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MOSPD1 / Motile sperm domain-containing protein 1 · IHC design guide
Design Immunohistochemistry for MOSPD1
Plan MOSPD1 IHC-P around observed cytoplasmic staining (HPA tissue IHC) and annotated ER/Golgi membranes (UniProt). Compare high staining in Purkinje cells with undetected staining in adipocytes (HPA tissue IHC), and validate the catalog antibody in human sections because its reported IHC test used mouse tissue (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MOSPD1 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 (HPA tissue IHC); ER/Golgi membranes (UniProt) | |
| Staining pattern | High cytoplasmic staining in Purkinje cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Cerebellum+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 paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A15298) | |
| Caveat | Human IHC is untested for the catalog antibody (datasheet) | |
| Regulation | No expression regulator specified (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage and membrane side unknown (UniProt) |
Section 1
Recommended MOSPD1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by one published MOSPD1 IHC protocol for breast tumor tissue (PMC11453086).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A15298); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| 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-MOSPD1, 1:100–1:500 starting range (standard) |
| 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 | MOSPD1-positive staining in purkinje cells of cerebellum (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen retrieval); the published method used boiling citrate for 15–20 min (PMC11453086).
Section 2
What Is the Expected MOSPD1 Staining Pattern?
In paraffin IHC, expect mainly cytoplasmic MOSPD1 staining, strongest in cerebellar Purkinje cells and moderate in several other listed cell types (HPA tissue IHC: general cytoplasmic expression; Purkinje cells High). MOSPD1 is annotated at endoplasmic reticulum and Golgi membranes, with two transmembrane segments (UniProt Q9UJG1). Treat the tissue pattern as a guide: HPA rates its IHC evidence Approved, with medium agreement between staining and RNA data and external verification pending (HPA tissue IHC).
| Purkinje cell cytoplasm stains strongly; surrounding tissue is less prominent. | This matches the clearest listed positive cell type (HPA tissue IHC: cerebellar Purkinje cells High). Assess signal within identified cells, since HPA reports a general cytoplasmic IHC pattern rather than a distinctive structure visible in every paraffin section (HPA tissue IHC). |
| Glandular, basal, glial or squamous epithelial cells show moderate cytoplasmic staining. | Moderate signal fits the listed cells in adrenal gland, appendix, breast, bronchus, caudate, cerebral cortex and cervix (HPA tissue IHC: Medium). Compare like cell types within a section; the tissue name alone does not specify that every cell should stain (HPA tissue IHC). |
| A putative IHC positive is confined to nuclei, with no convincing cytoplasmic signal. | Recheck compartment assignment and controls: nuclear-only chromogen differs from the general cytoplasmic tissue IHC profile (HPA tissue IHC). Do not automatically dismiss nuclear signal across methods, because approved ICC-IF images place MOSPD1 in the nucleoplasm and vesicles (HPA subcellular ICC-IF). |
| Strong colour appears in an unexpected cell type, or spreads evenly beyond cell boundaries. | Check for cross-reactivity, endogenous detection activity or background before scoring it as MOSPD1 (general IHC practice). HPA reports adipocytes and ovarian stroma cells as Not detected, while several other cells have Low signal; those observations are useful comparisons, not absolute specificity tests (HPA tissue IHC). |
| A known positive compartment has no detectable chromogen. | Absent Purkinje cell staining conflicts with the strongest listed IHC observation (HPA tissue IHC: Purkinje cells High). First evaluate staining controls and tissue preservation (general IHC practice); HPA's Approved rating has medium RNA agreement and awaits external verification, so a single negative section is not proof of absent protein (HPA tissue IHC). |
💡Expected MOSPD1 appearanceCall a result positive when identifiable Purkinje cells have strong cytoplasmic chromogen, or listed moderate-positive cell types have cell-associated cytoplasmic chromogen (HPA tissue IHC); isolated nuclear-only colour or widespread colour outside cells calls for control review before scoring (HPA tissue IHC; general IHC practice).
