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DGAT1 / Diacylglycerol O-acyltransferase 1 · IHC design guide
Design Immunohistochemistry for DGAT1
Plan chromogenic DGAT1 IHC on paraffin sections using the catalog antibody at 1:50 (datasheet M02522). Compare cytoplasmic staining in intestinal endocrine cells with adipocytes reported as undetected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DGAT1 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 | General cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic; high in intestinal endocrine cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M02522) | |
| Positive control | Adrenal gland+4 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); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M02522) | |
| Caveat | Adipocytes were not detected despite intestinal staining (HPA tissue IHC) | |
| Regulation | No staining-linked regulator reported (UniProt) | |
| Isoform / epitope | No annotated isoforms; 9 spans make epitope side relevant (UniProt) |
Section 1
Recommended DGAT1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA pH 8.0 retrieval (datasheet M02522). The published protocols below cover patient biopsies and rat brain tissue (PMC6058035; PMC12082899).
| Sample | Paraffin-embedded human stomach cancer tissue; fixative not specified (datasheet M02522) |
| Fixation | Image fixative and duration unreported (datasheet M02522); 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 M02522); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M02522) |
| Primary antibody | Rabbit monoclonal (clone 23D11) anti-DGAT1, 1:50 (datasheet M02522) |
| Primary incubation | Overnight at 4 °C (datasheet M02522) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M02522) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DGAT1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression, most abundant in small intestine and duodenum. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet M02522). For rat brain optimization, the published alternatives are EDTA pH 9.0 and citrate pH 6.0 (PMC12082899).
Section 2
What Is the Expected DGAT1 Staining Pattern?
DGAT1 is an endoplasmic reticulum membrane protein with 9 transmembrane segments (UniProt O75907 topology). In paraffin section IHC, expect predominantly cytoplasmic staining, especially in intestinal cells (HPA tissue IHC: general cytoplasmic expression, most abundant in small intestine and duodenum). HPA rates the tissue staining pattern “Enhanced” for agreement with RNA expression, with external verification pending (HPA tissue IHC: reliability).
| Cytoplasmic staining in duodenal or small intestinal endocrine cells. | This fits an expected positive result: HPA reports High staining in these cells and general cytoplasmic expression (HPA tissue IHC). Record the stained cell type and intensity when comparing sections (general IHC practice). |
| Staining appears chiefly nuclear or extracellular in an IHC section. | Treat the compartment as suspect and check controls, since the established tissue pattern is cytoplasmic (HPA tissue IHC) and DGAT1 resides in the endoplasmic reticulum membrane (UniProt O75907). Nucleolar staining has been reported by ICC-IF, so its relevance to paraffin IHC needs separate validation (HPA subcellular). |
| Strong staining appears in cells recorded as undetected, such as adipocytes or lymph node germinal center cells. | This conflicts with those cell-specific HPA observations (HPA tissue IHC: Not detected). Consider antibody cross-reactivity or endogenous chromogenic detection activity, and compare with an appropriate negative control (general IHC practice); a single discrepant section does not establish the cause. |
| Color covers most structures without a clear cell or compartment pattern. | Diffuse background cannot establish DGAT1 localization. Review the no-primary control, blocking, washes and detection conditions (general IHC practice), then judge whether cytoplasmic staining is distinguishable from background (HPA tissue IHC: general cytoplasmic expression). |
| No staining appears in duodenal or small intestinal endocrine cells. | Those are High staining cell populations in the HPA record (HPA tissue IHC). If the expected cells are present, check section quality and the IHC detection workflow before interpreting the result as DGAT1 absence (general IHC practice). |
💡Expected DGAT1 appearanceCall positive a discernible, predominantly cytoplasmic signal in intestinal endocrine cells with High staining expected in duodenum or small intestine; broad background or chiefly nuclear color is suspect (HPA tissue IHC: High; general cytoplasmic expression; UniProt O75907: endoplasmic reticulum membrane).
