This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
MSI2 / RNA-binding protein Musashi homolog 2 · IHC design guide
Design Immunohistochemistry for MSI2
Plan chromogenic MSI2 IHC on paraffin sections around the cytoplasmic tissue staining reported by HPA (HPA tissue IHC). This guide covers fixation consistency, antibody selection, detection and interpretation in light of the reported staining–RNA discrepancy (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MSI2 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining in tissue (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic signal in glandular, neuronal and glial cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02898-2) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 44 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | RHOBTB2 interaction enables MSI2 degradation (UniProt) | |
| Isoform / epitope | 3 isoforms; antibody epitope coverage is unknown (UniProt) |
Section 1
Recommended MSI2 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: A02898-2) with four published MSI2 IHC protocols (PMC5442418; PMC6981452; PMC5844186; PMC5665998).
| Sample | Paraffin-embedded human thyroid cancer tissue; fixative not specified (datasheet A02898-2) |
| Fixation | Image fixative and duration unreported (datasheet A02898-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A02898-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02898-2) |
| Primary antibody | Rabbit anti-MSI2, 2-5 μg/ml (datasheet A02898-2) |
| Primary incubation | Overnight at 4 °C (datasheet A02898-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02898-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MSI2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet: A02898-2). Use citrate at pH 6.0 when following either cited citrate protocol (PMC5844186; PMC5665998).
Section 2
What Is the Expected MSI2 Staining Pattern?
MSI2 is a cytoplasmic RNA-binding protein with no transmembrane segment (UniProt Q96DH6). In paraffin-section IHC, expect cytoplasmic staining in multiple cell types, including glandular, neuronal and glial cells (HPA tissue IHC). HPA rates its tissue IHC evidence Approved but reports low consistency between antibody staining and RNA expression; treat the pattern as a guide, not a universal threshold (HPA tissue IHC).
| Cytoplasmic color in adrenal, appendix or breast glandular cells. | This matches cells scored High in those tissues (HPA tissue IHC) and MSI2's cytoplasmic location (UniProt Q96DH6). Assess the named cells against adjacent structures and controls; HPA's Approved rating carries a low antibody–RNA consistency caveat (HPA tissue IHC). |
| Predominantly nuclear staining, with little cytoplasmic signal. | A nuclear-dominant pattern conflicts with the reported cytoplasmic location (UniProt Q96DH6; HPA subcellular ICC-IF). Treat it as a possible artifact: review morphology, counterstain and controls before assigning it to MSI2 (general IHC practice). |
| Strong color in an unexpected cell population. | HPA reports broad tissue expression, so cell identity alone cannot establish a false positive (HPA tissue IHC). If the pattern persists despite suitable controls, investigate antibody cross-reactivity or endogenous detection activity (general IHC practice). |
| Color spread across tissue and blank spaces without cell boundaries. | This is diffuse background rather than interpretable cytoplasmic staining (general IHC practice; UniProt Q96DH6 cytoplasm). Compare a reagent control and tissue morphology; nonspecific antibody binding or detection background may obscure a real signal (general IHC practice). |
| No signal in glandular cells of an HPA High tissue. | Adrenal, appendix and breast glandular cells are reported High, making them useful comparison sites (HPA tissue IHC). An absent result warrants a technical review, but HPA's low antibody–RNA consistency prevents treating one negative section as definitive biological absence (HPA tissue IHC). |
💡Expected MSI2 appearanceCall a positive result when identifiable glandular, neuronal or glial cells show predominantly cytoplasmic color, potentially strong in HPA High populations; nuclear-dominant or cell-free color is suspicious (HPA tissue IHC; UniProt Q96DH6; general IHC practice).
