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MTTP / Microsomal triglyceride transfer protein large subunit · IHC design guide
Design Immunohistochemistry for MTTP
Plan MTTP IHC in small-intestinal glandular cells and hepatocytes, where HPA reports cytoplasmic staining (HPA tissue IHC). Use the catalog antibody’s paraffin-section protocol and interpret staining with HPA’s uncertain reliability in mind (datasheet A01715-1; HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MTTP 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 staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic in intestinal glandular cells and hepatocytes (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01715-1) | |
| Positive control | Duodenum+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 paraffin-section fixation consistent. (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target staining; reliability uncertain (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unreported (UniProt) |
Section 1
Recommended MTTP IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by three published MTTP IHC protocols for chicken ovarian tissue, paired tumor samples, and additional MTTP-stained samples (PMC11951013; PMC9534973; PMC11055869).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A01715-1) |
| Fixation | Image fixative and duration unreported (datasheet A01715-1); 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 A01715-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01715-1) |
| Primary antibody | Rabbit anti-MTTP, 2-5μg/ml (datasheet A01715-1) |
| Primary incubation | Overnight at 4 °C (datasheet A01715-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01715-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MTTP-positive staining in glandular cells of duodenum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in liver and small intestine. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A01715-1). A published alternative uses citrate pH 6.0 for 15 min (PMC11055869).
Section 2
What Is the Expected MTTP Staining Pattern?
MTTP staining should appear mainly in the cytoplasm of duodenal and small-intestinal glandular cells, with a weaker signal in hepatocytes (HPA tissue IHC: high in intestinal glandular cells; medium in hepatocytes). Its supported endoplasmic-reticulum location gives a more specific localisation expectation than cytoplasm alone (HPA ICC-IF: ER supported; UniProt P55157: ER and Golgi). Interpret chromogenic IHC cautiously: HPA rates the tissue pattern uncertain and reports presumed off-target staining (HPA tissue IHC: uncertain).
| Cytoplasmic staining in intestinal glandular cells and hepatocytes, stronger in the intestine. | This fits the observed tissue pattern (HPA tissue IHC: high in duodenum and small intestine; medium in liver). Score the relevant cells and their cytoplasmic signal; HPA's uncertain reliability means the pattern alone does not prove specificity (HPA tissue IHC: uncertain). |
| Predominantly nuclear or sharply plasma-membrane staining in otherwise positive cells. | This conflicts with MTTP's reported intracellular locations and lack of a transmembrane segment (UniProt P55157: ER, Golgi, no transmembrane segment). Consider artefact or nonspecific staining; a cytoplasmic signal in the same section does not validate the discordant compartment. |
| Strong staining in adipocytes, marrow cells, or other cells reported as undetected. | These are discrepancies with the sampled HPA tissue observations (HPA tissue IHC: adipocytes and marrow hematopoietic cells not detected). Assess cross-reactivity or endogenous chromogen-generating activity before calling them MTTP-positive; an HPA 'not detected' result is not a universal absence claim. |
| Diffuse chromogen across tissue and empty areas, obscuring cell boundaries. | Treat this as background until the assay controls identify its source (general IHC practice). Distribution that does not track the expected glandular cells or hepatocytes is weak evidence for MTTP (HPA tissue IHC: cytoplasmic expression in liver and small intestine). |
| No detectable stain in duodenal or small-intestinal glandular cells. | A negative result in a reported high-staining tissue warrants a run-level check (HPA tissue IHC: high in duodenum and small intestine). Examine tissue preservation, retrieval, antibody dilution, detection, and controls using general IHC practice before interpreting the sample as biologically negative. |
💡Expected MTTP appearanceCall a result pattern-consistent when intestinal glandular cells show strong cytoplasmic staining and hepatocytes show a lesser cytoplasmic signal; isolated nuclear, membrane, or broadly misplaced staining warrants scrutiny (HPA tissue IHC: high intestine, medium liver, uncertain reliability; UniProt P55157: ER and Golgi).