| Compartment evidence | UniProt places MOSPD1 at endoplasmic reticulum and Golgi membranes and records transmembrane segments at residues 159–179 and 191–211 (UniProt Q9UJG1). HPA tissue IHC describes general cytoplasmic expression; it does not resolve those organelles in a routine chromogenic section (HPA tissue IHC; general IHC practice). |
| Cell type and signal range | HPA lists Purkinje cells as High, seven specified cell populations as Medium, and other populations as Low or Not detected (HPA tissue IHC). Score the cell population examined rather than assigning one intensity to an entire tissue (general IHC practice). |
| Antibody evidence | The HPA antibody HPA031158 is Approved for IHC and ICC, while the tissue IHC summary reports medium staining-to-RNA consistency and pending external verification (HPA antibodies; HPA tissue IHC). An Approved result supports pattern comparison but does not independently establish every signal's specificity (HPA tissue IHC). |
| Isoforms and epitope | UniProt lists three MOSPD1 isoforms and an MSP domain at residues 16–143 (UniProt Q9UJG1). The supplied evidence gives no antibody epitope, so it cannot establish which isoforms the IHC antibody detects or predict an isoform-specific staining pattern. |
| Processing and modification | The supplied UniProt record lists a 1–213 chain, no signal peptide or propeptide, and no annotated glycosylation or modified residues (UniProt Q9UJG1). These annotations provide no basis to invoke cleavage, shedding or a modification-dependent IHC pattern; they also do not establish fixation sensitivity. |
| IF/ICC question: should its pattern match IHC exactly? | No exact match is established: HPA reports approved nucleoplasm and vesicle localisation in ICC-IF images from A-431, U-251MG and U2OS, while tissue IHC is generally cytoplasmic (HPA subcellular ICC-IF; HPA tissue IHC). Interpret each application with its own controls and readout (general IHC/IF practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Purkinje cells are unstained. | The expected strong reference pattern is missing (HPA tissue IHC: Purkinje cells High); an IHC workflow or sample problem is possible (general IHC practice). | Verify tissue identity, positive and negative controls, primary antibody use and detection reagents; review antigen retrieval against the IHC-validated antibody's instructions (general IHC practice). No MOSPD1-specific retrieval or fixation response is supplied. |
| Every cell shows similar weak colour. | Diffuse colour can reflect nonspecific background or detection activity (general IHC practice), and obscures HPA's cell-specific intensity differences (HPA tissue IHC). | Inspect the no-primary control, blocking, washing and chromogen development; adjust these general IHC steps before comparing cell populations (general IHC practice). |
| Colour appears mainly outside cells or along section edges. | The distribution does not resemble HPA's general cytoplasmic IHC profile (HPA tissue IHC); staining artefact is possible (general IHC practice). | Inspect section quality and control slides, then repeat with consistent washing and detection conditions if needed (general IHC practice). |
| Adipocytes or ovarian stroma cells stain strongly. | That contrasts with Not detected observations for those specific cell populations (HPA tissue IHC); cross-reactivity or endogenous activity is possible (general IHC practice). | Confirm cell identity and examine no-primary and detection controls. Compare the same run with a listed positive population before assigning the colour to MOSPD1 (HPA tissue IHC; general IHC practice). |
| Chromogenic signal seems exclusively nuclear. | Nuclear-only staining differs from HPA's general cytoplasmic tissue IHC profile, although nucleoplasm is an approved ICC-IF location (HPA tissue IHC; HPA subcellular ICC-IF). | Check morphology, counterstain and control slides, then score IHC and IF/ICC within their respective evidence sets; do not transfer an ICC-IF localisation call directly to paraffin IHC (general IHC/IF practice). |
| A listed moderate-positive population stains inconsistently. | HPA reports Medium staining in specified cell types and only medium agreement between IHC staining and RNA expression (HPA tissue IHC). Section-to-section variation or workflow variation may also affect an IHC comparison (general IHC practice). | Compare identified cells in matched sections and review staining controls across the run. Record the observed intensity rather than forcing a Medium score; seek independent confirmation if the conclusion depends on a disputed pattern (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing MOSPD1 staining
A15298 · IHCImmunohistochemistry of MOSPD1 in mouse testis tissue with MOSPD1 antibody at 2.5 μg/ml.
A15298 · IFImmunofluorescence of MOSPD1 in mouse testis tissue with MOSPD1 antibody at 20 μg/ml.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000101928-MOSPD1/tissue/cerebral+cortex#imid_7933860
HPA · CerebellumHuman Protein Atlas IHC, Cerebellum. Source: https://www.proteinatlas.org/ENSG00000101928-MOSPD1/tissue/cerebellum#imid_7933863
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000101928-MOSPD1/tissue/colon#imid_7930410
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000101928-MOSPD1/tissue/liver#imid_7930419
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA031158; green = MOSPD1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA031158; green = MOSPD1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MOSPD1 IHC & IF
🧪Run cerebellum first and expect Purkinje cells to stain (HPA: High in cerebellar Purkinje cells). Use adipose tissue as the negative tissue, focusing on adipocytes (HPA: Not detected in adipocytes); on the cerebellar slide, use neighboring cells without specific signal as an internal background reference without assigning them a MOSPD1-negative identity (HPA: cerebellar Purkinje cells High).