| Membrane topology (UniProt O75907) | DGAT1 spans the membrane 9 times, with cytoplasmic and lumenal regions (UniProt O75907 topology). Epitope location for the listed antibody is not supplied, so topology alone cannot predict which retrieval or permeabilisation setting will work. |
| Tissue and cell choice (HPA tissue IHC) | HPA reports High staining in duodenal and small intestinal endocrine cells and rectal enterocytes (HPA tissue IHC). It reports adipocytes as Not detected; evaluate named cell populations rather than assigning one score to an entire organ. |
| Antibody evidence (HPA antibodies; HPA tissue IHC) | CAB032853 has an Enhanced IHC status (HPA antibodies). HPA describes tissue reliability as Enhanced through staining and RNA agreement, with external verification pending (HPA tissue IHC); neither designation validates every new specimen or staining run. |
| IF/ICC: what pattern should be expected? (HPA subcellular) | HPA places the main ICC-IF signal in the endoplasmic reticulum and also reports nucleoli and nucleoli rim (HPA subcellular: approved locations). These ICC-IF observations do not establish a paraffin IHC nuclear pattern; IF/ICC methods belong in its separate guide. |
| Protein processing (UniProt O75907) | UniProt records one chain spanning residues 1–488, with no signal peptide or propeptide (UniProt O75907 processing). There is no supplied processing or shedding evidence that would explain an extracellular IHC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive intestinal cells show no signal (HPA tissue IHC: High). | Possible section, retrieval, primary-antibody or detection failure; the supplied sources do not identify DGAT1-specific fixation sensitivity (general IHC practice). | Confirm the expected cells are present, then review the validated IHC workflow and a run control before changing one condition at a time (general IHC practice). |
| Diffuse color obscures the cytoplasm. | Nonspecific binding, insufficient washing or endogenous detection activity can raise chromogenic background (general IHC practice). | Inspect a no-primary control; review blocking, washes and the detection system, then reassess the cytoplasmic pattern (general IHC practice; HPA tissue IHC: cytoplasmic expression). |
| Strong nuclear staining dominates paraffin IHC. | The dominant tissue pattern conflicts with HPA's general cytoplasmic observation (HPA tissue IHC). ICC-IF nucleolar localization does not by itself explain the IHC result (HPA subcellular). | Compare negative and positive controls and repeat the compartment assessment; interpret a nuclear IHC result cautiously until independently supported (general IHC practice). |
| Strong signal appears in HPA-undetected cells. | For adipocytes or lymph node germinal center cells, strong staining conflicts with the reported Not detected level (HPA tissue IHC); cross-reactivity or endogenous detection activity is possible (general IHC practice). | Verify cell identity and review the no-primary control before assigning the color to DGAT1 (general IHC practice). |
| An entire intestinal section is scored uniformly positive. | Whole-section scoring can hide cell-specific variation: HPA identifies High endocrine-cell staining in duodenum and small intestine (HPA tissue IHC). | Score recognizable cell populations separately and record their intensity and compartment (general IHC practice). |
| ICC-IF nucleolar signal is used to validate nuclear IHC. | HPA reports nucleoli and nucleoli rim as additional ICC-IF locations, while its tissue IHC profile is generally cytoplasmic (HPA subcellular; HPA tissue IHC). | Keep the assay-specific observations separate; use the tissue IHC pattern and IHC controls to interpret the paraffin section (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing DGAT1 staining
M02522 · IHCIHC analysis of DGAT1 using anti-DGAT1 antibody (M02522). DGAT1 was detected in a paraffin-embedded section of human stomach cancer tissue.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000185000-DGAT1/tissue/cerebral+cortex#imid_8868796
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000185000-DGAT1/tissue/colon#imid_8868714
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000185000-DGAT1/tissue/liver#imid_8868723
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000185000-DGAT1/tissue/kidney#imid_8868716
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB032853; green = DGAT1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB032853; green = DGAT1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DGAT1 IHC & IF
🧪Run small intestine first: endocrine cells should stain strongly (HPA: High in small intestine endocrine cells). Use adipose tissue as the negative tissue, focusing on adipocytes (HPA: Not detected in adipocytes); on the positive slide, compare endocrine cells with neighboring cells that lack specific signal, without assuming every other cell type is negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show DGAT1 in A-431, U-251MG, U2OS, with annotated localisation: Endoplasmic reticulum (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a matched rabbit IgG isotype control, matching clonality where applicable (selected-SKU caption: rabbit primary antibody). Use DGAT1 knockout material as a biological specificity control; for chromogenic detection in small intestine, quench endogenous peroxidase and inspect the no-primary slide for background (selected-SKU caption: HRP/DAB detection).
⚠️Feasibility: A DGAT1-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected-SKU tissue-IHC caption does not state the fixative. Heat retrieval in EDTA at pH 8.0 is a documented starting condition for paraffin sections, but its necessity has not been established (selected-SKU caption: EDTA retrieval, pH 8.0); the evidence does not show that frozen sections or IF are easier. Check intestinal sections for endogenous peroxidase background under HRP/DAB detection (selected-SKU caption: HRP/DAB detection).
HPA tissue IHC evidence for DGAT1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | High | Protein (IHC) | HPA → |
| Rectum | Enterocytes | High | Protein (IHC) | HPA → |
| Small intestine | Endocrine cells | High | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced DGAT1 IHC Tips
Troubleshoot DGAT1 staining in paraffin sections by assessing retrieval, cell identity and subcellular pattern against the supplied IHC evidence (datasheet M02522; HPA tissue IHC).