| Tissue and cell selection | Duodenum or small intestine provides an observed high-staining reference; liver offers a medium-staining comparison (HPA tissue IHC). Several sampled tissues were undetected, so their lack of signal alone is a poor test of assay sensitivity (HPA tissue IHC: not detected in listed negative tissues). |
| Intracellular location and processing | MTTP has a signal peptide at residues 1–18 and a mature chain at 19–894, without a transmembrane segment (UniProt P55157). Interpret staining against its ER and Golgi locations; the supplied record does not locate the catalog antibody's epitope (UniProt P55157: topology and processing). |
| Antibody validation and tissue reliability | HPA marks the available tissue IHC antibody HPA054862 uncertain and describes low consistency with RNA plus presumed off-target binding (HPA antibodies: IHC uncertain; HPA tissue IHC: uncertain). A matching distribution raises confidence in interpretation but does not independently establish target specificity. |
| ICC-IF comparison | Q: What can IF add? A: An ER-like pattern can support the localisation interpretation (HPA ICC-IF: ER supported). Cytosolic staining has uncertain support, and this IHC section does not establish an IF protocol or transfer IHC validation to IF (HPA ICC-IF: cytosol uncertain; HPA antibodies: application-specific status). |
| Antigen retrieval and assay settings | Use the catalog antibody's IHC-P directions and tissue controls to assess retrieval and dilution (general IHC practice). The supplied HPA and UniProt records give no MTTP-specific fixation or retrieval response, so a change in signal cannot be attributed to a documented MTTP sensitivity. |
| Situation | Likely cause | Next action |
|---|---|---|
| Intestinal reference tissue is blank. | The high-staining cell population may be absent from the section, or an assay step may have failed (HPA tissue IHC: high in intestinal glandular cells; general IHC practice). | Confirm glandular cells on the counterstain; check the run's positive control, then review the catalog IHC-P retrieval, dilution, and detection settings (general IHC practice). |
| Liver is faint while intestinal glands stain clearly. | This difference is compatible with reported staining levels (HPA tissue IHC: hepatocytes medium; intestinal glandular cells high). | Compare the same cell types and scoring threshold across sections; retain the intestinal reference when judging assay performance (general IHC practice; HPA tissue IHC: high intestine). |
| Nuclei or cell borders dominate the stain. | The compartment disagrees with the reported ER and Golgi localisation and absence of a transmembrane segment (UniProt P55157). | Review counterstain and tissue morphology, then compare omission and other assay controls for nonspecific deposition (general IHC practice). Do not count the discordant compartment as pattern-confirming MTTP. |
| Reported negative cell populations stain strongly. | Cross-reactivity or endogenous detection activity is plausible, especially given the uncertain tissue IHC assessment (HPA tissue IHC: presumed off-target binding; general IHC practice). | Check a no-primary control for detection background and compare the suspect population with the listed HPA-negative examples (general IHC practice; HPA tissue IHC: listed negatives). |
| Chromogen appears widespread or obscures the expected cells. | Background may reflect blocking, detection, or antibody concentration settings (general IHC practice); the broad distribution does not match HPA's reported tissue profile (HPA tissue IHC). | Inspect no-primary and positive controls; adjust the relevant blocking, wash, or dilution step according to the catalog IHC-P workflow (general IHC practice). |
| IHC is ambiguous, and an IF comparison is being considered. | HPA supports ER localisation by ICC-IF but calls the cytosolic component uncertain (HPA ICC-IF: ER supported; cytosol uncertain). | Use the ER observation only as a localisation cross-check, and assess IF with its own application validation (HPA ICC-IF; HPA antibodies: application-specific status). |
Experiment evidence
IHC & IF images showing MTTP staining
A01715-1 · IHCIHC analysis of MTTP using anti-MTTP antibody (A01715-1). MTTP was detected in paraffin-embedded section of human liver cancer tissue. Heat
A01715-1 · IHCIHC analysis of MTTP using anti-MTTP antibody (A01715-1). MTTP was detected in paraffin-embedded section of rat liver tissue. Heat mediated
A01715-1 · IFIF analysis of MTTP using anti-MTTP antibody (A01715-1). MTTP was detected in immunocytochemical section of HEPG2 cells. Enzyme antigen retr
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000138823-MTTP/tissue/cerebral+cortex#imid_15753635
HPA · Small intestineHuman Protein Atlas IHC, Small intestine. Source: https://www.proteinatlas.org/ENSG00000138823-MTTP/tissue/small+intestine#imid_15753528
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000138823-MTTP/tissue/colon#imid_15753569
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000138823-MTTP/tissue/liver#imid_15753554
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA051170; green = MTTP, blue = nucleus, red = microtubules.
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA051170; green = MTTP, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MTTP IHC & IF
🧪Run duodenum first: glandular cells should stain (HPA: High in duodenum glandular cells). Use adipose tissue as the negative comparator (HPA: Not detected in adipocytes); on the duodenum slide, assess nonglandular cells as an internal background comparator rather than assuming they are MTTP-negative (HPA: glandular-cell result only).
