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MTRR / Methionine synthase reductase · IHC design guide
Design Immunohistochemistry for MTRR
Plan MTRR IHC in paraffin sections around its cytoplasmic tissue pattern (HPA tissue IHC). Use the catalog antibody’s 2–5 μg/ml IHC range (datasheet A01401-1), with goblet cells and round or early spermatids as high-staining reference populations (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MTRR 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 in tissue sections (HPA tissue IHC) | |
| Staining pattern | Widespread cytoplasmic staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01401-1) | |
| Positive control | Bronchus+4 more · see all | |
| Negative control | Adipose tissue+2 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) | |
| Caveat | Adipocytes may lack detectable staining (HPA tissue IHC) | |
| Regulation | Stimulus-linked regulation is not established (UniProt) | |
| Isoform / epitope | 2 isoforms (A/B); epitope coverage is unknown (UniProt) |
Section 1
Recommended MTRR IHC & IF Protocols
The catalog antibody has a datasheet IHC-P protocol (datasheet A01401-1). Three published MTRR IHC protocols provide tissue-specific examples (PMC6462347; PMC8703036; PMC7116358).
| Sample | Paraffin-embedded human stomach cancer tissue; fixative not specified (datasheet A01401-1) |
| Fixation | Image fixative and duration unreported (datasheet A01401-1); 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 A01401-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01401-1) |
| Primary antibody | Rabbit anti-MTRR, 2-5 μg/ml (datasheet A01401-1) |
| Primary incubation | Overnight at 4 °C (datasheet A01401-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01401-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MTRR-positive staining in goblet cells of bronchus (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A01401-1). The published protocols use citrate or trypsin with their respective antibodies (PMC6462347; PMC8703036; PMC7116358).
Section 2
What Is the Expected MTRR Staining Pattern?
MTRR should appear predominantly in the cytoplasm of many cell types (UniProt Q9UBK8: cytoplasm; HPA tissue IHC: ubiquitous cytoplasmic expression). It has no transmembrane segment (UniProt Q9UBK8 topology). Strong reference patterns include bronchial goblet cells, nasopharyngeal ciliated cell bodies and round or early spermatids (HPA tissue IHC: High). HPA rates the tissue IHC pattern Supported, with medium consistency between staining and RNA expression (HPA tissue IHC: reliability).
| Cytoplasmic staining in bronchial goblet cells or round or early spermatids, with recognizable tissue structure (HPA tissue IHC: High in these cells). | This matches the reported compartment and cell types (UniProt Q9UBK8: cytoplasm; HPA tissue IHC: High). Judge intensity against a positive control processed in the same run (standard IHC practice); a dark deposit alone does not establish specificity. |
| Predominantly nuclear or sharply surface-restricted chromogen in an IHC section, with little cytoplasmic staining. | That is discordant with the tissue IHC profile (HPA tissue IHC: ubiquitous cytoplasmic expression). UniProt reports no transmembrane segment (UniProt Q9UBK8 topology). Review morphology and controls before interpreting it as MTRR; HPA's uncertain nucleoplasmic ICC-IF signal does not validate a nuclear-only IHC pattern (HPA subcellular). |
| Strong staining in adipocytes, oral squamous epithelial cells or ovarian stromal cells, especially when expected positive cells are faint. | HPA reports these specific cell populations as Not detected (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity, then compare with an appropriate no-primary control (standard IHC practice). These observations do not make every cell in those organs a negative control. |
| Diffuse brown haze across cells, stroma and empty areas, without a readable cytoplasmic pattern. | This does not reproduce the reported cell-associated cytoplasmic profile (HPA tissue IHC). Check background in a no-primary control, blocking, washes and chromogen development (standard IHC practice). Haze by itself cannot identify which step caused the problem. |
| No visible staining in bronchial goblet cells or nasopharyngeal ciliated cell bodies (HPA tissue IHC: High in these cells). | Treat the run as potentially unsuccessful before calling the sample MTRR-negative. Verify that the expected cells are present and assess a concurrently processed positive control and detection controls (standard IHC practice). HPA's Supported rating is not a guarantee for every specimen (HPA tissue IHC: reliability). |
💡Expected MTRR appearanceCall an IHC result concordant when clear cytoplasmic chromogen marks expected cells, potentially strongly in bronchial goblet cells or round or early spermatids (UniProt Q9UBK8: cytoplasm; HPA tissue IHC: High); isolated nuclear-only staining or diffuse haze warrants control review (HPA tissue IHC: cytoplasmic profile; standard IHC practice).
