MOSPD2 / Motile sperm domain-containing protein 2 · IHC design guide

Design Immunohistochemistry for MOSPD2

Plan chromogenic MOSPD2 IHC in paraffin sections using the catalog antibody's 2–5 μg/ml range (datasheet A16695-1). Assess cytoplasmic staining while accounting for the uncertain tissue evidence (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for MOSPD2 (IHC for MOSPD2): expected localisation Cytoplasmic staining (HPA tissue IHC); ER membrane (UniProt), antibody A16695-1, validated IHC image, and IHC protocol steps — Printable **MOSPD2** IHC protocol sheet — expected localisation Cytoplasmic staining (HPA tissue IHC); ER membrane (UniProt), antibody A16695-1, controls and protocol steps. Open the full MOSPD2 IHC guide →

Navigate to: Protocols Staining pattern Controls Antibodies

MOSPD2 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 membrane (UniProt)
	Staining pattern	Cytoplasmic staining in most tissues (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet A16695-1)
	Positive control ⓘ	Appendix+4 more · see all
	Negative control ⓘ	Adipose tissue+4 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep fixation consistent across paraffin sections (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 A16695-1)
	Caveat	Staining has low concordance with RNA expression (HPA tissue IHC)
	Regulation	CD14(+) monocytes enriched; neutrophils lower (UniProt)
	Isoform / epitope	2 isoforms; cytoplasmic 1–496, TM 497–518; epitope map unknown (UniProt)

Section 1

Recommended MOSPD2 IHC & IF Protocols

The catalog antibody uses EDTA pH 8.0 retrieval (datasheet A16695-1). The published paraffin-section protocol below uses citrate retrieval (PMC9338958).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human brain tissue; fixative not specified (datasheet A16695-1)
Fixation	Image fixative and duration unreported (datasheet A16695-1); 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 A16695-1); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A16695-1)
Primary antibody	Rabbit anti-MOSPD2, 2-5 μg/ml (datasheet A16695-1)
Primary incubation	Overnight at 4 °C (datasheet A16695-1)
Detection	HRP-conjugated secondary, DAB chromogen (datasheet A16695-1)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	MOSPD2-positive staining in glandular cells of appendix (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control.

💡Decision noteTry EDTA pH 8.0 first for the catalog antibody (datasheet A16695-1); citrate retrieval at 125 °C is a published alternative (PMC9338958).

Section 2

What Is the Expected MOSPD2 Staining Pattern?

MOSPD2 is an endoplasmic reticulum (ER) membrane protein with a cytoplasmic region at residues 1–496 and a transmembrane segment at 497–518 (UniProt Q8NHP6 topology). In paraffin IHC, expect cytoplasmic staining in reported positive cells, including appendix glandular cells and bone marrow hematopoietic cells (HPA: Medium). CD14(+) monocytes express MOSPD2 highly (UniProt Q8NHP6 tissue specificity). Treat the tissue pattern as provisional because HPA rates its IHC profile Uncertain (HPA: Uncertain).

What am I looking at on my slide?
Cytoplasmic stain in appendix glandular cells or bone marrow hematopoietic cells.	This fits the reported Medium staining in those cell populations (HPA: tissue IHC). Assess the named cells within the section; a positive result in one population does not imply that every cell in the tissue should stain. HPA reports low consistency between antibody staining and RNA expression, so morphology alone cannot establish specificity (HPA: Uncertain).
Predominantly nuclear staining, with little cytoplasmic signal.	A predominantly nuclear pattern conflicts with the expected ER membrane location and cytoplasmic topology (UniProt Q8NHP6 subcellular location and topology). Consider nonspecific staining or a detection artefact. Review the no-primary control and whether the signal follows nuclei or other tissue structures before scoring the section as MOSPD2 positive.
Strong staining in a cell population reported as undetected.	For example, HPA reports Not detected in adipocytes and cardiomyocytes (HPA: tissue IHC). Strong signal in those particular cells raises concern about cross-reactivity or endogenous detection activity; it does not mean the entire tissue must be blank. Compare cell morphology and controls, while remembering that HPA's tissue IHC assessment is Uncertain (HPA: Uncertain).
Diffuse colour across cells and tissue spaces, without a clear cellular pattern.	Widespread, poorly localized signal is difficult to reconcile with an ER-associated cytoplasmic result (UniProt Q8NHP6 subcellular location; HPA: cytoplasmic tissue profile). Background from the detection system or insufficient blocking is plausible in routine chromogenic IHC. Compare a no-primary control and avoid scoring diffuse haze as target staining.
No staining in appendix glandular cells or bone marrow hematopoietic cells.	Those cells have reported Medium staining and are useful comparison populations (HPA: tissue IHC), but HPA does not establish them as definitive positive controls because its tissue IHC profile is Uncertain (HPA: Uncertain). Check tissue identity, section quality, retrieval, antibody dilution and detection controls before interpreting an absent signal as biological absence.

