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DPAGT1 / UDP-N-acetylglucosamine--dolichyl-phosphate N-acetylglucosaminephosphotransferase · IHC design guide
Design Immunohistochemistry for DPAGT1
Plan DPAGT1 IHC-P around granular cytoplasmic staining, with colon glandular cells as a high-staining control (HPA tissue IHC). This guide covers fixation, retrieval, chromogenic detection and scoring, starting the IHC-validated antibody at 2.5 μg/mL (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DPAGT1 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 | Granular cytoplasm (HPA tissue IHC); ER membrane (UniProt) | |
| Staining pattern | Often granular cytoplasm in glandular and epithelial cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+3 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin tissue 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 A08142) | |
| Caveat | Staining and RNA show only medium consistency (HPA tissue IHC) | |
| Regulation | No specific regulator annotated (UniProt) | |
| Isoform / epitope | 3 isoforms; lumenal vs cytoplasmic epitope matters (UniProt) |
Section 1
Recommended DPAGT1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by a published paraffin-section protocol for human tongue squamous cell carcinoma (PMC7758185).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A08142); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-DPAGT1, 2.5 μg/mL (datasheet A08142) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DPAGT1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression, often in a granular pattern. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen retrieval); the published protocol uses autoclaving instead (PMC7758185).
Section 2
What Is the Expected DPAGT1 Staining Pattern?
DPAGT1 is a 10-pass endoplasmic reticulum membrane protein (UniProt Q9H3H5 topology). In paraffin-section IHC, expect cytoplasmic staining, often granular, in cells such as colon and duodenal glandular cells, bronchial ciliated-cell bodies, and adrenal glandular cells (HPA tissue IHC: High; general cytoplasmic, often granular profile). HPA rates its tissue staining Approved, with medium consistency between antibody staining and RNA expression (HPA tissue IHC: reliability).
| Granular cytoplasmic signal in colon or duodenal glandular cells, with little signal in nearby unstained structures. | This matches HPA's general cytoplasmic, often granular profile and its High staining in these glandular cells (HPA tissue IHC). The pattern is compatible with DPAGT1's ER membrane location; chromogenic IHC alone does not resolve individual ER membranes (UniProt Q9H3H5; general IHC practice). |
| Predominantly nuclear, cell-surface, or extracellular staining with little cytoplasmic signal. | Treat this as a compartment mismatch: UniProt places DPAGT1 in the ER membrane, while HPA reports a cytoplasmic tissue pattern (UniProt Q9H3H5; HPA tissue IHC). Review controls and staining distribution before assigning such signal to DPAGT1 (general IHC practice). |
| Strong staining in adipocytes, smooth muscle cells, ovarian follicle cells, or chondrocytes. | HPA reports DPAGT1 as Not detected in those cell types (HPA tissue IHC). Unexpected staining raises possible cross-reactivity or endogenous detection activity; a single discrepant section cannot establish either cause, and Not detected does not prove protein absence (general IHC interpretation). |
| Diffuse color across cells, stroma, and slide background without a recognizable granular cytoplasmic pattern. | This is less convincing than HPA's often granular cytoplasmic profile (HPA tissue IHC). Widespread nonspecific antibody binding, inadequate blocking, or endogenous detection activity are possible technical explanations to test with controls (general IHC practice). |
| No signal in a well-preserved colon or duodenal glandular-cell section. | HPA reports High staining in these cells, so absence of signal calls for a run-level check (HPA tissue IHC). Review the catalog antibody's IHC-P conditions, retrieval, detection reagents, and a positive control before interpreting the specimen as negative (general IHC practice). |
💡Expected DPAGT1 appearanceCall a result positive when glandular cells of colon or duodenum show convincing cytoplasmic, often granular chromogenic signal consistent with HPA's High staining and UniProt's ER membrane location; isolated nuclear, extracellular, or widespread diffuse color is suspect (HPA tissue IHC; UniProt Q9H3H5; general IHC interpretation).
