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MTNR1B / Melatonin receptor type 1B · IHC design guide
Design Immunohistochemistry for MTNR1B
Plan MTNR1B staining in paraffin sections using cerebral cortex neurons, retinal nerve fibers and hippocampal glia as high-staining reference sites (HPA tissue IHC). Interpret the observed cytoplasmic pattern (HPA tissue IHC) alongside its cell-membrane topology (UniProt) and the reported off-target staining concern (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MTNR1B 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 tissue signal (HPA tissue IHC); cell membrane (UniProt) | |
| Staining pattern | Cytoplasmic neuronal processes and placental decidual cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A30844) | |
| Positive control | Caudate+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A30844) | |
| Caveat | Presumed off-target staining warrants careful interpretation (HPA tissue IHC) | |
| Regulation | No expression regulator specified (UniProt) | |
| Isoform / epitope | No isoforms annotated; epitope side matters across 7 transmembrane segments (UniProt) |
Section 1
Recommended MTNR1B IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses heat-mediated Tris-EDTA retrieval at pH 9.0 (datasheet A30844). The published options below cover human glioma and sheep CNS tissue (PMC13263238; PMC3608070).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A30844) |
| Fixation | Image fixative and duration unreported (datasheet A30844); 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 A30844); 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-MTNR1B, 1:50 recommended; image 1:200 (datasheet A30844) |
| 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 | MTNR1B-positive staining in glial cells of caudate (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in neuronal processes in CNS and retina, and decidual cells in placenta. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA retrieval at pH 9.0 for the catalog antibody (datasheet A30844); neither included article specifies an MTNR1B retrieval method (PMC13263238; PMC3608070).
Section 2
What Is the Expected MTNR1B Staining Pattern?
MTNR1B is a 7-pass cell-membrane receptor (UniProt P49286 topology). In paraffin-section IHC, expect staining in neuronal processes in CNS and retina, and decidual cells in placenta (HPA: tissue IHC, Supported). HPA also reports high staining in specified neuronal and glial cells; its Supported assessment notes presumed off-target staining was disregarded and that RNA data do not corroborate the antibody pattern (HPA: tissue IHC).
| Staining follows neuronal processes or the retinal nerve fiber layer, with little nuclear signal (HPA: tissue IHC; UniProt P49286 topology). | This fits the reported cytoplasmic-appearing process pattern for a membrane receptor (HPA: tissue IHC; UniProt P49286 subcellular location). Compare the named cell compartment and tissue; a continuous cell outline is not required to recognize process staining in a section (general IHC practice). |
| Signal is mainly nuclear in cells expected to stain (HPA: tissue IHC). | Nuclear dominance conflicts with the cell-membrane assignment and reported process or decidual-cell staining (UniProt P49286 subcellular location; HPA: tissue IHC). Treat it as suspect until controls exclude nonspecific staining or a detection artefact (general IHC practice). |
| Strong staining appears in adipocytes or adrenal glandular cells (HPA: Not detected in those cells). | These are poor matches to the HPA tissue pattern, so review cell identification and background before calling them MTNR1B-positive (HPA: tissue IHC). Cross-reactivity or endogenous detection activity can mimic a positive chromogenic signal (general IHC practice). |
| Color is diffuse across cells and surrounding tissue, including areas without a recognizable cell pattern (general IHC practice). | Diffuse staining cannot establish the reported neuronal-process, retinal nerve-fiber, or decidual-cell pattern (HPA: tissue IHC). Compare a no-primary control and inspect blocking, washes, and detection chemistry for background (general IHC practice). |
| No signal appears in a section containing retinal nerve fiber layer or cortical neuronal cells (HPA: High in those locations). | First check that the expected cells are present and the detection run worked (general IHC practice). An absent signal conflicts with the HPA IHC observations, but it cannot by itself prove absent MTNR1B: antibody and section conditions still need review (HPA: tissue IHC; general IHC practice). |
💡Expected MTNR1B appearanceCall positive when chromogenic staining follows neuronal processes or retinal nerve fibers, or appears in decidual cells, at the reported cell-specific intensity; dominant nuclear staining or broad staining of HPA-negative cells is suspect (HPA: tissue IHC; UniProt P49286 subcellular location).
