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MOB4 / MOB-like protein phocein · IHC design guide
Design Immunohistochemistry for MOB4
Plan paraffin-section MOB4 IHC around a general cytoplasmic readout (HPA tissue IHC). Compare high-staining glandular cells with adipocytes reported as not detected, and interpret staining with the cross-gene caution (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MOB4 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 | General cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in glandular and epithelial cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A11070) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+1 more · see all |
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 may include proteins from other genes (HPA tissue IHC) | |
| Regulation | Regulation not annotated (UniProt) | |
| Isoform / epitope | 3 isoforms; no processing annotated; check epitope coverage (UniProt) |
Section 1
Recommended MOB4 IHC & IF Protocols
The catalog antibody’s paraffin IHC protocol uses pH 9.0 Tris-EDTA heat retrieval (datasheet A11070). One published larval CNS protocol provides additional staining conditions (PMC6581792).
| Sample | Paraffin-embedded human Gastric adenocarcinoma tissue; fixative not specified (datasheet A11070) |
| Fixation | Image fixative and duration unreported (datasheet A11070); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A11070); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-MOB4, 1:100-1:300 (datasheet A11070) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MOB4-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteFor paraffin sections, start with pH 9.0 Tris-EDTA heat retrieval (datasheet A11070). The larval CNS protocol does not specify antigen retrieval (PMC6581792).
Section 2
What Is the Expected MOB4 Staining Pattern?
MOB4 staining should be mainly cytoplasmic, with possible perinuclear puncta and Golgi or membrane association (UniProt Q9Y3A3 localisation). High staining is reported in several glandular and epithelial cell types (HPA: tissue IHC). Interpret the pattern cautiously: HPA rates tissue staining Approved with medium RNA agreement and warns that its antibodies may detect proteins from more than one gene (HPA: reliability description). MOB4 has no annotated transmembrane segment (UniProt Q9Y3A3 topology).
| Cytoplasmic staining in colon or duodenal glandular cells, with stronger signal around some nuclei. | This fits the reported high staining in these cells (HPA: High in colon and duodenal glandular cells) and the predicted perinuclear pattern (UniProt Q9Y3A3 localisation). Assess cell type and compartment together; cytoplasmic signal alone does not resolve HPA's multiple-gene detection caveat (HPA: reliability description). |
| Strong nuclear staining dominates while the cytoplasm is faint in an otherwise positive epithelium. | A predominantly nuclear pattern conflicts with the reported general cytoplasmic tissue pattern (HPA: tissue IHC profile) and perinuclear, membrane-associated localisation (UniProt Q9Y3A3 localisation). Investigate detection background and antibody specificity before calling it MOB4; an isolated nuclear signal is insufficient to establish localisation. |
| Prominent staining appears in adipocytes or bone-marrow hematopoietic cells. | These cells are reported as not detected (HPA: adipocytes and bone-marrow hematopoietic cells). Check whether the signal follows cell outlines or nonspecific deposit. Cross-reactivity or endogenous chromogen activity are possibilities to test, rather than proven causes; HPA also cautions that its antibodies may detect proteins from more than one gene (HPA: reliability description). |
| Pale, widespread colour covers cells and surrounding tissue without a clear cytoplasmic boundary. | This is difficult to score against the reported cytoplasmic pattern (HPA: tissue IHC profile). In chromogenic IHC, incomplete blocking, excess detection reagent, or residual endogenous activity can cause diffuse background (general IHC practice). Compare a control processed without primary antibody before assigning the colour to MOB4. |
| No convincing signal appears in bronchial respiratory epithelium or esophageal squamous epithelium. | Both cell types have reported high staining (HPA: bronchus and esophagus). A blank result calls for a run-level check of section integrity, retrieval, primary-antibody application, and detection (general IHC practice). It does not establish absence of MOB4 in that specimen; HPA's reported levels are observations, not a guarantee for every section. |
💡Expected MOB4 appearanceCall a section positive when the relevant glandular or epithelial cells show clear, potentially strong cytoplasmic staining, sometimes concentrated in perinuclear puncta (HPA: High in listed glandular and epithelial cells; UniProt Q9Y3A3 localisation); dominant nuclear colour or broad staining of HPA-undetected cell types warrants investigation (HPA: tissue IHC profile and not-detected cells).
