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MEP1B / Meprin A subunit beta · IHC design guide
Design Immunohistochemistry for MEP1B
Plan MEP1B IHC in paraffin sections around the enterocyte brush-border pattern seen in gastrointestinal tissue (HPA tissue IHC). Use its membrane topology and potential surface shedding to interpret chromogenic staining (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MEP1B 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 | Enterocyte brush border (HPA tissue IHC) | |
| Staining pattern | Membranous staining of GI enterocyte microvilli (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06392-1) | |
| Positive control | Colon+2 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 sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A06392-1); verify before use. | |
| Caveat | Surface shedding may alter membrane staining (UniProt) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 0 isoforms listed; check extracellular vs cytoplasmic epitope (UniProt) |
Section 1
Recommended MEP1B IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with one published MEP1B uterine-tissue IHC protocol (PMC9990359).
| Sample | Paraffin-embedded mouse intestine tissue; fixative not specified (datasheet A06392-1) |
| Fixation | Image fixative and duration unreported (datasheet A06392-1); 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 A06392-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06392-1) |
| Primary antibody | Rabbit anti-MEP1B, 2-5 μg/ml (datasheet A06392-1) |
| Primary incubation | Overnight at 4 °C (datasheet A06392-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06392-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MEP1B-positive staining in enterocytes - Microvilli of colon (HPA tissue IHC: High). HPA tissue profile: Selective expression in brush border membrane in gastrointestinal tract. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A06392-1); the published protocol does not specify retrieval (PMC9990359).
Section 2
What Is the Expected MEP1B Staining Pattern?
MEP1B is a surface protein with an extracellular domain and can be shed (UniProt Q16820 topology). In paraffin IHC, expect strongest staining along enterocyte microvilli in colon, duodenum and small intestine (HPA tissue IHC: High). The selective brush border pattern has Enhanced tissue reliability, with high consistency between staining and RNA data (HPA tissue IHC).
| A crisp apical rim on intestinal enterocytes, strongest at the microvilli. | This matches the reported High microvillar staining in colon, duodenum and small intestine (HPA tissue IHC). Its surface position is consistent with the extracellular domain and membrane anchor (UniProt Q16820 topology). |
| Strong nuclear staining dominates, while the apical enterocyte rim is absent. | Nuclear localisation is unsupported by the reported membrane and secreted locations (UniProt Q16820; HPA subcellular summary). Treat a dominant nuclear pattern as suspect and check controls before assigning it to MEP1B (general IHC practice). |
| Widespread staining appears in cells outside the reported enterocyte pattern. | The HPA result is selective for gastrointestinal brush border (HPA tissue IHC). Unexpected staining may reflect antibody cross-reactivity or endogenous chromogenic activity; a negative reagent control helps separate these possibilities (general IHC practice). |
| Colour coats the section diffusely and obscures cell borders. | This cannot establish the selective microvillar pattern reported by HPA (HPA tissue IHC). Uneven reagent coverage, incomplete blocking or inadequate washing can contribute to diffuse background in chromogenic IHC (general IHC practice). |
| No apical signal appears in an otherwise intact duodenum or small intestine section. | These are reported High enterocyte microvillar sites (HPA tissue IHC). Review tissue orientation and the staining run before calling the sample negative; loss of the visible apical edge can prevent assessment (general IHC practice). |
💡Expected MEP1B appearanceCall the result positive when High, crisp apical microvillar staining marks intestinal enterocytes (HPA tissue IHC); dominant nuclear or widespread nonselective colour is suspect (UniProt Q16820 topology; general IHC practice).