Positive control tissue: Duodenum (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MTTP in Hep-G2, SH-SY5Y, U2OS, ASC52telo, Sperm, with annotated localisation: Endoplasmic reticulum (supported), Cytosol (uncertain) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and host-matched rabbit control IgG (caption: rabbit primary antibody); use MTTP-knockout material as a biological specificity control where available (standard IHC practice). Quench endogenous peroxidase and check for endogenous biotin before using the biotin-based DAB detection shown in the caption, particularly in duodenal tissue (caption: biotinylated secondary and SABC–DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A01715-1 tissue-IHC caption does not state its fixative (caption: fixative unreported). Heat retrieval in EDTA at pH 8.0 was used for the pictured paraffin section, but retrieval dependence was not established (caption: EDTA heat retrieval). The supplied evidence does not establish that frozen sections or IF are easier; for duodenal IHC, assess endogenous biotin background with the stated SABC detection method (caption: SABC–DAB; standard IHC practice).
HPA tissue IHC evidence for MTTP
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). 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 |
|---|---|---|---|---|
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Small intestine | Glandular cells | High | Protein (IHC) | HPA → |
| Liver | Hepatocytes | 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 MTTP IHC Tips
Use compartment, cell type and matched controls to evaluate MTTP staining in paraffin sections; the available tissue IHC reliability assessment is uncertain (HPA tissue IHC).
Which retrieval condition should I start with for MTTP in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A01715-1). The selected tissue-IHC caption used that condition before an overnight primary incubation at 4°C, but did not report a heating duration (caption A01715-1). If staining is weak, compare controlled changes in heating duration on adjacent sections while keeping antibody concentration and detection constant (standard IHC practice). Include a liver or small-intestine section with the test batch, and judge improvement by cytoplasmic staining in expected cells rather than stronger staining everywhere (HPA tissue IHC; standard IHC practice).
How should I assess fixation effects on MTTP staining?
MTTP-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state its fixative (caption A01715-1). Record fixative, fixation duration, tissue thickness and processing history for each specimen, then compare similarly processed sections before attributing intensity differences to biology (standard IHC practice). Keep retrieval at EDTA pH 8.0 and the detection method consistent during that comparison (datasheet A01715-1; standard IHC practice). If a new fixation condition produces weak staining, check an expected positive tissue in the same run and optimize retrieval separately; tissue expression patterns do not establish a fixation effect (HPA tissue IHC; standard IHC practice).
What staining pattern is plausible for MTTP in tissue sections?
Expect intracellular cytoplasmic staining in hepatocytes and small-intestinal glandular cells, with potentially uneven cytoplasmic distribution reflecting the endoplasmic reticulum (HPA tissue IHC; UniProt P55157 localisation). MTTP is reported in the endoplasmic reticulum and Golgi, and colocalizes with P4HB/PDI in the endoplasmic reticulum (UniProt P55157 localisation). A crisp nuclear or cell-surface-only DAB pattern is inconsistent with those locations and warrants review of morphology, counterstain and controls (UniProt P55157 localisation; standard IHC practice). Treat diffuse cytoplasmic staining cautiously because the HPA tissue-IHC assessment is uncertain and reports presumed off-target binding (HPA tissue IHC).
Can this staining distinguish MTTP isoforms or a processed protein?
Do not assign staining to a particular isoform without an antibody epitope map: the record lists 2 MTTP isoforms, but no epitope for this antibody is supplied (UniProt P55157 isoforms; datasheet A01715-1). MTTP has a signal peptide at residues 1–18 and a mature chain at 19–894, so recognition of either form depends on where the antibody binds (UniProt P55157 processing). The record lists no transmembrane segment or glycosylation sites; neither fact identifies the recognized epitope (UniProt P55157 topology and glycosylation). For a claimed isoform difference, obtain sequence-specific epitope information and compare appropriately validated reagents on matched sections (standard IHC practice).
How should I adapt this MTTP question to a multiplex IF experiment?
For IF, pair MTTP with a validated hepatocyte or intestinal glandular-cell marker and, if useful, an ER marker such as P4HB/PDI; expected tissue expression and ER colocalization support those comparisons (HPA tissue IHC; UniProt P55157 localisation). Select spectrally separated fluorophores and place the weaker signal in a channel with low measured tissue autofluorescence (standard IF practice). Permeabilize sufficiently to expose intracellular MTTP, then compare mild and stronger conditions because the antibody epitope is unreported and MTTP has no transmembrane segment (UniProt P55157 topology; standard IF practice). Include single-label, secondary-only and unstained controls before interpreting overlap (standard IF practice).
What should I check when MTTP DAB staining is widespread?
First compare the primary-omitted control with the test section to identify detection-system background, and inspect blank tissue regions for precipitated chromogen (standard IHC practice). The selected caption used a biotinylated secondary, streptavidin-biotin complex and DAB, so assess endogenous biotin and block endogenous peroxidase as appropriate for that workflow (caption A01715-1; standard IHC practice). Its 10% goat-serum block and 2 µg/mL primary concentration provide documented starting conditions, not proof that every tissue will have low background (caption A01715-1). Broad staining outside expected cells deserves particular scrutiny because presumed off-target binding contributed to an uncertain HPA reliability rating (HPA tissue IHC).