💡Expected MOSPD2 appearanceA plausible positive result is distinct cytoplasmic staining of the relevant cells, potentially around Medium intensity in appendix glandular or bone marrow hematopoietic cells (HPA: tissue IHC), whereas dominant nuclear colour, staining confined to reported undetected cells, or diffuse tissue-wide haze should prompt control checks (UniProt Q8NHP6 ER location; HPA: Uncertain).

How each factor affects the staining
Cell population within the tissue	HPA's levels apply to specified cells: colon glandular cells are Medium, while glomerular cells in kidney are Not detected (HPA: tissue IHC). Identify and score the named population rather than assigning its level to the whole organ.
ER topology	MOSPD2 has one C-terminal transmembrane segment at residues 497–518; residues 1–496 face the cytoplasm (UniProt Q8NHP6 topology). This supports a cytoplasmic, potentially reticular interpretation, but does not establish how sharply ER structure will resolve in chromogenic sections.
Tissue IHC confidence	The tissue profile is labelled Uncertain because antibody staining and RNA expression show low consistency; the listed HPA antibody, HPA003334, also has IHC validation marked Uncertain (HPA: tissue IHC; HPA: antibody validation). Use the reported pattern as a comparison, not proof of antibody specificity.
Isoforms and epitope information	UniProt lists two MOSPD2 isoforms (UniProt Q8NHP6 isoforms). The supplied evidence does not locate the catalog antibody's epitope or show whether it recognizes both isoforms. Do not attribute a staining difference to isoform selectivity without separate antibody-specific evidence.
Q: What should IF/ICC show?	A: ER localization is the supported main location in HPA ICC-IF images, consistent with UniProt's ER membrane annotation (HPA: subcellular; UniProt Q8NHP6 subcellular location). This cross-check concerns localization; IF/ICC staining conditions belong in its separate guide and do not define an IHC protocol.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Expected cells show little or no chromogenic signal.	The reported positive population may be absent from the section, or the routine IHC workflow may have failed.	Confirm the cell population first; appendix glandular and bone marrow hematopoietic cells are reported Medium (HPA: tissue IHC). Then review retrieval, primary dilution and detection controls without assuming a MOSPD2-specific retrieval requirement.
Nuclei dominate the stain.	The pattern disagrees with MOSPD2's ER membrane location (UniProt Q8NHP6 subcellular location).	Compare the no-primary control and inspect whether colour tracks nuclei rather than cytoplasm. Score only a reproducible cellular pattern consistent with the expected compartment; investigate persistent nuclear staining as possible nonspecific signal.
Adipocytes or cardiomyocytes stain strongly.	These cells are reported Not detected, making cross-reactivity or endogenous chromogenic activity possible explanations (HPA: tissue IHC).	Verify cell identity and examine a no-primary control; check the routine endogenous enzyme block if using enzyme-based detection. Interpret disagreement cautiously because HPA rates the tissue profile Uncertain (HPA: Uncertain).
The entire section has diffuse brown background.	Routine blocking, wash or detection background may obscure cellular staining.	Use a no-primary control to separate detection background from antibody-associated colour, then review blocking, washes and primary dilution. Seek a discrete cytoplasmic pattern before comparing the section with HPA's cytoplasmic tissue profile (HPA: tissue IHC).
Two reported positive tissues stain differently.	HPA's Medium calls describe specific cell populations, and its overall tissue IHC reliability is Uncertain (HPA: tissue IHC).	Compare the same named cell types across appropriately controlled sections and record intensity and the fraction of cells stained. Do not treat a difference between whole tissues as an established MOSPD2 expression change.
Signal appears in a reported undetected population but not in reported positive cells.	A reversed pattern is inconsistent with the supplied cell-level IHC observations, though those observations remain provisional (HPA: tissue IHC; HPA: Uncertain).	Recheck slide labels, tissue morphology and detection controls, then reassess specificity with an independent validation approach if the reversal persists. Avoid calling the unexpected stain a confirmed MOSPD2 pattern solely from chromogenic colour.