| Tissue and cell selection | Colon, duodenum and small intestine have High enterocyte microvillar staining (HPA tissue IHC). HPA's Not detected entries describe the listed cell types, not every cell in each tissue (HPA tissue IHC). |
| Membrane topology and shedding | Residues 23–652 are extracellular and residues 653–673 span the membrane; surface homodimers may be shed (UniProt Q16820 topology). Extracellular signal therefore needs anatomical and control context (general IHC practice). |
| Antibody evidence | HPA lists HPA029119 as Enhanced for IHC, and tissue staining has Enhanced reliability (HPA antibodies; HPA tissue IHC). These ratings support the reported pattern but do not identify the cause of every unexpected signal (general IHC practice). |
| Retrieval and epitope | No antibody epitope or MEP1B-specific retrieval response is supplied (UniProt Q16820; HPA tissue IHC). If optimising retrieval, compare conditions against a known positive section and controls (general IHC practice). |
| What should IF/ICC show? | Membrane localisation and possible shedding inform interpretation (UniProt Q16820), but HPA supplies no ICC-IF images or main location assignment for MEP1B (HPA subcellular). An IF/ICC pattern is therefore unconfirmed by this evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected intestinal apical rim is absent. | A staining-run failure or an apical border outside the plane of section may hide the reported High microvillar pattern (HPA tissue IHC; general IHC practice). | Inspect morphology and section orientation; repeat alongside a positive intestinal section and suitable reagent controls (general IHC practice). |
| The entire section develops a uniform brown haze. | Blocking, washing or chromogen development may be contributing background (general IHC practice). | Review blocking, washes and development time; compare a negative reagent control before interpreting faint apical colour (general IHC practice). |
| Strong colour appears in cells that lack a matching apical pattern. | Cross-reactivity or endogenous detection activity is possible (general IHC practice); HPA reports selective gastrointestinal brush border staining (HPA tissue IHC). | Compare cellular morphology, the positive tissue pattern and a negative reagent control; reassess antibody specificity if unexpected staining persists (general IHC practice). |
| Predominantly nuclear staining is scored as positive. | A nuclear pattern conflicts with the membrane and secreted locations in the supplied records (UniProt Q16820; HPA subcellular summary). | Withhold a positive call and inspect controls and the apical enterocyte compartment (general IHC practice; HPA tissue IHC). |
| Extracellular colour appears near an apical surface. | Surface MEP1B can be shed by ADAM10 or ADAM17 (UniProt Q16820), but adjacent colour alone cannot establish its identity (general IHC practice). | Assess whether a crisp enterocyte microvillar rim is also present, then compare controls before assigning extracellular signal to MEP1B (HPA tissue IHC; general IHC practice). |
| A reported HPA negative cell type stains weakly. | Not detected applies to the specified cell type and assay observation; it is not a tissue-wide exclusion (HPA tissue IHC). Weak chromogenic background is also possible (general IHC practice). | Identify the stained cell type, compare a negative reagent control and use the intestinal apical pattern as the positive reference (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing MEP1B staining
A06392-1 · IHCIHC analysis of MEP1B using anti-MEP1B antibody (A06392-1). MEP1B was detected in a paraffin-embedded section of mouse intestine tissue. Hea
A06392-1 · IHCIHC analysis of MEP1B using anti-MEP1B antibody (A06392-1). MEP1B was detected in a paraffin-embedded section of rat intestine tissue. Heat
A06392-1 · IFIF analysis of MEP1B using anti-MEP1B antibody (A06392-1). MEP1B was detected in paraffin-embedded section of mouse colon tissue. Heat media
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000141434-MEP1B/tissue/cerebral+cortex#imid_17037271
HPA · Small intestineHuman Protein Atlas IHC, Small intestine. Source: https://www.proteinatlas.org/ENSG00000141434-MEP1B/tissue/small+intestine#imid_17037156
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000141434-MEP1B/tissue/colon#imid_17037189
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000141434-MEP1B/tissue/liver#imid_17037182
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MEP1B IHC & IF
🧪Run duodenum first: enterocyte microvilli should stain strongly (HPA: High in duodenum enterocyte microvilli). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the duodenum slide, non-microvillar areas should show only background staining relative to the brush border (HPA: High in enterocyte microvilli).
Positive control tissue: Colon (Enterocytes - Microvilli, 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 MEP1B; derive a cell-line control from the positive tissue's cell type (Enterocytes - Microvilli) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls (selected-SKU caption: rabbit primary antibody), plus MEP1B knockout tissue if available or a validated immunizing-peptide competition control (standard IHC specificity controls). Quench endogenous peroxidase and check for residual DAB signal in duodenum (selected-SKU caption: HRP/DAB detection; standard chromogenic IHC practice).
⚠️Feasibility: The selected-SKU paraffin-section example used heat retrieval in EDTA at pH 8.0 and 2 μg/ml primary antibody overnight at 4°C (selected-SKU caption); it does not establish that retrieval is required. Its fixative is unreported, and no target-specific fixation window or fixation effect is reported in the supplied evidence (selected-SKU caption: fixative not stated). The supplied evidence does not establish that frozen sections or IF are easier; assess luminal debris and brush-border background against the controls (HPA: High in enterocyte microvilli; standard IHC practice).