| Cell type and comparison tissue | Use cell-level references: HPA reports High staining in bronchial goblet cells and Medium staining in stomach glandular cells, while adipocytes are Not detected (HPA tissue IHC). UniProt reports MTRR in all tissues tested, particularly abundant in skeletal muscle (UniProt Q9UBK8: tissue specificity). Tissue-wide expression does not imply equal staining in every cell. |
| Antibody evidence | The listed rabbit polyclonal HPA038113 has IHC Supported status (HPA antibodies: HPA038113). HPA describes medium consistency between tissue staining and RNA expression (HPA tissue IHC: reliability). Use that evidence to choose controls; it does not independently prove that an unexpected deposit is specific. |
| Protein topology and processing | MTRR has no transmembrane segment, signal peptide or propeptide, and its listed chain spans residues 1–698 (UniProt Q9UBK8 topology and processing). These annotations fit a cytoplasmic interpretation (UniProt Q9UBK8: cytoplasm); they do not establish an antibody epitope or a target-specific antigen retrieval condition. |
| Isoforms and epitope coverage | UniProt lists isoforms A and B (UniProt Q9UBK8: isoforms). The supplied evidence does not locate the antibody epitope or show which isoforms it detects. Do not infer that a regional staining difference reflects one isoform without separate validation. |
| IF/ICC Q: Is nuclear fluorescence an expected MTRR pattern? | A: Cytosol is supported, while nucleoplasm and intermediate filaments are uncertain in HPA ICC-IF (HPA subcellular). Interpret fluorescence with that qualification; this IHC-P section does not establish a nuclear IHC target pattern or an IF/ICC protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells show no chromogen. | The section may lack the relevant cells, or the IHC detection run may have failed (standard IHC practice). | Confirm cell identity on the counterstain, then check a same-run positive tissue and detection controls (standard IHC practice). Bronchial goblet cells are a reported High reference (HPA tissue IHC). |
| A positive control is weak while background stays low. | Working antibody concentration or retrieval conditions may be unsuitable for this run (standard IHC practice); no MTRR-specific fixation sensitivity is supplied. | Check the catalog antibody's IHC-P instructions and titrate against the positive control (standard IHC practice). Do not assign a target-specific fixation or retrieval effect from HPA staining levels. |
| Brown signal appears in cells HPA lists as Not detected. | Cross-reactivity or endogenous detection activity is possible (standard IHC practice); HPA alone cannot distinguish them. | Compare with a no-primary control and review detection blocking (standard IHC practice). Evaluate the named cells, such as adipocytes, rather than labeling the entire organ negative (HPA tissue IHC). |
| Nuclei dominate and cytoplasm is faint. | The pattern differs from reported tissue IHC localization (HPA tissue IHC: ubiquitous cytoplasmic expression); the source of the nuclear deposit is unresolved. | Inspect the no-primary control, counterstain and tissue morphology, then repeat alongside a cytoplasmic positive control if needed (standard IHC practice). Do not transfer uncertain ICC-IF nucleoplasmic localization directly to IHC (HPA subcellular). |
| Uniform haze obscures cell boundaries. | Excess background from blocking, washing or chromogen development is possible (standard IHC practice). | Use the no-primary control to assess background, then adjust the implicated workflow step and reassess cytoplasmic signal in expected cells (standard IHC practice; HPA tissue IHC: cytoplasmic profile). |
| A weak cell population is called negative because a High reference stains strongly. | Expected intensity differs among reported cells: hepatocytes are Low, while bronchial goblet cells are High (HPA tissue IHC). | Score the named cell population and its own control context; record faint cytoplasmic staining separately from absence of staining (standard IHC practice). Avoid applying a High-reference threshold to a Low-reference population (HPA tissue IHC). |