| Tissue and cell selection | HPA reports high staining across several glandular and epithelial populations, while adipocytes and bone-marrow hematopoietic cells are not detected (HPA: tissue IHC). Use a reported high-staining cell population to judge whether the run worked; the undetected populations are comparison sites, not definitive specificity controls. |
| Compartment and topology | UniProt places MOB4 in cytoplasm, perinuclear puncta, membranes, and Golgi stacks, with no transmembrane segment (UniProt Q9Y3A3 localisation and topology). Perinuclear enrichment can fit the record; membrane association does not imply a surface-only stain. Score the dominant compartment in the relevant cells. |
| Antibody interpretation | Two listed antibodies have Approved IHC status, but neither is marked IHC Enhanced in the supplied record (HPA: HPA044125 and CAB034082 antibody status). HPA reports medium agreement with RNA and cautions about detection of proteins from more than one gene (HPA: reliability description). A matching stain supports interpretation without proving MOB4-specific binding. |
| Isoforms and processing | Three isoforms are listed; the supplied record gives no isoform-specific staining or epitope map (UniProt Q9Y3A3 isoforms). It also lists no signal peptide, propeptide, or glycosylation sites (UniProt Q9Y3A3 processing). Do not infer an isoform-specific stain or a shed-product pattern from these annotations. |
| Retrieval and detection | The supplied sources give no MOB4-specific retrieval setting or fixation-sensitivity result (HPA: supplied tissue IHC; UniProt Q9Y3A3). Retrieval, blocking, and chromogen controls remain general paraffin-IHC workflow decisions (general IHC practice); their effects on MOB4 staining must be checked in the actual run. |
| Situation | Likely cause | Next action |
|---|---|---|
| A reported high-staining epithelium is blank. | A failed retrieval, omitted primary antibody, or detection problem can produce a blank IHC run (general IHC practice); the expected high cell-level staining comes from HPA (HPA: bronchus or esophagus). | Check a reported positive cell population on the same run, then verify reagent application, retrieval record, and chromogen development (general IHC practice). Do not score a blank section as a biological negative until the run control works. |
| Colour concentrates in nuclei instead of cytoplasm. | Dominant nuclear staining conflicts with the reported cytoplasmic tissue pattern (HPA: tissue IHC profile) and UniProt's perinuclear and Golgi localisation (UniProt Q9Y3A3 localisation). | Inspect a no-primary control and compare cell boundaries with the counterstain (general IHC practice). Reassess detection background and antibody specificity before accepting a nuclear localisation. |
| Adipocytes or bone-marrow hematopoietic cells stain strongly. | Both are listed as not detected (HPA: tissue IHC). Endogenous chromogen activity or antibody cross-reactivity are testable explanations (general IHC practice; HPA: multiple-gene detection caution). | Compare no-primary and detection controls, and examine whether a reported high-staining epithelial population shows the expected cytoplasm (general IHC practice; HPA: tissue IHC profile). Treat persistent staining in these cells cautiously. |
| The entire section has weak brown haze. | Diffuse colour lacks the reported cell-associated cytoplasmic pattern (HPA: tissue IHC profile). Inadequate blocking, excessive detection signal, or residual endogenous activity can contribute (general IHC practice). | Compare the no-primary control, inspect wash and blocking steps, and adjust detection conditions within the assay workflow (general IHC practice). Score only cell-associated signal above local background. |
| A second antibody gives a different cell pattern. | The two listed antibodies are IHC Approved, while HPA reports medium RNA agreement and possible detection from more than one gene (HPA: antibody status and reliability description). Agreement is therefore useful evidence, and disagreement needs investigation. | Compare the same tissue compartments and cell types under matched scoring criteria; review each antibody's validation and controls before choosing a pattern (general IHC practice). Do not describe either reagent as IHC Enhanced from this record (HPA: supplied antibody status). |