How should I troubleshoot weak DGAT1 staining after antigen retrieval?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections stained with M02522 (datasheet M02522). The selected tissue image used this retrieval before 1:50 primary incubation overnight at 4°C, so reproduce those linked conditions when investigating a weak signal (datasheet M02522). Because DGAT1 crosses the endoplasmic reticulum membrane 9 times, retrieval may expose or damage different epitopes unevenly; the antibody epitope is unspecified here (UniProt O75907 topology). If staining remains weak, compare a controlled alternative retrieval condition on adjacent sections while keeping antibody concentration, detection and tissue processing constant; assess signal in expected intestinal compartments against matched negative controls (HPA tissue IHC).
Could fixation explain inconsistent DGAT1 staining between paraffin specimens?
Target-specific sensitivity of DGAT1 staining to fixation is unknown from the supplied IHC evidence; the M02522 image identifies a paraffin section but does not state its fixative (datasheet M02522). Record the actual fixative, fixation duration and time before processing for each specimen, then compare sections with known handling histories using identical retrieval and detection settings (standard IHC practice). Begin the M02522 comparison with EDTA at pH 8.0 and 1:50 primary antibody overnight at 4°C (datasheet M02522). If staining differs, interpret the result alongside morphology and controls before attributing it to fixation, since the available tissue image does not establish a DGAT1-specific fixation effect (datasheet M02522).
Which intracellular DGAT1 pattern is plausible in chromogenic IHC?
Look first for cytoplasmic, finely reticular staining compatible with an endoplasmic reticulum membrane protein, rather than assuming a continuous cell-surface outline (UniProt O75907 subcellular location and topology). Tissue IHC describes general cytoplasmic expression, most abundant in small intestine and duodenum, with high staining in duodenal and small-intestinal endocrine cells (HPA tissue IHC). Use those cell populations as context, but score the compartment and cell identity separately because chromogenic signal alone cannot resolve an endoplasmic reticulum membrane (HPA tissue IHC; standard IHC practice). If nuclear-associated staining appears, note that HPA reports additional nucleolar and nucleolar-rim localization in ICC/IF, and require independent validation before treating that tissue pattern as specific (HPA subcellular).
Can DGAT1 topology or isoforms explain variable antibody access?
DGAT1 has 9 annotated transmembrane segments and alternating cytoplasmic and lumenal loops, so antibody access depends on the actual epitope location (UniProt O75907 topology). The supplied record lists no isoforms, but it does not identify the M02522 immunogen or epitope; therefore, do not infer which membrane face this antibody recognizes (UniProt O75907 isoforms; datasheet M02522). Compare retrieval in matched paraffin sections before changing primary concentration, starting with EDTA at pH 8.0 and the reported 1:50 incubation overnight at 4°C (datasheet M02522). For discordant staining, seek epitope documentation or independent antibody and genetic controls; phosphorylation at serines 17 and 18 alone does not establish epitope sensitivity (UniProt O75907 modified residues).
How can companion IF help assess DGAT1 cell identity and localization?
For a companion IF/ICC experiment, multiplex DGAT1 with an endocrine-cell identity marker to assess overlap within individual duodenal cells (HPA tissue IHC). Select spectrally separated fluorophores and inspect unstained tissue in every channel, because tissue autofluorescence can mimic a dim DGAT1 signal (standard IF practice). DGAT1 is an endoplasmic reticulum protein with cytoplasmic and lumenal loops, but the M02522 epitope is not specified; titrate permeabilisation according to verified epitope sidedness and preservation of membrane structure (UniProt O75907 topology; datasheet M02522). Evaluate an endoplasmic reticulum pattern alongside any nucleolar signal, which HPA also reports in ICC/IF (HPA subcellular).
What should I check when DGAT1 DAB staining looks diffuse?
First compare signal in expected cells against a no-primary section, then inspect whether background follows tissue edges or damaged areas (standard IHC practice). The M02522 image used 10% goat serum blocking, 1:50 primary overnight at 4°C, and an HRP/DAB readout (datasheet M02522). If diffuse brown staining dominates, optimize blocking, antibody concentration and wash stringency one variable at a time; include a peroxidase block as a general chromogenic IHC step (standard IHC practice). Assess whether the remaining pattern is cytoplasmic and cell-associated, as expected from ER localization and tissue IHC, rather than interpreting every brown deposit as DGAT1 (UniProt O75907 subcellular location; HPA tissue IHC).