Experiment evidence
IHC & IF images showing MTRR staining
A01401-1 · IHCIHC analysis of MTRR using anti-MTRR antibody (A01401-1). MTRR was detected in a paraffin-embedded section of human stomach cancer tissue. H
A01401-1 · IFIF analysis of MTRR using anti-MTRR antibody (A01401-1). MTRR was detected in a paraffin-embedded section of human stomach cancer tissue. He
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000124275-MTRR/tissue/cerebral+cortex#imid_9941232
HPA · LungHuman Protein Atlas IHC, Lung. Source: https://www.proteinatlas.org/ENSG00000124275-MTRR/tissue/lung#imid_9941119
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000124275-MTRR/tissue/colon#imid_9941166
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000124275-MTRR/tissue/liver#imid_9941159
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA038113; green = MTRR, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA038113; green = MTRR, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MTRR IHC & IF
🧪Run bronchus first and look for staining in goblet cells (HPA: High in bronchus goblet cells); use adipose tissue as the negative comparator, focusing on adipocytes (HPA: Not detected in adipocytes). On the bronchus slide, cells without visible signal should remain at background chromogen levels; HPA does not identify a verified negative bronchial cell type (HPA: bronchus goblet-cell row).
Positive control tissue: Bronchus (Goblet cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MTRR in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (uncertain), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, an isotype-matched rabbit IgG control appropriate to the primary antibody, and an MTRR-knockout biological negative if available (caption: rabbit anti-MTRR antibody; standard IHC controls). Quench endogenous peroxidase for the DAB assay and check goblet-cell mucus for nonspecific staining (caption: peroxidase/DAB detection; HPA: High in bronchus goblet cells).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A01401-1 paraffin-section caption does not state the fixative (selected-SKU caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required for bronchus; frozen sections and IF/ICC are not shown to be easier by the supplied evidence (selected-SKU caption: stomach cancer tissue IHC; HPA: ICC-IF images). Assess mucus-associated background when scoring bronchial goblet cells (HPA: High in bronchus goblet cells; standard IHC practice).
HPA tissue IHC evidence for MTRR
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Bronchus | Goblet cells | High | Protein (IHC) | HPA → |
| Nasopharynx | Ciliated cells (cell body) | High | Protein (IHC) | HPA → |
| Stomach | Glandular cells | Medium | Protein (IHC) | HPA → |
| Testis | Round or early spermatids | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced MTRR IHC Tips
Use the catalog antibody’s paraffin-section example as a starting point, then judge MTRR staining by compartment, cell type and controls.
Which retrieval conditions should I try first for MTRR in paraffin sections?
Start with heat-mediated EDTA retrieval at pH 8.0 (datasheet A01401-1). The selected paraffin-section example paired it with 10% goat-serum blocking, 2 μg/ml primary antibody overnight at 4°C, and DAB detection (datasheet A01401-1). If staining is weak, compare another buffer or heating schedule only after checking section integrity, primary-antibody handling and development controls (standard IHC practice). Keep retrieval and detection conditions identical across comparison sections, and judge improvement by cytoplasmic signal in expected cells alongside background in negative controls (UniProt Q9UBK8; HPA tissue IHC; standard IHC practice).
How should I troubleshoot weak MTRR staining when fixation histories differ?