| Compartment and topology (UniProt P49286 topology; HPA: tissue IHC) | Seven membrane-spanning segments support a membrane-associated interpretation; the HPA section profile describes cytoplasmic-appearing neuronal processes (UniProt P49286 topology; HPA: tissue IHC). Interpret section morphology in that context rather than requiring a sharp membrane rim (general IHC practice). |
| Tissue and cell context (HPA: tissue IHC) | High staining is reported in cortical neuronal cells, caudate and hippocampal glial cells, and retinal nerve fiber layer; placenta decidual cells show medium staining (HPA: tissue IHC). Score the named cells, since a whole-section average can obscure their pattern (general IHC practice). |
| Evidence strength (HPA: IHC Supported; HPA: tissue IHC) | The listed antibody has Supported IHC status, with external characterization backing the observed pattern; HPA also says RNA is not detected and presumed off-target staining was disregarded (HPA: tissue IHC; HPA074024 IHC Supported). Use morphology and controls when judging an unexpected positive (general IHC practice). |
| Extracellular terminus and epitope uncertainty (UniProt P49286 topology and glycosylation) | Residues 1–42 are extracellular and residue 4 is annotated as glycosylated (UniProt P49286 topology and glycosylation). The supplied sources do not map this antibody's epitope or report MTNR1B-specific retrieval or fixation sensitivity; do not infer a retrieval effect from topology alone. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive tissue is blank (HPA: High in retinal nerve fiber layer and cortical neuronal cells). | The section may lack the relevant cells, or the IHC run may have failed (HPA: tissue IHC; general IHC practice). | Confirm the cell compartment on the counterstain and check a working positive control, primary-antibody step, retrieval record, and chromogenic detection (general IHC practice). No MTNR1B-specific retrieval setting is established by these sources. |
| Signal is predominantly nuclear (UniProt P49286: cell membrane). | A dominant nuclear pattern is inconsistent with the reported localization; nonspecific staining or detection background is possible (UniProt P49286 subcellular location; HPA: tissue IHC; general IHC practice). | Compare a no-primary control, inspect background distribution, and reassess the cellular compartment before scoring positive (general IHC practice). |
| Adipocytes or adrenal glandular cells stain strongly (HPA: Not detected in those cells). | Cell misidentification, cross-reactivity, or endogenous detection activity may explain staining outside the reported pattern (HPA: tissue IHC; general IHC practice). | Verify morphology and compare a no-primary control; apply suitable endogenous-activity blocking for the detection system if control staining warrants it (general IHC practice). |
| The whole section has diffuse chromogen (general IHC practice). | Background from primary or detection reagents, inadequate blocking, or insufficient washing can obscure a cell-specific pattern (general IHC practice). | Review no-primary control, blocking and wash steps, then reassess whether staining resolves into HPA-reported cells and processes (general IHC practice; HPA: tissue IHC). |
| Placental decidual staining looks weaker than CNS staining (HPA: Medium in decidual cells; High in listed CNS cells). | Different reported HPA levels can produce this comparison without indicating assay failure (HPA: tissue IHC). | Judge decidual cells against their own expected medium pattern and local background; avoid applying a CNS intensity threshold to placenta (HPA: tissue IHC; general IHC practice). |
| Can ICC-IF confirm the IHC pattern? (HPA: subcellular summary Membrane). | HPA lists membrane as its ICC-IF summary but provides no ICC-IF images or ICC validation status for the listed antibody (HPA: subcellular; HPA074024 antibody record). | Treat the ICC-IF summary as limited context. Base this section's positive call on the Supported tissue-IHC pattern and appropriate IHC controls (HPA: tissue IHC; HPA074024 IHC Supported; general IHC practice). |
Experiment evidence
IHC & IF images showing MTNR1B staining
A30844 · IHCImmunohistochemical analysis of paraffin-embedded human liver cancer. 1, Antibody was diluted at 1:200(4° overnight). 2, Tris-EDTA,pH9.0 was
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000134640-MTNR1B/tissue/cerebral+cortex#imid_20639027
HPA · RetinaHuman Protein Atlas IHC, Retina. Source: https://www.proteinatlas.org/ENSG00000134640-MTNR1B/tissue/retina#imid_1233
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000134640-MTNR1B/tissue/colon#imid_20638937
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000134640-MTNR1B/tissue/liver#imid_20638946
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MTNR1B IHC & IF
🧪Run cerebral cortex first and expect staining in neuronal cells (High; HPA: cerebral cortex neuronal cells). Use adipose tissue as the negative tissue, expecting adipocytes to remain at background (HPA: adipose adipocytes not detected); on the cortex slide, use cells without detectable staining as an internal background reference without assigning them a negative cell identity.
Positive control tissue: Caudate (Glial 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 MTNR1B; derive a cell-line control from the positive tissue's cell type (Glial cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary (secondary-only) control, a host-species- and clonality-matched isotype control, and a target-specific biological negative such as knockout tissue or peptide-blocked antibody where available (standard IHC practice). Block endogenous peroxidase for chromogenic detection in cortex, and assess tissue autofluorescence if evaluating IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected A30844 paraffin-section caption does not report a fixative (selected-SKU tissue-IHC caption). That caption uses Tris–Tris-EDTA pH 9.0 retrieval and a 1:200 primary antibody dilution overnight at 4 °C, so retrieval should be checked when reproducing its chromogenic IHC conditions (selected-SKU tissue-IHC caption). The supplied evidence does not establish whether frozen sections or IF are easier; evaluate membrane-associated signal against background and tissue autofluorescence (UniProt P49286: cell membrane; HPA: membrane summary; standard IHC/IF practice).