| IF/ICC Q&A: Do perinuclear fluorescent puncta validate MOB4 localisation? | Perinuclear puncta are consistent with UniProt's localisation annotation, but the supplied HPA subcellular record has no main location or ICC-IF images (UniProt Q9Y3A3 localisation; HPA: subcellular record). | Treat puncta as a localisation hypothesis, not an independently validated IF/ICC result. Use the separate IF/ICC guide for assay design; this IHC section supplies no IF/ICC protocol (HPA: subcellular record). |
Experiment evidence
IHC & IF images showing MOB4 staining
A11070 · IHCImmunohistochemical analysis of paraffin-embedded human Gastric adenocarcinoma. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDT
A19781 · IHCImmunohistochemical analysis of paraffin-embedded human Gastric adenocarcinoma. 1, Antibody was diluted at 1:200(4° overnight). 2, Tris-EDTA
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000115540-MOB4/tissue/cerebral+cortex#imid_13920448
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000115540-MOB4/tissue/colon#imid_13920382
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000115540-MOB4/tissue/liver#imid_13920375
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000115540-MOB4/tissue/kidney#imid_13920392
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MOB4 IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the colon slide, assess non-glandular cells for background staining, but do not assume they are MOB4-negative without validation.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for MOB4; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and host-species- and clonality-matched isotype controls, plus a biological specificity control using MOB4 knockout material or peptide blocking where applicable (standard IHC practice). For colon, quench endogenous peroxidase and check for endogenous biotin if using avidin–biotin detection (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A11070 paraffin-section caption does not state a fixative (caption: fixative unreported). Tris-EDTA at pH 9.0 is a retrieval starting point, not an established requirement (caption: Tris-EDTA pH 9.0). The supplied evidence does not establish whether frozen sections or IF are easier; assess colonic mucus and endogenous peroxidase as possible chromogenic background (standard IHC practice).
HPA tissue IHC evidence for MOB4
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). 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 → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced MOB4 IHC Tips
Troubleshoot MOB4 staining in paraffin sections by checking retrieval, compartmental pattern, controls, and scoring before drawing biological conclusions.
Which retrieval conditions should I start with for MOB4 in paraffin sections?
Start with heat-mediated antigen retrieval in Tris-EDTA at pH 9.0 (datasheet A11070). That condition accompanies MOB4 staining in paraffin-embedded human gastric adenocarcinoma with antibody A11070 at 1:200 overnight at 4°C (datasheet A11070). If staining is weak, compare a shorter and longer heating cycle on adjacent sections while holding antibody concentration, detection, and development constant (general IHC practice). Include a section processed without primary antibody to reveal detection background, and compare signal within equivalent tissue regions (general IHC practice). The caption does not report the fixative, so the documented retrieval condition cannot establish how a particular fixation method affects staining (datasheet A11070).
How should I assess whether fixation is limiting MOB4 staining?
Target-specific MOB4 sensitivity to fixation is unknown from the supplied evidence; the paraffin-section caption does not state a fixative (datasheet A11070). Record the fixative and fixation duration for each specimen, then compare adjacent sections processed with the same Tris-EDTA pH 9.0 retrieval and detection conditions (general IHC practice; datasheet A11070). Use the documented 1:200 primary incubation overnight at 4°C as a reproducible starting condition for A11070 (datasheet A11070). Assess preserved tissue structure and compare expected cytoplasmic staining in comparable viable cells before attributing a difference to fixation (HPA: general cytoplasmic expression; general IHC practice). Neither the HPA staining profile nor MOB4 topology establishes a fixation-specific effect (HPA: tissue IHC; UniProt Q9Y3A3 topology).
Where should convincing MOB4 chromogenic staining appear within cells?