The selected image identifies a paraffin-embedded section but does not report its fixative, so target-specific MTRR fixation sensitivity is unknown (datasheet A01401-1). Record each specimen’s fixative and fixation duration, then compare sections using the same EDTA retrieval at pH 8.0 and the same detection conditions (datasheet A01401-1; standard IHC practice). Include a previously staining section in each run to distinguish specimen variation from an assay failure (standard IHC practice). Interpret differences cautiously: altered staining across fixation histories alone cannot establish a MTRR-specific fixation effect without matched material and controls (standard IHC practice).
What staining pattern is credible for MTRR in chromogenic IHC?
Expect predominantly cytoplasmic staining when evaluating MTRR in tissue sections (UniProt Q9UBK8; HPA tissue IHC). Cytosolic localisation is supported by HPA subcellular data, while its nucleoplasmic and intermediate-filament assignments are uncertain, so isolated nuclear or filament-like DAB staining needs additional validation (HPA subcellular). MTRR has no predicted transmembrane segment, making a sharp membrane-only pattern difficult to reconcile with its annotated topology (UniProt Q9UBK8 topology). Compare signal with neighbouring cells and a no-primary control under matched development conditions before accepting an unusual compartment as biological (standard IHC practice).
Could MTRR isoforms or epitope accessibility explain inconsistent staining?
MTRR has 2 annotated isoforms, A and B, but the supplied catalog example does not define which isoforms its antibody detects (UniProt Q9UBK8; datasheet A01401-1). Its flavodoxin-like region spans residues 5–147, its FAD-binding region spans 271–533, and annotated phosphoserines occur at 171 and 189 (UniProt Q9UBK8). Without a mapped immunogen or epitope, none of those features establishes why a particular section stains weakly (standard IHC interpretation). Compare retrieval conditions and independent antibody validation before attributing a staining difference to splice form, phosphorylation or epitope masking (standard IHC practice).
How can IF help check an ambiguous MTRR IHC pattern?
Use IF as a separate validation experiment and assess whether MTRR signal overlaps the expected cell population, such as goblet cells in bronchus or glandular cells in stomach (HPA tissue IHC; standard IF practice). Multiplex with a validated marker for that cell type, and choose spectrally separated fluorophores while checking tissue autofluorescence in unstained controls (standard IF practice). Because MTRR is cytosolic and has no transmembrane segment, optimise permeabilisation for access to an intracellular epitope rather than assuming surface staining (UniProt Q9UBK8 topology; HPA subcellular; standard IF practice). Compare cellular localisation with the chromogenic result, while recognising that the selected tissue caption supplies no IF protocol (datasheet A01401-1).
How do I reduce diffuse DAB background without losing MTRR signal?
First separate diffuse background from plausible cytoplasmic MTRR staining by comparing matched no-primary controls and expected cell populations (UniProt Q9UBK8; HPA tissue IHC; standard IHC practice). The selected example used 10% goat serum before 2 μg/ml rabbit primary antibody overnight at 4°C (datasheet A01401-1). For peroxidase-based DAB detection, check endogenous-peroxidase blocking, secondary-antibody specificity, washing and chromogen development as general IHC controls (standard IHC practice). Change one variable at a time and retain a known staining section, so reduced background is not mistaken for loss of assay sensitivity (standard IHC practice).
How should I quantify MTRR staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic scoring region before measuring DAB signal, because MTRR is annotated as cytoplasmic and HPA reports cell-dependent tissue staining (UniProt Q9UBK8; HPA tissue IHC). An H-score combines the percentage of cells at each intensity; alternatively report percent positive cells or positive-cell density per mm² when those measures suit the question (standard IHC practice). Normalise to the relevant viable tissue area or eligible-cell count, and apply the same positivity threshold and imaging settings across sections (standard IHC practice). Exclude necrosis and section edges using predefined rules, then report biological replicates and scoring agreement where available (standard IHC practice).
When should an apparent MTRR-positive region be considered artefactual?