How can I score MTTP staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population before scoring: hepatocytes in liver or glandular cells in small intestine are the documented positive populations (HPA tissue IHC). For chromogenic sections, report the percentage of positive cells and an H-score from 0–300, calculated from percentages at intensity grades 0–3 (standard IHC practice). Normalize comparisons to the number of eligible cells or viable tissue area, and apply the same thresholds, acquisition settings and exclusion rules across sections (standard IHC practice). Score edge damage, necrosis and nonspecific deposits separately; the uncertain HPA reliability assessment makes control-supported thresholds especially useful (HPA tissue IHC; standard IHC practice).
When should an apparent MTTP-positive section be questioned?
A plausible positive is cytoplasmic staining in hepatocytes or intestinal glandular cells, consistent with reported tissue expression and ER/Golgi localization (HPA tissue IHC; UniProt P55157 localisation). Question dominant nuclear or cell-surface staining, or strong signal confined to cells outside the expected population, and compare it with controls before assigning MTTP expression (UniProt P55157 localisation; standard IHC practice). Edge effects, necrotic regions, endogenous peroxidase and endogenous biotin can mimic or amplify DAB signal in the selected detection workflow (caption A01715-1; standard IHC practice). Independent confirmation matters because HPA rates its tissue-IHC evidence uncertain, citing low staining–RNA consistency and presumed off-target binding (HPA tissue IHC).
Boster reagents
Best MTTP / Microsomal triglyceride transfer protein large subunit IHC Antibodies
The IHC-validated anti-MTTP antibody has paraffin-section images from human liver cancer and rat liver, plus IF data from HEPG2 cells (A01715-1 image captions).
Real IHC data
Anti-MTTP Antibody ®
Cat # A01715-1
A01715-1 will render with its IHC figure from a paraffin-embedded human liver cancer section; a second catalog caption documents rat liver IHC (A01715-1 IHC captions). The same SKU lists IF and ICC applications and has an IF image from HEPG2 cells (catalog applications; A01715-1 IF caption).
Which to pick: Choose A01715-1 for chromogenic paraffin-section IHC in human or rat tissue: its own captions document EDTA pH 8 retrieval and DAB detection; the fixative is unreported (A01715-1 IHC captions). For IF/ICC, the same SKU lists both applications and shows IF in HEPG2 cells at 5 μg/mL (catalog applications; A01715-1 IF caption). For cross-species planning, human and rat have IHC captions; Monkey is listed as reactive without an IHC figure, and clonality is unreported (A01715-1 IHC captions; catalog reactivity; catalog clone).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P55157 (MTP_HUMAN, Microsomal triglyceride transfer protein large subunit).
- Human Protein Atlas. MTTP tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. MTTP subcellular location (ICC-IF): Localized to the endoplasmic reticulum and cytosol..
- Human Protein Atlas. MTTP antibody validation summary (2 antibodies).
- Ovarian expression of functional MTTP and apoB for VLDL assembly and secretion in chickens. Poultry science 2025 — PMC11951013.
- Adipocyte-Derived Exosomal MTTP Suppresses Ferroptosis and Promotes Chemoresistance in Colorectal Cancer. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2022 — PMC9534973.
- A MTA2-SATB2 chromatin complex restrains colonic plasticity toward small intestine by retaining HNF4A at colonic chromatin. Nature communications 2024 — PMC11055869.
- Epigenome-guided analysis of the transcriptome of plaque macrophages during atherosclerosis regression reveals activation of the Wnt signaling pathway. PLoS genetics 2014 — PMC4256277.
- PubMed PMID:8111381 — UniProt-cited evidence.
- PubMed PMID:8361539 — UniProt-cited evidence.
- PubMed PMID:7545943 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MTTP (Microsomal triglyceride transfer protein large subunit). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01715-1). Validated IHC antibody: A01715-1 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Duodenum (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A01715-1); 2. Fixation: Image fixative and duration unreported (datasheet A01715-1); 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 A01715-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01715-1); 8. Primary antibody: Rabbit anti-MTTP, 2-5μg/ml (datasheet A01715-1); 9. Primary incubation: Overnight at 4 °C (datasheet A01715-1). Watch for: Fixation: Keep paraffin-section fixation consistent. (standard IHC practice; not target-specific); Caveat: Presumed off-target staining; reliability uncertain (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences unreported (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt P55157, HPA tissue IHC · Uncertain, HPA ICC-IF · endoplasmic reticulum and cytosol., Datasheet A01715-1, PMC11951013, PMC9534973. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MTTP/immunohistochemistry-ihc-guide.