| Compartment and topology | DPAGT1 has 10 transmembrane segments and an ER membrane location (UniProt Q9H3H5 topology). Interpret its predicted location against HPA's cytoplasmic, often granular IHC profile; routine chromogenic resolution does not prove ER colocalization (HPA tissue IHC; general IHC practice). |
| Tissue choice and expression range | HPA reports High staining in several epithelial or glandular populations, Low staining in selected others, and Not detected in specific cell types; its RNA summary is low tissue specificity (HPA tissue IHC). Compare named cell populations rather than labeling a whole organ uniformly positive or negative (general IHC interpretation). |
| Antibody evidence | HPA lists HPA053878 as IHC Approved and describes tissue-level agreement with RNA as medium consistency (HPA antibodies; HPA tissue IHC). This supports cautious pattern comparison; the supplied record does not report IHC Enhanced validation or an epitope, so neither independent-antibody concordance nor epitope accessibility can be assumed (HPA antibodies; supplied record). |
| Isoforms and processing | UniProt lists 3 isoforms, one chain spanning residues 1–408, and no annotated signal peptide or propeptide (UniProt Q9H3H5). Because the supplied antibody record gives no epitope, this section cannot determine which isoforms its IHC signal represents (HPA antibodies; supplied record). |
| Can IF/ICC confirm this pattern? | UniProt's ER membrane assignment makes intracellular, ER-like fluorescence a plausible comparison, but HPA's ICC-IF entry says only Membrane, gives no main location, and lists no cell-line images (UniProt Q9H3H5; HPA subcellular). It therefore supplies no observed ER colocalization or IF/ICC validation here (HPA subcellular; HPA antibodies). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected cytoplasmic signal is absent in a High-staining control tissue (HPA tissue IHC). | An IHC run, retrieval, antibody, or detection failure is possible; the slide alone cannot identify which (general IHC practice). | Check the catalog antibody's IHC-P instructions and run a suitable positive control through the same retrieval and detection steps (general IHC practice). |
| Only nuclei or extracellular material stain. | The dominant compartment conflicts with UniProt's ER membrane location and HPA's cytoplasmic IHC profile (UniProt Q9H3H5; HPA tissue IHC). | Inspect staining morphology and controls, then reassess antibody specificity and detection background before scoring the result (general IHC practice). |
| Not-detected reference cell types show strong color (HPA tissue IHC). | Possible antibody cross-reactivity or endogenous detection activity; HPA's Not detected designation is an observed staining result, not proof of molecular absence (HPA tissue IHC; general IHC interpretation). | Compare a no-primary control and the expected positive-cell pattern; investigate background before calling those cells DPAGT1-positive (general IHC practice). |
| Color covers stroma and many unrelated cell populations. | Nonspecific binding or endogenous chromogenic activity may obscure HPA's often granular cytoplasmic profile (HPA tissue IHC; general IHC practice). | Review blocking and wash steps, use a no-primary control, and evaluate signal against cell boundaries and morphology (general IHC practice). |
| A weak tissue is negative while a High-staining control is positive (HPA tissue IHC). | HPA reports Low staining in some cell types, so a negative call in a weak sample may reflect limited assay sensitivity (HPA tissue IHC; general IHC interpretation). | Score the control and sample separately; record the weak sample as not detected under the tested conditions rather than inferring DPAGT1 is absent (general IHC interpretation). |
| An IF/ICC image seems to show a precise ER pattern. | The supplied HPA ICC-IF entry has no cell-line images or specified main location; ER localization comes from UniProt (HPA subcellular; UniProt Q9H3H5). | Describe the observed fluorescence separately from the UniProt prediction, and require independent colocalization evidence before claiming ER-specific IF staining (general IF practice). |
Experiment evidence
IHC & IF images showing DPAGT1 staining
A08142 · IHCImmunohistochemistry of DPAGT1 in human kidney tissue with DPAGT1 antibody at 2.5 μg/mL.
A08142 · IFImmunofluorescence of DPAGT1 in human kidney tissue with DPAGT1 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000172269-DPAGT1/tissue/cerebral+cortex#imid_15141558
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000172269-DPAGT1/tissue/colon#imid_15141476
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000172269-DPAGT1/tissue/liver#imid_15141469
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000172269-DPAGT1/tissue/kidney#imid_15141486
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DPAGT1 IHC & IF
🧪Run colon first and look for staining in glandular cells (HPA: High in colon glandular cells); run adipose tissue as the negative comparator and expect adipocytes to remain unstained (HPA: Not detected in adipocytes). On the colon slide, use morphologically distinct cells without staining to assess local background, without treating them as a validated DPAGT1-negative population (standard IHC practice).
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 carries no ICC-IF cell line for DPAGT1; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; an isotype control matched to the primary antibody’s host species, clonality and concentration; and, if available, DPAGT1-knockout tissue or an immunizing-peptide block as a biological specificity check (standard IHC practice). For chromogenic colon IHC, block endogenous peroxidase and inspect the no-primary slide for background, especially in inflammatory cells (standard IHC practice).
⚠️Feasibility: No DPAGT1-specific fixation window or fixation effect is reported in the supplied evidence, and the selected A08142 kidney IHC caption does not state its fixative (A08142 tissue-IHC caption). Antigen-retrieval dependency is unreported; compare retrieval conditions empirically on paraffin sections and assess whether staining is compatible with ER localization (UniProt Q9H3H5: endoplasmic reticulum membrane; standard IHC practice). The supplied evidence does not establish whether frozen sections or IF/ICC are easier; endogenous peroxidase in colon inflammatory cells can complicate chromogenic interpretation (standard IHC practice).
HPA tissue IHC evidence for DPAGT1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Ciliated cells (cell body) | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | Follicle cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Chondrocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced DPAGT1 IHC Tips
Troubleshoot DPAGT1 staining in paraffin section IHC using the specified retrieval method, membrane topology, and reported tissue staining patterns (UniProt Q9H3H5; HPA tissue IHC).