Positive control tissue: Cerebellum (Purkinje cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MOSPD1 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved), Vesicles (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, an isotype control matched to the primary antibody’s host species and immunoglobulin class where applicable, and a MOSPD1-knockout biological negative if available (standard IHC practice). For chromogenic cerebellar IHC, control endogenous peroxidase; for IF, check an unstained section for neuronal autofluorescence (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependence are unreported in the supplied evidence; the selected A15298 mouse-testis IHC caption also leaves the fixative unreported (A15298 tissue-IHC caption). The supplied evidence does not establish whether frozen sections or IF are easier than paraffin IHC (HPA: ICC-IF images in A-431, U-251MG and U2OS; A15298 tissue-IHC caption). In cerebellar IF, neuronal autofluorescence can complicate signal assessment (standard IF practice).
HPA tissue IHC evidence for MOSPD1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Basal cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced MOSPD1 IHC Tips
Troubleshoot MOSPD1 staining in paraffin sections by checking retrieval, compartment, controls, and scoring before interpreting signal.
What retrieval conditions should I start with for MOSPD1 in paraffin sections?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval setting). Let sections cool in buffer, then rinse consistently before blocking so comparisons reflect the same treatment (standard IHC practice). MOSPD1 has an MSP domain at residues 16–143 and transmembrane segments at 159–179 and 191–211, but the antibody epitope is unspecified, so topology cannot establish whether retrieval exposed it (UniProt Q9UJG1 topology; antibody epitope not supplied). If staining remains weak, compare a limited alternative retrieval condition on adjacent sections while keeping detection and exposure to chromogen constant (standard IHC practice).
How should I investigate weak staining after fixation?
MOSPD1-specific sensitivity to fixation is unknown; the supplied mouse testis caption gives an antibody concentration of 2.5 µg/ml but does not state the fixative (selected A15298 tissue-IHC caption). Record fixative, fixation duration, tissue thickness, and processing history for each paraffin block before comparing staining (standard IHC practice). Run sections from a consistently processed control block alongside the test material and use the same retrieval and detection conditions (standard IHC practice). If signal varies by processing batch, compare matched material processed under documented conditions; neither HPA staining patterns nor MOSPD1 topology establishes a target-specific fixation effect (HPA tissue IHC; UniProt Q9UJG1 topology).
Which compartments should count as plausible MOSPD1 staining?
For chromogenic IHC, assess cytoplasmic staining in its cellular context: HPA describes a general cytoplasmic tissue pattern, while UniProt places MOSPD1 at endoplasmic reticulum and Golgi membranes (HPA tissue IHC; UniProt Q9UJG1 subcellular location). HPA immunofluorescence also reports approved nucleoplasmic and vesicular localisation, so a nuclear component merits checking rather than automatic exclusion (HPA subcellular). Compare morphology and signal across adjacent cells and serial sections, including a no-primary control for diffuse chromogen (standard IHC practice). Document cytoplasmic, nuclear, and punctate components separately; a chromogenic pattern alone cannot establish organelle identity or resolve differences between the reported locations (standard IHC interpretation; HPA tissue IHC; HPA subcellular).
Could isoforms or epitope placement explain inconsistent staining?
MOSPD1 has 3 reported isoforms, an MSP domain at residues 16–143, and 2 transmembrane segments near its C terminus (UniProt Q9UJG1). The supplied antibody information does not identify its epitope or establish which isoforms it detects (supplied antibody evidence). Check the antibody's epitope documentation before interpreting a negative paraffin section as absence of every isoform (standard IHC interpretation; UniProt Q9UJG1 isoforms). Compare staining with a second independently validated epitope or an orthogonal expression measure when isoform coverage matters (standard IHC validation practice). Record the retrieval condition and control tissue used for each antibody because accessibility can change after processing (standard IHC practice).
How can IF help assess an ambiguous MOSPD1 IHC pattern?
Use IF as a separate localisation check, since HPA reports approved nucleoplasmic and vesicular MOSPD1 signal in its subcellular dataset (HPA subcellular). Multiplex with a validated marker for the cell type under study, then assess whether MOSPD1 and that marker occur in the same cells (standard IF practice). Choose spectrally separated fluorophores after examining tissue autofluorescence in unstained controls, especially when judging weak puncta (standard IF practice). MOSPD1 contains 2 transmembrane segments, but the antibody epitope's membrane side is unspecified; optimise permeabilisation against that side once established, using matched controls (UniProt Q9UJG1 topology; standard IF practice). Do not transfer the mouse testis IHC caption's preparation assumptions to IF (selected A15298 tissue-IHC caption).