| Compartment and topology | UniProt places MSI2 in the cytoplasm, associated with polysomes, and lists no transmembrane segment (UniProt Q96DH6). Score intracellular cytoplasmic color; a membrane outline is unsupported as the expected MSI2 pattern (UniProt Q96DH6). |
| Tissue distribution and evidence strength | HPA reports High staining in selected glandular, neuronal and glial populations, and Low staining in lymph-node non-germinal-center cells (HPA tissue IHC). Its Approved rating includes low antibody–RNA consistency, so Low is not a negative control (HPA tissue IHC). |
| Antibody validation | CAB022300 is listed as IHC Approved, while HPA068990 has no supplied IHC status (HPA antibodies). Apply the catalog antibody's own IHC validation to decisions about its staining; the supplied statuses do not establish equivalent IHC performance (HPA antibodies). |
| Isoforms and epitope coverage | UniProt lists three MSI2 isoforms and two RNA-recognition motifs (UniProt Q96DH6). The supplied record gives no antibody epitope or isoform coverage, so staining cannot identify an isoform and a negative result cannot exclude every isoform (UniProt Q96DH6; HPA antibodies). |
| IF/ICC: should the compartment change? | The separate ICC-IF evidence reports mainly cytosolic MSI2 and lists both supplied antibodies as ICC Supported (HPA subcellular ICC-IF; HPA antibodies). It supports a cytosolic expectation, without establishing an IHC protocol or identical signal intensity across methods. |
| Situation | Likely cause | Next action |
|---|---|---|
| No color in an HPA High glandular population. | A technical failure is possible; HPA's antibody–RNA consistency is low (HPA tissue IHC). | Check tissue identity, morphology, the IHC-validated antibody's instructions and run controls; review retrieval and detection as general IHC checks before interpreting absence (general IHC practice). |
| A predominantly nuclear result appears. | The pattern conflicts with UniProt cytoplasm and HPA cytosol (UniProt Q96DH6; HPA subcellular ICC-IF). | Compare with the counterstain and controls, then reassess cell boundaries and nonspecific signal before scoring it as MSI2 (general IHC practice). |
| Unexpected cells stain strongly. | Broad MSI2 expression is reported, but cross-reactivity or endogenous detection activity can also color cells (HPA tissue IHC; general IHC practice). | Identify the cells morphologically and compare appropriate reagent and detection controls; report unresolved staining as uncertain (general IHC practice). |
| The whole section looks diffusely colored. | Background can obscure the cytoplasmic pattern expected for MSI2 (general IHC practice; UniProt Q96DH6). | Inspect reagent controls; review blocking, antibody concentration, washes and detection background under the laboratory's general IHC workflow (general IHC practice). |
| A lymph-node area looks faint. | HPA reports Low staining in non-germinal-center cells there (HPA tissue IHC). | Avoid using that population as a required negative; compare an HPA High population and matched controls when assessing assay performance (HPA tissue IHC; general IHC practice). |
| Two antibodies give different IHC patterns. | Only CAB022300 has a supplied IHC Approved status; HPA068990 has no supplied IHC status (HPA antibodies). | Check each antibody's IHC validation and controls separately. Do not infer that ICC Supported status predicts matching paraffin-section IHC staining (HPA antibodies; general IHC practice). |
Experiment evidence
IHC & IF images showing MSI2 staining
A02898-2 · IHCIHC analysis of MSI2 using anti-MSI2 antibody (A02898-2). MSI2 was detected in a paraffin-embedded section of human thyroid cancer tissue. H
A02898-2 · IHCIHC analysis of MSI2 using anti-MSI2 antibody (A02898-2). MSI2 was detected in a paraffin-embedded section of rat brain tissue. Heat mediate
A02898-2 · IFIF analysis of MSI2 using anti-MSI2 antibody (A02898-2). MSI2 was detected in a paraffin-embedded section of human thyroid cancer tissue. He
A02898 · IHCImmunohistochemistry of MSI2 in mouse brain tissue with MSI2 Antibody at 5 μg/mL.
A02898 · IFImmunocytochemistry of MSI2 in EL4 cells with MSI2 antibody at 5 μg/mL.
M02898 · IHCImmunohistochemical analysis of paraffin-embedded human lung carcinoma, using MSI2 Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000153944-MSI2/tissue/cerebral+cortex#imid_5818516
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000153944-MSI2/tissue/colon#imid_5818442
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000153944-MSI2/tissue/liver#imid_5818427
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000153944-MSI2/tissue/kidney#imid_5818436
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB022300; green = MSI2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB022300; green = MSI2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MSI2 IHC & IF
🧪Run caudate first and expect neuronal cells to stain (HPA: High in caudate neuronal cells). HPA detects MSI2 in all 44 scored tissues, so there is no supported negative tissue; no-primary and isotype controls carry the negative, while unstained neighboring cells, if present, show local background but are not validated MSI2-negative cells (HPA: no negative tissue rows; MSI2 detected in all 44 scored tissues).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: MSI2 is detected in all 44 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MSI2 in A-431, U-251MG, U2OS, REH, SH-SY5Y, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section, a rabbit IgG isotype control matched to the primary antibody's clonality if known, and MSI2 knockout material as a biological negative if available (caption: rabbit anti-MSI2 primary). Quench endogenous peroxidase for chromogenic detection and check caudate neuronal pigment when interpreting brown signal (caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the A02898-2 paraffin-section caption does not state a fixative (caption: fixative not stated). The caption uses heat-mediated EDTA retrieval at pH 8.0, but does not establish that retrieval is required; frozen-section performance and whether IF is easier are unreported (caption: EDTA retrieval; HPA: cytosolic ICC-IF localization). Neuronal lipofuscin in caudate can resemble brown chromogen or autofluoresce, so interpret signal against the negative controls (standard IHC/IF practice; HPA: High in caudate neuronal cells).
