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- Table of Contents
MSI2 / RNA-binding protein Musashi homolog 2 · Western blot design guide
Design a Western Blot for MSI2
Source-linked MSI2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MSI2 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
MSI2 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 | ~35.2 kDa | |
| Observed band | ~35 kDa | |
| Gel | 10% (catalog A02898-2) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked MSI2 Western Blot Protocol Options
The A02898-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human SH-SY5Y, human RT4, human U251, human HepG2, rat brain, rat C6, mouse brain, mouse Neuro-2a (catalog A02898-2) |
| Gel % | 10% (catalog A02898-2) |
| Load | 30 ug; reducing conditions (catalog A02898-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02898-2) |
| Membrane | nitrocellulose membrane (catalog A02898-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02898-2) |
| Primary antibody | A02898-2 · 0.5 μg/mL (catalog A02898-2) |
| Primary incubation | overnight at 4°C (catalog A02898-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02898-2) |
| Secondary incubation | 1.5 hour at RT (catalog A02898-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02898-2) |
| Detection | ECL (catalog A02898-2) |
Section 2
What Is the Expected MSI2 Western Blot Band Size?
MSI2 is predicted at 35.2 kDa and observed at ~35 kDa; the small difference has no established cause, and band identity needs controls.
| Band at ~35 kDa | Matches the empirical MSI2 band and its 35.2 kDa predicted mass; confirm identity with controls |
| Additional bands at different positions | Could reflect isoforms 1, 2, or 3; their migration is not established |
| Close doublet near 35 kDa | Could reflect phosphorylation, but a visible shift has not been demonstrated |
| Faint ~35 kDa band in whole-cell lysate | Cytoplasmic MSI2 may be poorly detected in this preparation |
💡Expected MSI2 appearanceMSI2 has a predicted mass of 35.2 kDa and an empirical band at ~35 kDa; confirm band identity with a positive control or MSI2 depletion.
| Predicted MSI2 mass | 35.2 kDa predicts migration near the empirical ~35 kDa band |
| Isoforms 1, 2, and 3 | May differ in size, but their masses and resolvable migration are not supplied |
| Phosphoserine at position 6 and annotated position 261 | May alter mobility; no visible shift is established |
| N-acetylmethionine at position 1 | Adds a small modification without an established visible shift |
| Omega-N-methylarginine at position 228 and annotated position 261 | Adds small modifications without an established visible shift |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytoplasmic MSI2 may be poorly recovered or detected | Check cytoplasmic extraction and run a positive-control lysate |
| Band higher than expected | A different isoform or modified MSI2 is possible; the cause is unproven | Compare with a positive control and verify identity by MSI2 depletion |
| Band lower than expected | An alternative isoform or degradation is possible; neither migration is established | Use MSI2 depletion and check sample handling |
| Multiple bands | Isoforms 1, 2, and 3 or modified MSI2 may contribute | Check band identity by MSI2 depletion; compare isoform-specific expression if available |
| Weak or no signal | MSI2 detection in this preparation may be low | Verify loading, transfer, and signal with a positive-control lysate |
| Fragments below expected size | MSI2 degradation is possible given its reported ubiquitination and degradation | Prepare fresh lysate with protease inhibitors and verify fragments by MSI2 depletion |
Experiment evidence
Western blot images showing MSI2 band
A02898-2 · MSI2
M02898 · MSI2
A02898 · MSI2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MSI2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MSI2 in Western blot, you can use adrenal gland tissue, which shows high HPA expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: No HPA tissue is listed as not detected, so use siRNA knockdown or a KO line for a negative control.
HPA tissue expression evidence for MSI2
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 → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Caudate | neuronal cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | glial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Lymph node | non-germinal center cells | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Medium | Protein (IHC) | HPA → |
Section 3
Advanced MSI2 Western Blot Tips
Deeper troubleshooting and optimisation questions for MSI2, answered from its protein features.
How should MSI2 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 MSI2 isoforms produce different bands?