Source: UniProt Q8NHP6 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing MOSPD2 staining

IHC analysis of MOSPD2 using anti-MOSPD2 antibody (A16695-1). MOSPD2 was detected in a paraffin-embedded section of human brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-MOSPD2 Antibody (A16695-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

A16695-1 · IHCIHC analysis of MOSPD2 using anti-MOSPD2 antibody (A16695-1). MOSPD2 was detected in a paraffin-embedded section of human brain tissue. Heat

IHC analysis of MOSPD2 using anti-MOSPD2 antibody (A16695-1). MOSPD2 was detected in a paraffin-embedded section of human colorectal adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-MOSPD2 Antibody (A16695-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

A16695-1 · IHCIHC analysis of MOSPD2 using anti-MOSPD2 antibody (A16695-1). MOSPD2 was detected in a paraffin-embedded section of human colorectal adenoca

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000130150-MOSPD2/tissue/cerebral+cortex#imid_1150544

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000130150-MOSPD2/tissue/colon#imid_1150478

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000130150-MOSPD2/tissue/liver#imid_1150471

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000130150-MOSPD2/tissue/kidney#imid_1150472

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003334; green = MOSPD2, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for MOSPD2 IHC & IF

🧪Run bone marrow first: hematopoietic cells should stain (HPA: Medium in bone marrow hematopoietic cells). Use heart muscle cardiomyocytes as the negative tissue (HPA: Not detected in cardiomyocytes); on the bone marrow slide, independently identified B and T cells should lack convincing staining (UniProt Q8NHP6: no significant expression in B or T cells).

Positive control tissue: Appendix (Glandular cells, HPA Medium)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MOSPD2 in A-431, U-251MG, U2OS, with annotated localisation: Endoplasmic reticulum (supported) (HPA subcellular).

Technical controls: Include a no-primary secondary-only control, a concentration-matched rabbit IgG isotype control matched to the primary antibody’s clonality, and MOSPD2-knockout material as a biological negative (selected-SKU caption: rabbit primary antibody). Quench endogenous peroxidase in bone marrow before HRP/DAB detection and assess residual background in the no-primary control (selected-SKU caption: HRP/DAB detection; standard IHC practice).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU tissue-IHC caption does not report a fixative (selected-SKU caption: fixative not stated). Paraffin-section staining used heat-mediated EDTA retrieval at pH 8.0; this supports that retrieval condition but does not establish how strongly MOSPD2 depends on retrieval (selected-SKU caption). No supplied evidence establishes whether frozen sections or IF/ICC are easier than paraffin IHC; IF/ICC can assess the supported ER localization, while endogenous peroxidase in bone marrow warrants attention for chromogenic IHC (HPA subcellular: ER supported; selected-SKU caption: HRP/DAB detection; standard IHC practice).

HPA tissue IHC evidence for MOSPD2

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low 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
Appendix	Glandular cells	Medium	Protein (IHC)	HPA →
Bone marrow	Hematopoietic cells	Medium	Protein (IHC)	HPA →
Bronchus	Respiratory epithelial cells	Medium	Protein (IHC)	HPA →
Caudate	Glial cells	Medium	Protein (IHC)	HPA →
Cerebellum	Cells in granular layer	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →
Esophagus	Squamous epithelial cells	Not detected	Protein (IHC)	HPA →
Heart muscle	Cardiomyocytes	Not detected	Protein (IHC)	HPA →
Kidney	Cells in glomeruli	Not detected	Protein (IHC)	HPA →
Liver	Cholangiocytes	Not detected	Protein (IHC)	HPA →

Section 3

Advanced MOSPD2 IHC Tips

Troubleshoot MOSPD2 staining in paraffin sections by checking retrieval, compartment, cell identity, and controls before interpreting chromogenic signal.

