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MTTP / Microsomal triglyceride transfer protein large subunit · IHC design guide
Design Immunohistochemistry for MTTP
Plan MTTP staining in paraffin sections using the documented cytoplasmic pattern in intestinal glandular cells and hepatocytes (HPA tissue IHC). The guide covers antibody dilution, detection and interpretation while accounting for uncertain tissue-staining reliability (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); ER and Golgi location (UniProt) | |
| Staining pattern | Glandular cells and hepatocytes show cytoplasmic staining (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 across samples. (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target binding; staining reliability is uncertain (HPA tissue IHC) | |
| Regulation | Expression regulation is not specified (UniProt) | |
| Isoform / epitope | 2 isoforms; mature chain starts at residue 19; epitope coverage unknown (UniProt) |
Section 1
Recommended MTTP IHC & IF Protocols
The catalog antibody protocol is followed by three published MTTP IHC protocols (datasheet A01715-1; 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). Citrate pH 6.0 for 15 min is a published alternative (PMC11055869).
Section 2
What Is the Expected MTTP Staining Pattern?
MTTP is an endoplasmic reticulum protein also annotated in the Golgi, with no transmembrane segment (UniProt P55157). In paraffin-section IHC, expect cytoplasmic staining in intestinal glandular cells and hepatocytes (HPA tissue IHC). Interpret the pattern cautiously: HPA rates its tissue IHC profile Uncertain because staining and RNA data have low consistency, and presumed off-target staining was disregarded (HPA tissue IHC).
| Strong cytoplasmic staining in duodenal or small-intestinal glandular cells; moderate staining in hepatocytes. | This matches HPA's observed High glandular-cell and Medium hepatocyte staining (HPA tissue IHC). A fine intracellular pattern is compatible with ER localization (UniProt P55157); IHC alone does not prove that every stained structure is ER. |
| Dominant nuclear or cell-surface staining, with little cytoplasmic signal in expected positive cells. | This does not fit MTTP's ER/Golgi annotation or the reported cytoplasmic tissue profile (UniProt P55157; HPA tissue IHC). Check the antibody and detection controls before assigning that compartment to MTTP. |
| Strong staining in adipocytes or another cell population outside the reported positive pattern. | HPA reports adipocytes in adipose tissue as Not detected and disregarded presumed off-target staining (HPA tissue IHC). Consider cross-reactivity; signal that persists without primary antibody may reflect endogenous detection activity (standard IHC practice). |
| Broad, nearly uniform color across tissue, including extracellular spaces or expected negative cells. | This obscures the cell-specific cytoplasmic pattern reported by HPA (HPA tissue IHC). Evaluate background with a no-primary control and review blocking, washes, and chromogen development (standard IHC practice). |
| No detectable staining in duodenal or small-intestinal glandular cells on a processed positive-control section. | Those cells are reported High by HPA, so an absent signal warrants a technical check (HPA tissue IHC). Review the catalog antibody's IHC-P instructions, retrieval, dilution, and detection controls (standard IHC practice). |
💡Expected MTTP appearanceCall positive a cell-associated cytoplasmic signal, strongest in intestinal glandular cells and present at lower intensity in hepatocytes (HPA tissue IHC); dominant nuclear, surface, or diffuse field-wide color is suspect (UniProt P55157; HPA tissue IHC).
| Tissue and cell selection | Duodenal and small-intestinal glandular cells are reported High; hepatocytes are Medium (HPA tissue IHC). Include a reported positive tissue when interpreting a weak experimental section. |
| Intracellular location and processing | MTTP has a signal peptide at residues 1–18, a mature chain at 19–894, and no transmembrane segment (UniProt P55157). These annotations support an intracellular interpretation; they do not identify the antibody epitope. |
| Tissue IHC confidence | HPA labels the tissue profile Uncertain and one listed antibody's IHC validation Uncertain (HPA tissue IHC; HPA antibodies: HPA054862). Treat an isolated unexpected stain as provisional. |
| Isoforms and antibody coverage | UniProt lists 2 MTTP isoforms (UniProt P55157). The supplied record gives no epitope or isoform coverage for the catalog antibody, so it cannot explain a discrepant stain by isoform choice. |
| IF/ICC Q&A: What localization should be expected? | HPA supports ER localization in ICC-IF and calls cytosol uncertain; UniProt also annotates Golgi (HPA subcellular; UniProt P55157). Use the separate IF/ICC guide for that application. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control intestinal glandular cells are blank. | The result conflicts with reported High staining, but HPA's tissue profile is Uncertain (HPA tissue IHC). A failed IHC step remains possible (standard IHC practice). | Confirm the catalog antibody's IHC-P procedure, then check retrieval, primary dilution, and detection with appropriate run controls (standard IHC practice). |
| Hepatocytes stain weakly beside strong intestinal glandular cells. | HPA reports Medium hepatocyte staining versus High intestinal glandular-cell staining (HPA tissue IHC). | Score the two cell types against their respective reported levels; keep exposure and chromogen development comparable across sections (standard IHC practice). |
