peptide, or propeptide.
First compare them with the four annotated isoforms, especially the large deletion in isoform 3 and the longer replacement sequence in isoform 4. Consider phosphorylation at UniProt Ser99 and Ser109, but do not assign a band from size alone. The supplied features list no glycosylation sites, signal peptide, or propeptide.
Boster reagents
MOBP 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-MOBP Antibody Picoband®
Cat # A09516-2
The catalog reports one anti-MOBP antibody, A09516-2, with stated human, mouse, and rat reactivity. Its WB image shows an approximately 21 kDa band in the human cell, rat cell and brain, and mouse cell and brain lysates named in the caption. No publication evidence is supplied.
Which to pick: A09516-2 is the only listed option and has a WB image. The caption includes human HepG2, Caco-2, K562, and SH-SY5Y cells; rat brain and C6 cells; and mouse brain and Neuro-2a cells. Use those tested contexts to judge fit for your sample.
Source: BosterBio MOBP gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q13875.
- Human Protein Atlas. MOBP tissue expression.
- PMC12317959 — target-verified WB comparison
- PMC11849431 — target-verified WB comparison
- PMC8219819 — target-verified WB comparison