HPA tissue IHC evidence for MTNR1B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Presumed off target binding observed and disregarded. External characterization data supports antibody staining pattern while RNA data does not.). 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 |
|---|---|---|---|---|
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | High | Protein (IHC) | HPA → |
| Retina | Nerve fiber layer | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | Medium | 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 | Glandular 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 MTNR1B IHC Tips
Troubleshoot MTNR1B staining in paraffin sections by checking retrieval, compartment, cell type and controls before assigning biological meaning.
Which retrieval condition should I try first for MTNR1B in paraffin sections?
Start with heat-mediated Tris-EDTA at pH 9.0 for MTNR1B paraffin-section IHC (datasheet A30844). The selected image used a 1:200 primary antibody dilution at 4°C overnight, so keep that condition consistent while assessing retrieval (caption A30844). Compare retrieved sections with an otherwise matched section processed without retrieval to judge whether heating improves specific staining (general IHC practice). If staining remains weak, adjust heating duration cautiously while monitoring tissue integrity and background (general IHC practice). Document each change, because a stronger chromogenic signal alone does not establish MTNR1B specificity (general IHC practice).
How should I troubleshoot weak staining when fixation history is uncertain?
MTNR1B-specific sensitivity to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (caption A30844). Record the specimen’s fixative and fixation duration when available, and compare sections with similar processing histories before changing antibody conditions (general IHC practice). Inspect morphology and staining across the section, since uneven preservation can complicate chromogenic IHC interpretation (general IHC practice). Use the documented Tris-EDTA pH 9.0 retrieval and 1:200 primary dilution as starting conditions (datasheet A30844; caption A30844). Do not attribute a staining difference to MTNR1B-specific fixation sensitivity without a controlled comparison (general IHC practice).
Should MTNR1B staining appear at the membrane or in the cytoplasm?
MTNR1B is annotated as a cell-membrane receptor with 7 transmembrane segments (UniProt P49286 subcellular location and topology). Assess crisp membrane-associated staining, while allowing for staining in fine cellular processes when anatomy supports that interpretation (UniProt P49286 subcellular location; HPA tissue IHC profile). HPA describes cytoplasmic expression in neuronal processes in CNS and retina, plus decidual cells in placenta (HPA tissue IHC profile). Resolve apparently diffuse cytoplasmic signal against cellular morphology and matched controls before calling it positive (general IHC practice). Strong nuclear-only or structureless staining conflicts with the annotated location and warrants a specificity check (UniProt P49286 subcellular location; general IHC practice).
How does epitope position affect interpretation of MTNR1B staining?
First obtain the catalog antibody’s immunogen or epitope information; the supplied caption does not identify that region (caption A30844). MTNR1B has 7 membrane-spanning segments, an extracellular N terminus at residues 1–42, and a cytoplasmic C terminus at 309–362 (UniProt P49286 topology). Its annotated glycosylation site is residue 4, so epitope accessibility may depend on which region the antibody recognizes; that effect has not been demonstrated here (UniProt P49286 glycosylation and topology; general IHC practice). No isoforms are annotated in the supplied record (UniProt P49286 isoforms). Compare staining with a second, independently mapped epitope when available (general IHC practice).
How can I adapt the IHC findings to multiplex MTNR1B immunofluorescence?
Treat IF/ICC as a separate assay: the selected 1:200 condition and Tris-EDTA pH 9.0 retrieval come from paraffin-section chromogenic IHC (caption A30844; datasheet A30844). Multiplex MTNR1B with a validated marker for the expected neuronal cell type or process, and assess overlap within the same anatomical structures (HPA tissue IHC profile; general IF practice). Choose fluorophores after checking tissue autofluorescence in an unstained section, particularly in the tissue being examined (general IF practice). Once the antibody epitope is mapped, test permeabilisation for a cytoplasmic epitope and consider omitting it for an accessible extracellular epitope (UniProt P49286 topology; general IF practice). Include single-stain and secondary-only controls (general IF practice).
What should I check when MTNR1B chromogenic staining looks widespread?