| Adipocytes show strong staining. | HPA reports adipose adipocytes as Not detected, and its overall tissue profile is Uncertain (HPA tissue IHC). Cross-reactivity or detection background is plausible (standard IHC practice). | Compare a no-primary control and a known-positive section in the same run before interpreting the adipocyte signal (standard IHC practice). |
| Color appears predominantly nuclear or along plasma membranes. | That distribution conflicts with ER/Golgi annotation and HPA's cytoplasmic tissue profile (UniProt P55157; HPA tissue IHC). | Recheck morphology and counterstain, then compare primary-free and positive-control sections; do not score the unexpected compartment as MTTP without further validation (standard IHC practice). |
| Most cells and empty spaces acquire diffuse chromogen. | Non-cell-specific color prevents assessment of HPA's reported cell-specific pattern (HPA tissue IHC). Excess background can arise during detection (standard IHC practice). | Inspect the no-primary control; review blocking, wash stringency, and chromogen development using standard IHC controls (standard IHC practice). |
| The stain differs from an ICC-IF image. | HPA's ICC-IF record supports ER localization but marks cytosol uncertain; its tissue IHC reliability is also Uncertain (HPA subcellular; HPA tissue IHC). | Compare each result within its own application and controls. For paraffin IHC, prioritize the observed tissue and cell pattern while retaining HPA's reliability caveat (HPA tissue IHC). |
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; its glandular cells should stain strongly (HPA: High in duodenal glandular cells). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes), and compare nonglandular cells on the duodenal slide with the no-primary control as an internal background reference rather than assuming they lack MTTP.
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 no-primary (secondary-only) and concentration-matched rabbit IgG controls (caption: rabbit primary; standard IHC practice), plus an MTTP knockout specimen or validated peptide-block control (standard IHC practice). Check endogenous peroxidase and endogenous biotin in duodenum because the documented detection uses biotinylated secondary antibody and SABC with DAB (caption; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A01715-1 paraffin-section caption does not state the fixative (caption). The documented IHC procedure uses heat-mediated retrieval in EDTA at pH 8.0, but the evidence does not establish whether retrieval is required or whether frozen sections or IF are easier (caption). Endogenous biotin can complicate the documented SABC detection in duodenum, so assess background with the technical controls (caption; 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
Troubleshoot MTTP staining in paraffin sections by checking retrieval, compartment, cell type, controls, and scoring before interpreting differences in signal.
Which retrieval condition should I start with for paraffin section MTTP staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin section MTTP IHC (datasheet A01715-1). The selected tissue image used that condition before incubation with 2 μg/mL antibody overnight at 4°C, so reproduce its retrieval and antibody conditions when establishing a baseline (caption A01715-1). Compare a retrieved section with a section processed without retrieval to judge whether heating improves the expected cellular pattern (standard IHC practice; UniProt P55157 localisation). Keep section thickness, heating time, detection, and imaging conditions consistent across comparisons so a change in staining can be attributed to retrieval (standard IHC practice).
How should I investigate weak MTTP staining when the tissue fixative is unknown?
The selected image identifies a paraffin-embedded section but does not state its fixative, and target-specific MTTP fixation sensitivity is unknown (caption A01715-1). Record the fixative, fixation duration, processing history, and section age for each specimen before comparing staining intensity (standard IHC practice). Run the same IHC-validated antibody and EDTA pH 8.0 retrieval on a known positive control alongside the affected section (datasheet A01715-1; HPA: cytoplasmic expression in liver and small intestine). If the control stains and the specimen does not, review preservation and tissue quality while keeping the antibody and detection conditions matched (standard IHC practice).
What subcellular pattern should count as plausible MTTP staining in IHC?
Expect a predominantly cytoplasmic pattern in appropriate cells, consistent with the tissue IHC profile and MTTP localisation to the endoplasmic reticulum and Golgi apparatus (HPA tissue IHC profile; UniProt P55157 localisation). MTTP has a signal peptide at residues 1–18 and no transmembrane segment, so do not require a continuous plasma membrane outline when scoring positivity (UniProt P55157 topology and processing). Compare staining within glandular cells or hepatocytes with neighbouring structures on the same section (HPA: high in duodenal and small-intestinal glandular cells; medium in hepatocytes). Predominantly nuclear or sharply membrane-restricted signal warrants review of morphology, controls, and detection background before it is assigned to MTTP (UniProt P55157 localisation; standard IHC practice).
Can a negative stain exclude both MTTP isoforms?