Look primarily for cytoplasmic staining, with a possible perinuclear punctate component associated with Golgi stacks and membranes (HPA: general cytoplasmic expression; UniProt Q9Y3A3 subcellular location). MOB4 has no annotated transmembrane segment, so a sharply outlined cell surface pattern alone does not match its recorded topology (UniProt Q9Y3A3 topology). Compare staining at high magnification in viable cells and use the counterstain to distinguish perinuclear signal from nuclear staining (general IHC practice). Keep the interpretation at the cellular-compartment level: chromogenic signal cannot by itself prove Golgi association or STRIPAK membership (UniProt Q9Y3A3 subcellular location and subunit; general IHC practice). Check a no-primary section when diffuse color obscures the proposed compartment (general IHC practice).
Can this IHC result distinguish MOB4 isoforms or identify the bound epitope?
MOB4 has 3 recorded isoforms, but the supplied IHC caption does not specify an epitope or establish isoform selectivity for A11070 (UniProt Q9Y3A3 isoforms; datasheet A11070). Treat a positive chromogenic signal as antibody reactivity in tissue rather than as proof of one splice form (general IHC interpretation). MOB4 has no annotated glycosylation sites or modified residues in the supplied record, which does not identify the antibody-binding site (UniProt Q9Y3A3 processing and modifications). If isoform identity matters, obtain epitope mapping and isoform-specific validation before assigning staining to isoform 1, 2, or 3 (UniProt Q9Y3A3 isoforms; general antibody-validation practice). Compare any validation material under the same retrieval and detection conditions used for the tissue sections (general IHC practice).
How can IF help assess the MOB4 pattern seen by chromogenic IHC?
Use IF as a separate localisation check alongside the paraffin-section IHC result; the supplied A11070 caption documents tissue IHC, not an IF protocol (datasheet A11070). Multiplex MOB4 with a marker for the expected cell population in the specimen, then judge whether the signals occupy the same cells without assuming that overlap proves a molecular interaction (general IF practice). Choose a fluorophore channel after examining an unstained section for tissue autofluorescence, and include single-label controls to assess bleed-through (general IF practice). MOB4 is cytoplasmic and associated with Golgi-stack membranes without an annotated transmembrane segment, so optimise permeabilisation to expose intracellular epitopes while preserving the perinuclear pattern (UniProt Q9Y3A3 subcellular location and topology; general IF practice). Do not transfer the IHC incubation or retrieval conditions to IF without validation (datasheet A11070; general IF practice).
What should I check when MOB4 staining is diffuse or unusually strong?
Compare the stained section with a no-primary control and inspect tissue edges, damaged areas, and deposits of chromogen before calling diffuse color MOB4 positive (general IHC practice). Check the peroxidase block and shorten chromogen development if background appears in the control; these are general chromogenic IHC steps, not MOB4-specific findings (general IHC practice). For A11070, the documented primary dilution is 1:200 overnight at 4°C, followed by secondary antibody at 1:200 for 45 minutes at room temperature (datasheet A11070). If background persists, compare a primary-antibody dilution series while keeping retrieval, section thickness, and development matched (general IHC practice). Retain only signal whose cellular distribution remains interpretable against the control (general IHC practice).
How should I score MOB4 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and regions of interest before scoring, and exclude necrosis, folds, and tissue edges using the same rules across sections (general IHC practice). For cellular staining, record the percentage of positive cells and intensity from 0–3; an H-score sums each intensity multiplied by its percentage of cells, yielding 0–300 (general IHC practice). Normalise comparisons to the number of eligible cells or evaluable tissue area, and report the denominator with each result (general IHC practice). Score cytoplasmic signal separately from any perinuclear puncta because both patterns are relevant to MOB4 localisation (HPA: general cytoplasmic expression; UniProt Q9Y3A3 subcellular location). Keep retrieval, staining batch, and image analysis thresholds consistent for compared sections (general IHC practice).