How should I adjust retrieval when MOSPD2 staining is weak in paraffin sections?

Use heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A16695-1). This is the retrieval used for the selected antibody’s human brain tissue image (datasheet A16695-1). If staining is weak, first verify that sections were fully deparaffinised and that the retrieval bath reached its intended temperature, then compare a longer retrieval with the original condition on adjacent sections (standard IHC practice). Keep the antibody at the documented 2 μg/ml during that comparison so retrieval is the main variable (datasheet A16695-1). Judge improvement by cytoplasmic staining with preserved morphology and low background, since the tissue staining evidence has uncertain reliability (HPA: tissue IHC reliability Uncertain).

Can I attribute weak MOSPD2 staining to the tissue fixative?

The selected image documents a paraffin-embedded human brain section but does not state its fixative, so MOSPD2-specific fixation sensitivity is unknown (datasheet A16695-1). Do not infer a preferred fixative from the tissue staining profile or protein topology (HPA: tissue IHC reliability Uncertain; UniProt Q8NHP6 topology). Record the actual fixative and fixation duration for each specimen, then compare matched sections with the same pH 8.0 EDTA retrieval and 2 μg/ml antibody condition (datasheet A16695-1; standard IHC practice). If signal varies with processing history, check morphology and an independent tissue control before calling the difference biological (standard IHC practice).

What staining pattern is plausible for MOSPD2 in tissue sections?

Expect predominantly cytoplasmic staining, potentially with a fine reticular pattern, because MOSPD2 is anchored in the endoplasmic reticulum membrane (HPA: cytoplasmic expression in most tissues; UniProt Q8NHP6 subcellular location). Its 1–496 cytoplasmic region precedes a transmembrane segment at 497–518, so a nuclear-only pattern needs independent confirmation (UniProt Q8NHP6 topology). Contacts with lipid droplets and other organelles can change the distribution within the ER, but routine chromogenic sections cannot establish a specific contact site from morphology alone (UniProt Q8NHP6 function; standard IHC interpretation). Compare the stained compartment across adjacent cells and sections, and treat the HPA tissue pattern as provisional because its staining and RNA data have low consistency (HPA: tissue IHC reliability Uncertain).

Could epitope placement explain discordant MOSPD2 staining?

MOSPD2 has 2 annotated isoforms, but the supplied antibody evidence does not identify its epitope or establish which isoforms it detects (UniProt Q8NHP6 isoforms; datasheet A16695-1). The protein contains a CRAL-TRIO domain at 82–239, an MSP domain at 327–445, and a membrane anchor at 497–518 (UniProt Q8NHP6 domains and topology). Ask for an epitope map before attributing discordant staining to isoform loss, and compare independent antibodies or orthogonal expression evidence when available (standard IHC validation practice). Retrieval may expose a masked epitope, but a stronger DAB signal after retrieval does not identify the recognized sequence or prove isoform specificity (standard IHC interpretation).

How can I assess MOSPD2 by IF alongside a cell identity marker?

For IF on tissue sections, pair MOSPD2 with a CD14 marker when testing a monocyte-rich population, because MOSPD2 is highly expressed in CD14-positive monocytes (UniProt Q8NHP6 tissue specificity). Choose spectrally separated fluorophores, placing the weaker signal in a far-red channel if tissue autofluorescence interferes, and include single-label and unstained controls (standard IF practice). MOSPD2 residues 1–496 face the cytoplasm, so permeabilisation is needed for an antibody whose epitope lies there; the selected antibody’s epitope is unspecified (UniProt Q8NHP6 topology; datasheet A16695-1). Optimise permeabilisation for the verified epitope and fixation method, then assess reticular signal and marker overlap without assuming every CD14-positive cell must stain (HPA: ER location supported; standard IF practice).

What should I check if DAB background obscures MOSPD2 staining?