HPA tissue IHC evidence for MSI2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| 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 → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: MSI2 is detected in all 44 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced MSI2 IHC Tips
Troubleshoot MSI2 staining in paraffin-section chromogenic IHC using the catalog antibody’s tissue evidence and the expected cytoplasmic localisation.
How should I retrieve MSI2 in paraffin sections when staining is weak?
Use heat-mediated antigen retrieval in EDTA at pH 8.0 for MSI2 paraffin-section IHC (datasheet A02898-2). The selected thyroid cancer section was stained after this retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (A02898-2 tissue-IHC caption). If signal is weak, first check heating consistency, section adhesion, and whether the primary incubation matches that documented condition (standard IHC practice; A02898-2 tissue-IHC caption). Vary retrieval heating cautiously on paired sections, because excessive treatment can damage morphology and complicate scoring (standard IHC practice). Keep a positive control in each run and compare cytoplasmic signal under matched detection conditions (HPA subcellular; standard IHC practice).
Could fixation explain variable MSI2 staining between paraffin blocks?
MSI2-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state the fixative (A02898-2 tissue-IHC caption). Record each block’s fixative and processing history before attributing different staining to MSI2 biology (standard IHC practice). Compare blocks using the documented EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody overnight at 4°C (datasheet A02898-2; A02898-2 tissue-IHC caption). Inspect morphology and include a consistently processed positive control to identify broader processing effects (standard IHC practice). Do not infer a fixation effect from the protein’s cytoplasmic location, modifications, or HPA tissue pattern; those sources do not establish fixation sensitivity (UniProt Q96DH6; HPA tissue IHC).
What MSI2 staining pattern should I expect in a tissue section?
Score MSI2 primarily as cytoplasmic staining: UniProt places it in the cytoplasm, and HPA reports supported cytosolic localisation (UniProt Q96DH6; HPA subcellular). UniProt also associates MSI2 with polysomes and reports no transmembrane segment, so a crisp membrane-only pattern needs independent validation (UniProt Q96DH6 topology and subcellular). HPA describes ubiquitous cytoplasmic tissue expression, with high staining in hippocampal neuronal cells and cerebral cortex glial cells among its examples (HPA tissue IHC). Compare staining within the expected cell compartment against matched negative controls and adjacent well-preserved cells (HPA subcellular; standard IHC practice). Treat isolated nuclear-only staining cautiously until its specificity is supported by another method (HPA subcellular; standard IHC practice).
Can isoforms or epitope accessibility change MSI2 IHC results?
MSI2 has 3 reported isoforms, but the supplied tissue-IHC caption does not identify the antibody’s epitope or isoform coverage (UniProt Q96DH6; A02898-2 tissue-IHC caption). Its two RNA recognition motifs span residues 21–111 and 110–187, while reported modified residues include positions 6, 228, and 261 (UniProt Q96DH6). Those features justify checking antibody epitope documentation before interpreting discordant staining, but they do not establish an epitope-masking effect (UniProt Q96DH6; standard IHC practice). Compare paired sections under the documented EDTA pH 8.0 retrieval and matched detection conditions (datasheet A02898-2; standard IHC practice). If isoform specificity matters, confirm it with independently characterised reagents or orthogonal evidence (standard IHC practice).
How can I assess MSI2 by multiplex immunofluorescence?
For the separate IF/ICC workflow, look for cytosolic MSI2 and pair it with a marker identifying the cell population being examined (HPA subcellular; standard IF practice). HPA reports high tissue staining in hippocampal neuronal cells and cerebral cortex glial cells, which can guide cell-type marker selection for those tissues (HPA tissue IHC). Choose spectrally separated fluorophores and assess unstained tissue and single-colour controls, especially where tissue autofluorescence could resemble signal (standard IF practice). Because MSI2 is cytoplasmic and has no transmembrane segment, assess permeabilisation for access to a cytosolic epitope, while recognising that the antibody’s precise epitope is unspecified (UniProt Q96DH6 topology; A02898-2 tissue-IHC caption). Do not transfer the paraffin-section caption into an IF fixation claim (A02898-2 tissue-IHC caption).
How do I distinguish diffuse background from MSI2 signal in chromogenic IHC?
Expect interpretable MSI2 signal mainly in the cytoplasm, where UniProt and HPA place the protein (UniProt Q96DH6; HPA subcellular). The documented paraffin-section example used 10% goat serum, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-linked secondary for 30 minutes at 37°C (A02898-2 tissue-IHC caption). If diffuse colour persists, compare primary-omission and secondary-only controls and check whether blocking or washing changes it (standard IHC practice). Include an endogenous peroxidase block before HRP/DAB detection and inspect pigment or precipitate that may mimic chromogen (standard IHC practice). Titrate detection conditions on matched sections while retaining a positive control and assessing morphology (standard IHC practice).