Inspect a no-primary control and assess endogenous peroxidase activity before attributing widespread DAB signal to MTNR1B (general IHC practice). Include a peroxidase-blocking step and verify that the chromogen reaction is stopped consistently across sections (general IHC practice). The selected image used primary antibody at 1:200 overnight at 4°C and secondary antibody at 1:200 for 45 minutes at room temperature (caption A30844). If background persists, titrate antibody exposure and review blocking and washing while preserving a matched control section (general IHC practice). Treat unexpected staining cautiously because HPA reports presumed off-target binding that it disregarded (HPA tissue IHC reliability description).
How should I score MTNR1B staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical compartment and cell population before scoring, then apply the same criteria to every section (general IHC practice). For cellular staining, record percent-positive cells and intensity, or calculate an H-score on a 0–300 scale (general IHC practice). For process-rich regions, measure positive area or signal density per mm² within a defined region of interest (general IHC practice). Normalise to the number of eligible cells or analyzed tissue area, and exclude folds, edges and necrotic regions consistently (general IHC practice). Report membrane-associated and process-associated staining separately when morphology permits (UniProt P49286 subcellular location; HPA tissue IHC profile; general IHC practice).
What evidence distinguishes true MTNR1B signal from staining artefact?
Prioritise reproducible staining in plausible cells and structures, including neuronal processes in CNS or retina and decidual cells in placenta (HPA tissue IHC profile). Check whether the signal fits a membrane receptor rather than appearing exclusively nuclear or as diffuse, structureless precipitate (UniProt P49286 subcellular location; general IHC practice). Reject edge accentuation, necrotic-area staining and signal retained in no-primary or peroxidase controls as evidence of specific binding (general IHC practice). HPA rates its tissue pattern Supported but reports presumed off-target binding and RNA that was not detected, so its staining alone cannot settle specificity (HPA tissue IHC reliability description; HPA RNA specificity). Seek concordance with an independent antibody or orthogonal assay (general IHC practice).
Boster reagents
Best MTNR1B / Melatonin receptor type 1B IHC Antibodies
A30844 is a human-reactive anti-MTNR1B antibody with IHC data from paraffin-embedded human liver cancer (catalog: A30844; image caption: A30844).
Real IHC data
Anti-MTR1B MTNR1B Antibody
Cat # A30844
A30844 lists IHC among its applications and human reactivity (catalog: A30844). Its IHC image shows paraffin-embedded human liver cancer (image caption: A30844).
Which to pick: For tissue IHC, choose A30844: it is rabbit polyclonal, lists human reactivity, and has an IHC image from paraffin-embedded human liver cancer (catalog: A30844; image caption: A30844). The caption reports Tris-EDTA pH 9.0 retrieval and a 1:200 primary dilution overnight at 4°C; the fixative is unreported (image caption: A30844). No SKU in this payload lists IF/ICC or nonhuman reactivity, so there is no supported IF/ICC or cross-species pick (catalog: A30844 applications/reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P49286 (MTR1B_HUMAN, Melatonin receptor type 1B).
- Human Protein Atlas. MTNR1B tissue IHC expression (reliability: Supported).
- Human Protein Atlas. MTNR1B subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. MTNR1B antibody validation summary (1 antibodies).
- Melatonin Receptor Expression in Primary Uveal Melanoma. International journal of molecular sciences 2024 — PMC11354273.
- Hierarchical Disruption of the Tryptophan-Melatonin Axis Contributes to Glioma Progression Through AKT/ERK/STAT3 Signalling. Journal of cellular and molecular medicine 2026 — PMC13263238.
- Gene and protein patterns of potential prion-related markers in the central nervous system of clinical and preclinical infected sheep. Veterinary research 2013 — PMC3608070.
- Expression and role of melatonin membrane receptors in the hypothalamic-pituitary-testicular axis of Tibetan sheep in a plateau pastoral area. PloS one 2023 — PMC10599587.
- PubMed PMID:7568007 — UniProt-cited evidence.
- PubMed PMID:10696804 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MTNR1B (Melatonin receptor type 1B). Expected localisation Cytoplasmic tissue signal (HPA tissue IHC); cell membrane (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A30844). Validated IHC antibody: A30844 at 1:50 recommended; image 1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Glial cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A30844); 2. Fixation: Image fixative and duration unreported (datasheet A30844); 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 A30844); 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-MTNR1B, 1:50 recommended; image 1:200 (datasheet A30844); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A30844); Caveat: Presumed off-target staining warrants careful interpretation (HPA tissue IHC); Isoform / epitope: No isoforms annotated; epitope side matters across 7 transmembrane segments (UniProt); Regulation: No expression regulator specified (UniProt). Evidence: UniProt P49286, HPA tissue IHC · Supported, HPA ICC-IF · Membrane, Datasheet A30844, PMC13263238, PMC3608070. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MTNR1B/immunohistochemistry-ihc-guide.