HPA tissue IHC evidence for MEP1B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex.). 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 |
|---|---|---|---|---|
| Colon | Enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Duodenum | Enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Small intestine | Enterocytes - Microvilli | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MEP1B IHC Tips
Troubleshoot chromogenic MEP1B IHC in paraffin sections by checking retrieval, epithelial brush border staining, assay background, and compartment specific scoring.
Which retrieval conditions should I start with for MEP1B in paraffin sections?
Use heat mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A06392-1). The selected mouse intestine image used this retrieval before 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A06392-1). If staining is weak, compare matched serial sections while adjusting heating and cooling duration, and include a similarly processed small intestine control (standard IHC practice; HPA: high in small intestine enterocyte microvilli). Judge recovery by crisp apical staining and preserved epithelial morphology rather than diffuse luminal color (HPA: selective brush border expression; standard IHC practice).
How should I troubleshoot a fixation related loss of MEP1B staining?
The selected paraffin section caption does not state its fixative, so target specific fixation sensitivity is unknown (datasheet A06392-1). Record the fixative, fixation duration, and processing history for each specimen, then compare matched sections processed under documented conditions (standard IHC practice). Keep EDTA retrieval at pH 8.0 and the primary concentration at 2 μg/ml while investigating fixation, so changes in staining have fewer competing explanations (datasheet A06392-1; standard IHC practice). Assess epithelial preservation alongside signal, using apical enterocyte staining in intestine as the expected pattern (HPA: high in small intestine enterocyte microvilli; standard IHC practice).
Where should convincing MEP1B staining appear in an intestinal section?
Look first for an apical brush border pattern in intestinal enterocytes, especially their microvilli (HPA: selective gastrointestinal brush border expression; HPA: high in small intestine enterocyte microvilli). MEP1B has an extracellular region at residues 23–652 and a membrane spanning segment at 653–673, consistent with surface staining (UniProt Q16820 topology). Surface shedding by ADAM10 and ADAM17 can also place protein outside the membrane, so extracellular color needs anatomical and control based review (UniProt Q16820 subcellular location; standard IHC practice). Compare intact apical borders with adjacent luminal material and avoid calling diffuse deposits equivalent to enterocyte membrane expression (HPA: brush border profile; standard IHC practice).
Could epitope placement explain staining differences between MEP1B antibodies?
Check the catalog antibody’s stated immunogen or mapped epitope before comparing staining patterns; the supplied image caption does not identify its epitope (datasheet A06392-1; standard IHC practice). The supplied UniProt record lists 0 isoforms, a signal peptide at 1–22, and a propeptide at 23–61 (UniProt Q16820 processing and isoforms). Its extracellular region contains multiple annotated glycosylation sites, so epitope accessibility may vary with processing and retrieval (UniProt Q16820 topology and glycosylation; standard IHC practice). Compare antibodies on matched paraffin sections with the same retrieval and assess agreement at the apical enterocyte border before attributing disagreement to a protein variant (HPA: brush border profile; standard IHC practice).
How can I investigate MEP1B localisation with multiplex IF after chromogenic IHC?
Treat multiplex IF as a separate assay requiring its own validation, since the selected antibody evidence describes chromogenic staining of a paraffin mouse intestine section (datasheet A06392-1; standard IF practice). Pair MEP1B with a validated enterocyte marker and assess whether signal follows the apical microvilli rather than simply overlapping intestinal tissue (HPA: high in enterocyte microvilli; standard IF practice). Inspect unstained tissue for autofluorescence, then choose spectrally separated fluorophores and acquisition settings that distinguish that signal from the marker channels (standard IF practice). Set permeabilisation according to the antibody’s mapped epitope: the extracellular region is 23–652, whereas the cytoplasmic tail is 674–701 (UniProt Q16820 topology; standard IF practice).
What should I check when DAB appears across the whole section?
Start with a no primary control and check whether color persists after the secondary antibody and DAB steps (standard IHC practice). Block endogenous peroxidase for an HRP and DAB workflow, and evaluate whether the block removes tissue associated color without weakening the positive control (standard IHC practice). The selected image used 10% goat serum blocking, 2 μg/ml rabbit primary overnight at 4°C, and peroxidase conjugated secondary for 30 minutes at 37°C (datasheet A06392-1). If background remains, review washes, antibody concentration, and DAB development while preserving a crisp enterocyte brush border signal (standard IHC practice; HPA: brush border profile).