How can I separate plausible MOB4 staining from artefact?
A plausible positive has interpretable cytoplasmic staining, potentially with perinuclear puncta, in viable cells and exceeds the matched no-primary background (HPA: general cytoplasmic expression; UniProt Q9Y3A3 subcellular location; general IHC practice). Strong staining confined to nuclei, section edges, or necrotic regions calls for review of compartment assignment and tissue artefact (UniProt Q9Y3A3 subcellular location; general IHC practice). Check endogenous peroxidase signal with the no-primary control before assigning chromogen to MOB4 (general IHC practice). HPA reports high staining in glandular cells of the colon and no detection in adipocytes of adipose tissue, but its tissue IHC entry warns that the antibody targets protein from more than one gene (HPA: colon, adipose tissue, and reliability description). Require an independent specificity check before interpreting a biological difference as MOB4-specific (HPA: reliability description; general antibody-validation practice).
Boster reagents
Best MOB4 / MOB-like protein phocein IHC Antibodies
Two anti-MOB4 antibodies have real IHC images from human gastric adenocarcinoma paraffin sections (catalog IHC captions). Both list IF and human, mouse, and rat reactivity (catalog applications and reactivity).
Real IHC data
Anti-Phocein MOB4 Antibody
Cat # A11070
Real IHC data
Anti-MOBL3 Antibody
Cat # A19781
A11070 will render with an IHC image from human gastric adenocarcinoma paraffin sections; IF is listed, and human, mouse, and rat reactivity is reported (A11070 catalog IHC caption, applications, and reactivity). A19781 will render with its own IHC image from human gastric adenocarcinoma paraffin sections; IF and ICC are listed, and human, mouse, and rat reactivity is reported (A19781 catalog IHC caption, applications, and reactivity).
Which to pick: For tissue IHC, either SKU has a paraffin-section image using primary antibody at 1:200 after Tris-EDTA retrieval at pH 9.0 (A11070 and A19781 IHC captions); neither caption reports the fixative. For IF/ICC, choose A19781 because both applications are listed and it is described as polyclonal (A19781 catalog applications and dilution_raw); A11070 lists IF, but neither SKU has an IF image in the payload (catalog applications and image alts). For mouse or rat samples, both list reactivity with those species, while each supplied IHC image shows only human tissue (catalog reactivity and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9Y3A3 (PHOCN_HUMAN, MOB-like protein phocein).
- Human Protein Atlas. MOB4 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MOB4 subcellular location (ICC-IF): Highest expression in SuSa: 82.1 nTPM.
- Human Protein Atlas. MOB4 antibody validation summary (2 antibodies).
- Mob4-dependent STRIPAK involves the chaperonin TRiC to coordinate myofibril and microtubule network growth. PLoS genetics 2022 — PMC9258817.
- The STRIPAK complex is required for radial sorting and laminin receptor expression in Schwann cells. Cell reports 2025 — PMC12035956.
- STRIPAK Members Orchestrate Hippo and Insulin Receptor Signaling to Promote Neural Stem Cell Reactivation. Cell reports 2019 — PMC6581792.
- PubMed PMID:10810093 — UniProt-cited evidence.
- PubMed PMID:11230166 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MOB4 (MOB-like protein phocein). Expected localisation General cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A11070). Validated IHC antibody: A11070 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human Gastric adenocarcinoma tissue; fixative not specified (datasheet A11070); 2. Fixation: Image fixative and duration unreported (datasheet A11070); 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: Tris-EDTA pH 9.0 (datasheet A11070); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-MOB4, 1:100-1:300 (datasheet A11070); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining may include proteins from other genes (HPA tissue IHC); Isoform / epitope: 3 isoforms; no processing annotated; check epitope coverage (UniProt); Regulation: Regulation not annotated (UniProt). Evidence: UniProt Q9Y3A3, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in SuSa: 82.1 nTPM, Datasheet A11070, PMC6581792. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MOB4/immunohistochemistry-ihc-guide.