How should I score MSI2 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define viable tissue regions and score cytoplasmic MSI2 in the same cell population across sections (HPA subcellular; standard IHC practice). Report the percentage of positive cells and staining intensity, or calculate an H-score from intensity grades 0–3 weighted by their cell percentages (standard IHC practice). Where cell abundance varies, report positive-cell density per mm² of analysed viable tissue and the total cells assessed (standard IHC practice). Keep retrieval, chromogen development, imaging, thresholds, and control sections consistent across comparisons (standard IHC practice). Interpret differences cautiously because HPA labels its tissue-IHC profile Approved while reporting low consistency between antibody staining and RNA expression (HPA tissue IHC).
What would make an apparent MSI2-positive result doubtful?
A convincing result shows cytoplasmic staining in preserved cells, consistent with UniProt’s cytoplasmic assignment and HPA’s supported cytosolic localisation (UniProt Q96DH6; HPA subcellular). Check whether the stained cell type fits the tissue context; HPA reports high staining in adrenal glandular cells and hippocampal neuronal cells, while listing lymph-node non-germinal center cells as low (HPA tissue IHC). Question signal confined to section edges, necrotic areas, or a membrane-only compartment, and review morphology before scoring it (UniProt Q96DH6 topology; standard IHC practice). Use primary-omission and endogenous-peroxidase controls to investigate DAB colour unrelated to the primary antibody (standard IHC practice). HPA reports low staining-to-RNA consistency, so support consequential conclusions with independent evidence (HPA tissue IHC; standard IHC practice).
Boster reagents
Best MSI2 / RNA-binding protein Musashi homolog 2 IHC Antibodies
The catalog shows anti-MSI2 IHC images from human thyroid cancer, rat brain, mouse brain, and human lung carcinoma, plus IF images from human and rat tissue and EL4 cells (catalog image captions).
Real IHC data
Anti-MSI2 Antibody ®
Cat # A02898-2
Real IHC data
Anti-MSI2 Antibody
Cat # A02898
Real IHC data
Anti-MSI2/Musashi 2 Rabbit Monoclonal Antibody
Cat # M02898
A02898-2 has paraffin-section IHC images from human thyroid cancer and rat brain, while A02898 has a mouse brain IHC image (catalog IHC captions). M02898 has a paraffin-section IHC image from human lung carcinoma (catalog IHC caption).
Which to pick: For tissue IHC, choose A02898-2 when a documented paraffin-section workflow helps: its captions specify EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody; the fixative is unreported (A02898-2 IHC captions). For IF/ICC, choose A02898: both applications are listed, with EL4-cell and mouse-brain IF images (A02898 applications; IF captions). For cross-species planning, all three list Human, Mouse, and Rat reactivity, and M02898 offers a rabbit monoclonal clone, FOB-13; the IHC images cover human and rat for A02898-2, mouse for A02898, and human for M02898 (catalog reactivity; M02898 clone; IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96DH6 (MSI2H_HUMAN, RNA-binding protein Musashi homolog 2).
- Human Protein Atlas. MSI2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MSI2 subcellular location (ICC-IF): Mainly localized to the cytosol..
- Human Protein Atlas. MSI2 antibody validation summary (2 antibodies).
- DBC2/RhoBTB2 functions as a tumor suppressor protein via Musashi-2 ubiquitination in breast cancer. Oncogene 2017 — PMC5442418.
- Immunohistochemical Analysis Revealed a Correlation between Musashi-2 and Cyclin-D1 Expression in Patients with Oral Squamous Cells Carcinoma. International journal of molecular sciences 2019 — PMC6981452.
- Musashi-2 is a prognostic marker for the survival of patients with cervical cancer. Oncology letters 2018 — PMC5844186.
- Mapping genomic and transcriptomic alterations spatially in epithelial cells adjacent to human breast carcinoma. Nature communications 2017 — PMC5665998.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:12649177 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MSI2 (RNA-binding protein Musashi homolog 2). Expected localisation Cytoplasmic staining in tissue (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02898-2). Validated IHC antibody: A02898-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 44 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human thyroid cancer tissue; fixative not specified (datasheet A02898-2); 2. Fixation: Image fixative and duration unreported (datasheet A02898-2); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A02898-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02898-2); 8. Primary antibody: Rabbit anti-MSI2, 2-5 μg/ml (datasheet A02898-2); 9. Primary incubation: Overnight at 4 °C (datasheet A02898-2). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; antibody epitope coverage is unknown (UniProt); Regulation: RHOBTB2 interaction enables MSI2 degradation (UniProt). Evidence: UniProt Q96DH6, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol., Datasheet A02898-2, PMC5442418, PMC6981452. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MSI2/immunohistochemistry-ihc-guide.