A negative section cannot exclude both MTTP isoforms without evidence that the antibody recognizes an epitope shared by isoforms 1 and 2 (UniProt P55157 isoforms; standard IHC interpretation). Check the catalog antibody’s stated immunogen or mapped epitope against each isoform sequence before interpreting discordant specimens (standard IHC practice). The record identifies a signal peptide at residues 1–18 and a mature chain spanning residues 19–894, making epitope position relevant to that comparison (UniProt P55157 processing). If epitope mapping is unavailable, describe the result as staining with this antibody under the tested retrieval conditions and corroborate uncertain cases with an independent antibody or method (standard IHC practice).
How can I examine MTTP by IF while checking cell identity and background?
For a separate IF experiment, multiplex MTTP with a marker identifying the expected glandular cells or hepatocytes, and assess whether signal falls within those marked cells (HPA: high in intestinal glandular cells; medium in hepatocytes; standard IF practice). Choose fluorophores after checking unstained tissue in each channel, placing the weaker signal where tissue autofluorescence is lowest (standard IF practice). MTTP is associated with the endoplasmic reticulum and Golgi apparatus and has no transmembrane segment, so include permeabilisation sufficient for antibody access to its intracellular epitope (UniProt P55157 localisation and topology; standard IF practice). Validate permeabilisation and antibody specificity with matched controls before comparing IF images with chromogenic IHC patterns (standard IF practice).
How do I distinguish diffuse MTTP staining from chromogenic background?
First compare the stained section with a matched section lacking primary antibody to identify signal from secondary reagents or the detection system (standard IHC practice). The selected image used a biotinylated secondary antibody, a streptavidin–biotin complex, and DAB, so evaluate background at each of those steps when reproducing that workflow (caption A01715-1; standard IHC practice). Apply a peroxidase block for chromogenic detection and inspect tissue edges, damaged areas, and pigment before calling diffuse brown signal positive (standard IHC practice). Give greater weight to staining in the expected cells and cytoplasmic compartment than to colour distributed indiscriminately across tissue structures (HPA tissue IHC profile; UniProt P55157 localisation; standard IHC interpretation).
What should I score when comparing MTTP across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and anatomical region before scoring, because expected MTTP expression differs between intestinal glandular cells and hepatocytes (HPA: high in duodenal and small-intestinal glandular cells; medium in hepatocytes). For a cellular readout, record the percentage of positive target cells and staining intensity, then calculate an H-score using the same thresholds across sections (standard IHC practice). Alternatively, report positive cells per mm² of viable target tissue when cell counts are more reliable than intensity (standard IHC practice). Normalise to the number or area of evaluable target cells, exclude necrotic and damaged regions, and keep detection and image settings consistent (standard IHC practice).
Which findings would make an apparent MTTP positive result questionable?
A credible result should favour the expected cytoplasmic compartment and relevant cell types, with a staining pattern that survives comparison with controls (UniProt P55157 localisation; HPA tissue IHC profile; standard IHC interpretation). Question predominantly nuclear staining, strong signal in an unexpected cell population, or colour concentrated at section edges or necrotic areas (UniProt P55157 localisation; standard IHC practice). Check a section without primary antibody and review the peroxidase block if DAB colour suggests endogenous enzyme activity (standard IHC practice). Treat an isolated positive pattern cautiously: the HPA tissue IHC assessment is uncertain because staining and RNA expression showed low consistency and presumed off-target binding was disregarded (HPA tissue IHC reliability).
Boster reagents
Best MTTP / Microsomal triglyceride transfer protein large subunit IHC Antibodies
A01715-1 has IHC images from human liver cancer and rat liver paraffin sections, plus IF data from HEPG2 cells (catalog image captions).
Real IHC data
Anti-MTTP Antibody ®
Cat # A01715-1
A01715-1 will render with its human liver cancer paraffin-section IHC figure; a second catalog caption documents rat liver paraffin-section IHC (catalog IHC image captions). The same SKU lists IF and ICC applications and has an IF image from HEPG2 cells (catalog applications and IF image caption).
Which to pick: Choose A01715-1 for human or rat paraffin-section IHC: its captions document EDTA heat retrieval at pH 8.0 and 2 μg/ml antibody; the fixative is unreported (catalog IHC image captions). For IF/ICC, A01715-1 has a HEPG2 cell IF image using 5 μg/ml antibody (catalog IF image caption). For cross-species planning, the catalog lists Human, Monkey and Rat reactivity, while the supplied IHC images document Human and Rat only; no clone type is supplied (catalog reactivity, IHC image captions and clone field).
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); ER and Golgi location (UniProt). 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 across samples. (standard IHC practice; not target-specific); Caveat: Presumed off-target binding; staining reliability is uncertain (HPA tissue IHC); Isoform / epitope: 2 isoforms; mature chain starts at residue 19 ; epitope coverage unknown (UniProt); Regulation: Expression regulation is not specified (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.