How should I quantify DGAT1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical compartment and eligible cells before scoring, because DGAT1 staining varies by cell population in the supplied tissue profile (HPA tissue IHC). On matched paraffin sections, report the percentage of positive cells and an intensity-weighted H-score from 0 to 300, or positive-cell density per mm² (standard IHC scoring practice). Normalize to the number of evaluable cells or viable tissue area, and hold retrieval, 1:50 primary incubation and DAB development constant (datasheet M02522; standard IHC practice). Record background-subtracted thresholds and separate epithelial or endocrine-cell measurements from stromal signal so changes in tissue composition do not masquerade as altered DGAT1 expression (HPA tissue IHC; standard IHC scoring practice).
How can I distinguish DGAT1-positive cells from staining artefacts?
Treat cell-associated cytoplasmic staining in the appropriate anatomical compartment as more credible than diffuse edge, necrotic or lumenal deposits, given DGAT1's endoplasmic reticulum location and tissue IHC profile (UniProt O75907 subcellular location; HPA tissue IHC). High signal in duodenal endocrine cells is consistent with HPA, whereas adipocyte staining should prompt closer scrutiny because HPA reports adipocytes as not detected (HPA tissue IHC). Check no-primary and peroxidase-block controls before assigning brown DAB deposits to DGAT1, since endogenous peroxidase can produce chromogenic signal (standard IHC practice). HPA also reports nucleolar localization in ICC/IF, so investigate any isolated nuclear pattern with independent validation instead of automatically accepting or rejecting it (HPA subcellular).
Boster reagents
Best DGAT1 / Diacylglycerol O-acyltransferase 1 IHC Antibodies
The human-reactive IHC/IF catalog antibody M02522 has an IHC image from a paraffin-embedded human stomach cancer section (catalog: M02522 applications, reactivity, IHC caption).
Real IHC data
Anti-DGAT1 Rabbit Monoclonal Antibody
Cat # M02522
M02522 will render with its IHC image of a paraffin-embedded human stomach cancer section (catalog: M02522 IHC caption). IF/ICC is listed as an application for M02522, but the payload provides no IF image (catalog: M02522 applications, IF image alts).
Which to pick: Choose M02522 for human paraffin-section IHC: it is a rabbit monoclonal, and its own caption documents heat retrieval in EDTA at pH 8.0, primary antibody at 1:50, and DAB detection (catalog: M02522 host, clone, IHC caption). For IF/ICC, M02522 is application-listed at 1:50, without an IF figure in the payload (catalog: M02522 applications, IF dilution, IF image alts). Cross-species reactivity is unreported, and the paraffin-section IHC caption does not report the fixative (catalog: M02522 reactivity, IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75907 (DGAT1_HUMAN, Diacylglycerol O-acyltransferase 1).
- Human Protein Atlas. DGAT1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. DGAT1 subcellular location (ICC-IF): Mainly localized to the endoplasmic reticulum. In addition localized to the nucleoli and nucleoli rim..
- Human Protein Atlas. DGAT1 antibody validation summary (1 antibodies).
- Intestinal Failure and Aberrant Lipid Metabolism in Patients With DGAT1 Deficiency. Gastroenterology 2018 — PMC6058035.
- Lazy neutrophils - a lack of DGAT1 reduces the chemotactic activity of mouse neutrophils. Inflammation research : official journal of the European Histamine Research Society ... [et al.] 2024 — PMC11445369.
- Inhibition of diacylglycerol O-acyltransferase 1 provides neuroprotection by inhibiting ferroptosis in ischemic stroke. Molecular medicine (Cambridge, Mass.) 2025 — PMC12082899.
- Targeting DGAT1 Ameliorates Glioblastoma by Increasing Fat Catabolism and Oxidative Stress. Cell metabolism 2020 — PMC7415721.
- PubMed PMID:9756920 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:16214399 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DGAT1 (Diacylglycerol O-acyltransferase 1). Expected localisation General cytoplasmic staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M02522). Validated IHC antibody: M02522 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular 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 M02522); 2. Fixation: Image fixative and duration unreported (datasheet M02522); 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 M02522); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M02522); 8. Primary antibody: Rabbit monoclonal (clone 23D11) anti-DGAT1, 1:50 (datasheet M02522); 9. Primary incubation: Overnight at 4 °C (datasheet M02522). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M02522); Caveat: Adipocytes were not detected despite intestinal staining (HPA tissue IHC); Isoform / epitope: No annotated isoforms; 9 spans make epitope side relevant (UniProt); Regulation: No staining-linked regulator reported (UniProt). Evidence: UniProt O75907, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the endoplasmic reticulum. In addition localized…, Datasheet M02522, PMC6058035, PMC12082899. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DGAT1/immunohistochemistry-ihc-guide.