How do I distinguish MOSPD1 signal from chromogenic background?
Inspect a no-primary section, an isotype control where appropriate, and matched positive and lower-expression regions before assigning weak brown deposits to MOSPD1 (standard IHC practice). Block endogenous peroxidase for peroxidase-based detection and compare a brief DAB development series under identical illumination and counterstain conditions (standard chromogenic IHC practice). HPA reports high staining in cerebellar Purkinje cells and no detection in adipocytes, which can inform control selection when suitable material is available (HPA tissue IHC). Edge accentuation, necrotic deposits, or signal reproduced without primary antibody argues for artefact rather than cell-associated antigen (standard IHC interpretation). Recheck antibody concentration and wash conditions before increasing retrieval intensity (standard IHC practice).
What scoring scheme makes MOSPD1 IHC results comparable? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring because tissue IHC shows a general cytoplasmic pattern, whereas HPA IF also reports nucleoplasmic and vesicular signal (HPA tissue IHC; HPA subcellular). For cytoplasmic chromogen, report percentage of positive eligible cells and an H-score using intensity categories 0–3, or measure positive area per mm² when cell boundaries are unreliable (standard IHC quantification practice). Normalise counts to the number of eligible, intact cells and area measurements to evaluable tissue area, excluding folds and necrosis (standard IHC quantification practice). Keep threshold, counterstain, image settings, and region selection fixed across groups, then report compartments separately (standard IHC quantification practice).
When should an apparent MOSPD1-positive section be questioned?
A convincing result shows reproducible cell-associated staining with a plausible compartment and appropriate control behaviour (standard IHC interpretation). Cytoplasmic staining is consistent with the HPA tissue profile and ER/Golgi membrane annotation; nucleoplasmic signal has separate HPA IF support but should be assessed independently in chromogenic sections (HPA tissue IHC; UniProt Q9UJG1 subcellular location; HPA subcellular). Question signal confined to section edges, necrosis, or cells staining in the no-primary control, and evaluate endogenous peroxidase when using peroxidase detection (standard IHC interpretation). HPA lists high Purkinje-cell staining and undetected adipocyte staining, but its tissue-IHC reliability is approved with medium RNA consistency and pending external verification (HPA tissue IHC).
Boster reagents
Best MOSPD1 / Motile sperm domain-containing protein 1 IHC Antibodies
A15298 has mouse testis IHC and IF images (catalog image captions). Human, mouse, and rat reactivity is listed, but IHC and IF validation is reported only in mouse samples (catalog applications and validation notes).
Real IHC data
Anti-MOSPD1 Antibody
Cat # A15298
A15298 will render with an IHC image of mouse testis stained at 2.5 μg/ml (A15298 IHC image caption). Its IF image shows mouse testis stained at 20 μg/ml (A15298 IF image caption).
Which to pick: Choose A15298 for tissue IHC: IHC-P is listed and mouse testis IHC is shown, but the image caption does not report the section processing or fixative (A15298 applications; A15298 IHC image caption). A15298 also has mouse testis IF data; ICC validation is unreported (A15298 IF image caption; catalog validation notes). For human or rat samples, A15298 lists reactivity in those species, but its IHC and IF validation is reported only in mouse samples (catalog reactivity and validation notes).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UJG1 (MSPD1_HUMAN, Motile sperm domain-containing protein 1).
- Human Protein Atlas. MOSPD1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MOSPD1 subcellular location (ICC-IF): Localized to the nucleoplasm and vesicles..
- Human Protein Atlas. MOSPD1 antibody validation summary (1 antibodies).
- Deciphering MOSPD1's impact on breast cancer progression and therapeutic response. Biology direct 2024 — PMC11453086.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15772651 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MOSPD1 (Motile sperm domain-containing protein 1). Expected localisation Cytoplasmic staining (HPA tissue IHC); ER/Golgi membranes (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A15298 at 1:100–1:500 (standard starting range) (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A15298); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 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-MOSPD1, 1:100–1:500 starting range (standard); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A15298); Caveat: Human IHC is untested for the catalog antibody (datasheet); Isoform / epitope: 3 isoforms; epitope coverage and membrane side unknown (UniProt); Regulation: No expression regulator specified (UniProt). Evidence: UniProt Q9UJG1, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm and vesicles., Datasheet A15298, PMC11453086. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MOSPD1/immunohistochemistry-ihc-guide.