A credible result shows cellular staining consistent with MTRR’s cytoplasmic annotation and the tissue’s expected positive population, rather than an isolated edge or necrotic area (UniProt Q9UBK8; HPA tissue IHC; standard IHC practice). HPA reports high staining in bronchial goblet cells and no detected staining in adipocytes, providing cell-level reference points rather than universal controls for every specimen (HPA tissue IHC). Treat membrane-only or isolated nuclear DAB signal cautiously because MTRR lacks a transmembrane segment and HPA labels nucleoplasmic localisation uncertain (UniProt Q9UBK8 topology; HPA subcellular). Check no-primary and endogenous-peroxidase controls, tissue morphology and repeat sections before interpreting unexpected staining as altered MTRR expression (standard IHC practice).
Boster reagents
Best MTRR / Methionine synthase reductase IHC Antibodies
A01401-1 has IHC and IF images from human stomach cancer paraffin sections (catalog image captions). Human, Mouse and Rat reactivity is listed, while the images show human tissue only (catalog reactivity; catalog image captions).
Real IHC data
Anti-MTRR Antibody ®
Cat # A01401-1
A01401-1 is listed for IHC and IF with Human, Mouse and Rat reactivity (catalog applications; catalog reactivity). Its IHC and IF images each show human stomach cancer paraffin sections (catalog IHC image caption; catalog IF image caption).
Which to pick: Choose A01401-1 for chromogenic tissue IHC: its image shows a human paraffin section with EDTA pH 8.0 retrieval, 2 μg/ml primary antibody and DAB detection; the fixative is unreported (A01401-1 IHC image caption). For IF, the same rabbit antibody has a paraffin-section image, but ICC is not shown (catalog host; A01401-1 IF image caption). For cross-species work, A01401-1 lists Mouse and Rat reactivity, though its displayed IHC and IF evidence is human only (catalog reactivity; A01401-1 IHC and IF image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UBK8 (MTRR_HUMAN, Methionine synthase reductase).
- Human Protein Atlas. MTRR tissue IHC expression (reliability: Supported).
- Human Protein Atlas. MTRR subcellular location (ICC-IF): Mainly localized to the nucleoplasm and cytosol. In addition localized to the intermediate filaments..
- Human Protein Atlas. MTRR antibody validation summary (1 antibodies).
- Aberrant Expression of Folate Metabolism Enzymes and Its Diagnosis and Survival Prediction in Ovarian Carcinoma. Analytical cellular pathology (Amsterdam) 2019 — PMC6462347.
- Disruption of Folate Metabolism Causes Poor Alignment and Spacing of Mouse Conceptuses for Multiple Generations. Frontiers in cell and developmental biology 2021 — PMC8703036.
- Analysis of spermatogenesis and fertility in adult mice with a hypomorphic mutation in the Mtrr gene. Reproduction, fertility, and development 2019 — PMC7116358.
- PubMed PMID:10564814 — UniProt-cited evidence.
- PubMed PMID:9501215 — UniProt-cited evidence.
- PubMed PMID:15372022 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MTRR (Methionine synthase reductase). Expected localisation Cytoplasmic staining in tissue sections (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01401-1). Validated IHC antibody: A01401-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Goblet cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human stomach cancer tissue; fixative not specified (datasheet A01401-1); 2. Fixation: Image fixative and duration unreported (datasheet A01401-1); 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 A01401-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01401-1); 8. Primary antibody: Rabbit anti-MTRR, 2-5 μg/ml (datasheet A01401-1); 9. Primary incubation: Overnight at 4 °C (datasheet A01401-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Adipocytes may lack detectable staining (HPA tissue IHC); Isoform / epitope: 2 isoforms (A/B); epitope coverage is unknown (UniProt); Regulation: Stimulus-linked regulation is not established (UniProt). Evidence: UniProt Q9UBK8, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm and cytosol. In addition…, Datasheet A01401-1, PMC6462347, PMC8703036. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MTRR/immunohistochemistry-ihc-guide.