Isoforms · Yes. UniProt lists three isoforms. Isoform 3 lacks canonical residues 1–104; isoforms 2 and 3 replace residues 243–255 and lack residues 256–328. Isoform 2 also replaces residues 1–21. Compare the detected band and antibody epitope with these sequence differences before assigning an isoform.
It can if its epitope lies in a region that isoform lacks or replaces. Isoform 3 lacks canonical residues 1–104; isoforms 2 and 3 lack 256–328 and replace 243–255. Check the antibody epitope against each isoform before interpreting a missing band.
It can if its epitope lies in a region that isoform lacks or replaces. Isoform 3 lacks canonical residues 1–104; isoforms 2 and 3 lack 256–328 and replace 243–255. Check the antibody epitope against each isoform before interpreting a missing band.
Which MSI2 modifications matter when interpreting bands?
PTM · UniProt annotates N-acetylmethionine at residue 1, phosphoserine at 6, omega-N-methylarginine at 228, and an annotation listing omega-N-methylarginine, N-acetylalanine, and phosphoserine at 261. These are UniProt canonical coordinates; antibody or paper numbering may differ. The annotations do not establish that any modification creates a resolvable band shift.
Does this guide establish induction of MSI2?
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 MSI2?
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 A02898-2 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 MSI2 bands be quantified across samples?
Quantitation · Choose the band near 35 kDa for canonical MSI2 and apply the same band selection across samples. If additional bands are included, establish that the antibody recognizes the relevant isoforms first: isoforms 2 and 3 have substantial sequence changes. Report whether the measurement represents the canonical band or combined MSI2 signal.
Why does MSI2 run near 35 kDa?
Interpretation · The canonical MSI2 sequence is 328 residues with a predicted mass of 35.2 kDa, consistent with the observed band near 35 kDa. The listed modifications alone do not establish a visible shift or explain any small difference between calculated and apparent mass.
UniProt reports that MSI2 interacts with RHOBTB2 and that this interaction is necessary for MSI2 ubiquitination and degradation. Consider differences in MSI2 abundance when comparing samples with differing RHOBTB2 activity; the feature does not predict a particular band pattern.
First compare their positions with the three annotated isoforms and check whether the antibody epitope is retained. UniProt lists modified residues and RHOBTB2-dependent ubiquitination and degradation, but these features alone cannot identify an unexpected band. No signal peptide, propeptide, or glycosylation sites are listed to support those explanations.
UniProt reports that MSI2 interacts with RHOBTB2 and that this interaction is necessary for MSI2 ubiquitination and degradation. Consider differences in MSI2 abundance when comparing samples with differing RHOBTB2 activity; the feature does not predict a particular band pattern.
First compare their positions with the three annotated isoforms and check whether the antibody epitope is retained. UniProt lists modified residues and RHOBTB2-dependent ubiquitination and degradation, but these features alone cannot identify an unexpected band. No signal peptide, propeptide, or glycosylation sites are listed to support those explanations.
Boster reagents
MSI2 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-MSI2 Antibody Picoband®
Cat # A02898-2
Real WB data
Anti-MSI2/Musashi 2 Rabbit Monoclonal Antibody
Cat # M02898
Real WB data
Anti-MSI2 Antibody
Cat # A02898
Three the supplier anti-MSI2 antibodies list human, mouse, and rat reactivity and have WB images. A02898-2 reports a band near 35 kDa across named human, rat, and mouse lysates; the other two captions describe single lysates without comparable conditions.
Which to pick: Choose A02898-2 for the most detailed WB example, including sample types and conditions. M02898 shows T47 D cell lysate, while A02898 shows EL4 cell lysate at 1 μg/mL. All three list the same species reactivity.
Source: BosterBio MSI2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96DH6.
- Human Protein Atlas. MSI2 tissue expression.
- PMC6049051 — target-verified WB comparison
- PMC11964102 — target-verified WB comparison
- PMC5253336 — target-verified WB comparison