How should I score MEP1B staining without counting luminal deposits? ⚠ ANSWER MARKED FOR VERIFICATION
Define enterocyte apical membrane as the scoring compartment before reviewing slides, because gastrointestinal brush border staining is the reported tissue pattern (HPA: selective brush border expression; standard IHC practice). Report the percentage of enterocytes with apical staining and, if intensity is reproducible, an H-score from 0–300 using intensity grades 0–3 (standard IHC practice). Normalise counts to evaluable enterocytes or intact epithelial length, and report any area based density per mm² only for a predefined epithelial region (standard IHC practice). Exclude detached luminal material, necrotic areas, and section edges using the same documented criteria for every specimen (standard IHC practice).
How do I distinguish true MEP1B signal from staining artefact?
Favor reproducible apical staining in intact enterocytes of intestine, where microvillar signal is reported as high (HPA: high in small intestine enterocyte microvilli; standard IHC practice). Compare this pattern with a no primary control and with a tissue region expected to show little signal, such as adipocytes in adipose tissue (standard IHC practice; HPA: adipocytes not detected). Treat predominantly nuclear or diffuse stromal color as discordant with the annotated membrane and secreted localisation until independently verified (UniProt Q16820 subcellular location; standard IHC practice). Check section edges, necrotic debris, and persistent DAB color after primary omission for processing artefact or endogenous peroxidase activity (standard IHC practice).
Boster reagents
Best MEP1B / Meprin A subunit beta IHC Antibodies
A06392-1 has IHC images from paraffin-embedded mouse and rat intestine and IF images from paraffin-embedded mouse and rat colon; Human reactivity is listed (catalog IHC/IF image captions; catalog reactivity).
Real IHC data
Anti-MEP1B Antibody ®
Cat # A06392-1
A06392-1 has IHC data from paraffin-embedded mouse and rat intestine (IHC image captions). The same SKU has IF data from paraffin-embedded mouse and rat colon and lists Human, Mouse, and Rat reactivity (IF image captions; catalog reactivity).
Which to pick: For tissue IHC, choose A06392-1: its rabbit primary was shown on paraffin-embedded mouse and rat intestine at 2 μg/ml after EDTA pH 8 retrieval; the fixative is unreported (IHC image captions). For IF, A06392-1 has mouse and rat colon images at 5 μg/ml; ICC validation and clonality are unreported (IF image captions; catalog applications and clone field). For cross-species studies, A06392-1 lists Human, Mouse, and Rat reactivity, while the supplied IHC/IF images show Mouse and Rat tissue (catalog reactivity; IHC/IF image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q16820 (MEP1B_HUMAN, Meprin A subunit beta).
- Human Protein Atlas. MEP1B tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. MEP1B subcellular location (ICC-IF): Membrane, Secreted.
- Human Protein Atlas. MEP1B antibody validation summary (1 antibodies).
- Uterine luminal-derived extracellular vesicles: potential nanomaterials to improve embryo implantation. Journal of nanobiotechnology 2023 — PMC9990359.
- Integrating LCM-Based Spatio-Temporal Transcriptomics Uncovers Conceptus and Endometrial Luminal Epithelium Communication that Coordinates the Conceptus Attachment in Pigs. International journal of molecular sciences 2021 — PMC7866100.
- Targeted fibrillar nanocarbon RNAi treatment of acute kidney injury. Science translational medicine 2016 — PMC5004247.
- PubMed PMID:9288916 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:8262185 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MEP1B (Meprin A subunit beta). Expected localisation Enterocyte brush border (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06392-1). Validated IHC antibody: A06392-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Colon (Enterocytes - Microvilli); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse intestine tissue; fixative not specified (datasheet A06392-1); 2. Fixation: Image fixative and duration unreported (datasheet A06392-1); 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 A06392-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06392-1); 8. Primary antibody: Rabbit anti-MEP1B, 2-5 μg/ml (datasheet A06392-1); 9. Primary incubation: Overnight at 4 °C (datasheet A06392-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A06392-1); verify before use.; Caveat: Surface shedding may alter membrane staining (UniProt); Isoform / epitope: 0 isoforms listed; check extracellular vs cytoplasmic epitope (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q16820, HPA tissue IHC · Enhanced, HPA ICC-IF · Membrane, Secreted, Datasheet A06392-1, PMC9990359. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MEP1B/immunohistochemistry-ihc-guide.