Start with a no-primary control to distinguish detection-system staining from antibody-associated signal, and inspect tissue edges and damaged areas separately (standard IHC practice). The selected image used 10% goat serum, 2 μg/ml primary antibody overnight at 4°C, and an anti-rabbit secondary for 30 minutes at 37°C (datasheet A16695-1). If background remains high, verify peroxidase blocking and washing, then titrate the primary antibody while keeping retrieval and DAB development consistent (standard chromogenic IHC practice). Treat diffuse staining outside the expected cytoplasmic compartment cautiously, especially because independent tissue staining and RNA expression show low consistency (UniProt Q8NHP6 subcellular location; HPA: tissue IHC reliability Uncertain).

How should I score MOSPD2 staining across heterogeneous sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and region before scoring, then record the percentage of cells at each cytoplasmic intensity from 0–3 and calculate an H-score from 0–300 (standard IHC scoring practice; HPA: cytoplasmic expression in most tissues). For sparse infiltrates, report MOSPD2-positive cells per mm² alongside the fraction of eligible cells that stain, so cellular abundance and expression are distinguishable (standard IHC quantification practice). Normalise counts to viable tissue area and percentages to the relevant cell population, applying the same thresholds and imaging conditions across sections (standard IHC quantification practice). Exclude folds, edges, and necrotic areas before comparison, and report staining as assay-defined because HPA tissue reliability is uncertain (standard IHC practice; HPA: tissue IHC reliability Uncertain).

How can I distinguish credible MOSPD2 signal from artefact?

A credible signal should occur in a plausible cytoplasmic compartment and be reproducible across intact cells, consistent with MOSPD2’s ER membrane location (UniProt Q8NHP6 subcellular location; standard IHC interpretation). Monocyte-rich populations are biologically plausible targets, whereas a nuclear-only pattern or staining confined to section edges warrants further validation (UniProt Q8NHP6 tissue specificity and topology; standard IHC interpretation). Compare viable areas with necrotic or damaged areas, inspect a no-primary control for endogenous peroxidase or detection background, and confirm questionable cells with an independent marker when possible (standard chromogenic IHC practice). Do not treat the selected brain image or a single positive region as proof of tissue specificity: HPA reports low tissue specificity and uncertain tissue IHC reliability (datasheet A16695-1; HPA: RNA specificity Low tissue specificity; HPA: tissue IHC reliability Uncertain).

Boster reagents

Best MOSPD2 / Motile sperm domain-containing protein 2 IHC Antibodies

A16695-1 has real IHC data from paraffin-embedded human brain, colorectal adenocarcinoma, placenta, and testicular germ cell tumor sections (catalog IHC captions). No IF images are supplied (catalog image payload).

Real IHC data

Anti-MOSPD2 Antibody ®

Cat # A16695-1

View product →

A16695-1 is listed for IHC in human, mouse, and rat (catalog applications/reactivity). Its own IHC captions document DAB staining in paraffin-embedded human brain, colorectal adenocarcinoma, placenta, and testicular germ cell tumor sections (catalog IHC captions).

Which to pick: For tissue IHC, choose A16695-1 at a listed starting range of 2–5 μg/ml (datasheet: IHC dilution); its own captions show paraffin sections, but do not report the fixative (catalog IHC captions). For IF/ICC, A16695-1 has no listed validation or IF images, so there is no supported IF/ICC pick in this payload (catalog applications and image payload). For cross-species IHC, A16695-1 lists human, mouse, and rat reactivity, while its IHC images document human tissue only (catalog reactivity; catalog IHC captions).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry Q8NHP6 (MSPD2_HUMAN, Motile sperm domain-containing protein 2).
Human Protein Atlas. MOSPD2 tissue IHC expression (reliability: Uncertain).
Human Protein Atlas. MOSPD2 subcellular location (ICC-IF): Localized to the endoplasmic reticulum..
Human Protein Atlas. MOSPD2 antibody validation summary (1 antibodies).
Epstein-Barr virus-encoded microRNA BART22 serves as novel biomarkers and drives malignant transformation of nasopharyngeal carcinoma. Cell death & disease 2022 — PMC9338958.
PubMed PMID:14702039 — UniProt-cited evidence.
PubMed PMID:15489334 — UniProt-cited evidence.
PubMed PMID:17974005 — UniProt-cited evidence.