How should I retrieve DPAGT1 when staining is weak?
Use citrate pH 6.0 heat induced epitope retrieval at 95–98 °C for 20 min (page retrieval rule: cytoplasmic / membrane antigen). Allow sections to cool in retrieval buffer, then rinse consistently before blocking so differences in heating do not masquerade as biological variation (standard IHC practice). DPAGT1 crosses the endoplasmic reticulum membrane 10 times, so epitope accessibility may depend on section processing and the antibody’s epitope (UniProt Q9H3H5 topology; standard IHC practice). If staining remains weak, compare a controlled increase in retrieval time on adjacent sections while checking tissue integrity and background; the supplied evidence does not establish an antibody specific alternative buffer (standard IHC practice; supplied retrieval rule).
Could fixation explain variable DPAGT1 staining between blocks?
Target specific fixation sensitivity is unknown from the supplied evidence; the selected kidney image caption gives antibody concentration but does not state its fixative (A08142 tissue IHC caption). Record fixative, fixation duration, processing history, and section age for each block before attributing differences to DPAGT1 biology (standard IHC practice). Compare adjacent sections with the same citrate pH 6.0, 95–98 °C, 20 min retrieval and identical detection conditions to isolate block effects (page retrieval rule; standard IHC practice). Use an internal tissue control and inspect morphology, since damaged or unevenly fixed areas can distort chromogenic intensity and the fraction of cells scored positive (standard IHC practice).
Where should credible DPAGT1 chromogenic staining appear?
Expect predominantly cytoplasmic, often granular staining in tissue IHC, consistent with DPAGT1 residing in the endoplasmic reticulum membrane (HPA tissue IHC profile; UniProt Q9H3H5 subcellular location). Evaluate staining within the cell body rather than treating a sharply nuclear or extracellular signal as expected localisation (HPA tissue IHC profile; standard IHC interpretation). DPAGT1 has 10 annotated transmembrane segments, so a chromogenic deposit need not resolve an individual membrane or distinguish its cytoplasmic and lumenal surfaces (UniProt Q9H3H5 topology; standard IHC practice). Compare the pattern across intact cells and adjacent sections, and use morphology to separate granular cytoplasm from precipitate or pigment (standard IHC practice).
Can the IHC antibody distinguish DPAGT1 isoforms or epitope orientation?
DPAGT1 has 3 annotated isoforms, but the supplied antibody evidence does not identify an epitope or establish isoform selectivity (UniProt Q9H3H5 isoforms; supplied A08142 caption). Its 10 transmembrane segments place short regions on alternating cytoplasmic and lumenal sides, while residue 146 is an annotated glycosylation site (UniProt Q9H3H5 topology and glycosylation). Consult the antibody’s documented immunogen or epitope before predicting which isoforms or membrane faces it can detect (standard IHC practice). Until that information and appropriate validation are available, report staining as DPAGT1 immunoreactivity rather than assigning it to a specific isoform, accessible loop, or glycosylation state (standard IHC interpretation).
How can IF help assess the DPAGT1 IHC pattern?
Use IF as a complementary localisation check; the supplied subcellular record lists no cell line ICC/IF images for DPAGT1 (HPA subcellular record). Multiplex DPAGT1 with a marker for the expected cell population, such as glandular cells in colon, and verify each channel separately before interpreting overlap (HPA tissue IHC: High in colon glandular cells; standard IF practice). Choose fluorophores in channels with low tissue autofluorescence and include unstained and single label controls (standard IF practice). Because DPAGT1 spans the endoplasmic reticulum membrane 10 times, set permeabilisation according to the documented epitope’s cytoplasmic or lumenal orientation; that orientation is not supplied for this antibody (UniProt Q9H3H5 topology; supplied A08142 caption; standard IF practice).
What should I check when DPAGT1 IHC shows diffuse brown background?
First inspect no primary and detection only controls to identify nonspecific chromogen deposition or endogenous peroxidase activity (standard chromogenic IHC practice). Apply a peroxidase block when using HRP and DAB, and optimise protein blocking and washes according to the detection system (standard chromogenic IHC practice). Titrate the IHC validated antibody around the selected kidney image’s reported 2.5 µg/mL concentration while assessing specific cytoplasmic contrast; that caption does not establish an optimum for every tissue (A08142 tissue IHC caption; HPA tissue IHC profile). If stronger retrieval increases both granular cell body signal and diffuse haze, compare adjacent sections under matched detection and inspect tissue edges for artefact (page retrieval rule; standard IHC practice).
How should DPAGT1 staining be scored across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and scoring region before measuring DPAGT1, since HPA reports high staining in colon glandular cells but no detection in adipocytes (HPA tissue IHC). For chromogenic IHC, record the percentage of positive target cells and an intensity based H score, or positive cell density per mm² when cell counts are the primary outcome (standard IHC quantification practice). Normalise to the number or area of viable, morphologically identifiable target cells, and exclude folds, edges, and necrotic regions (standard IHC quantification practice). Keep retrieval, exposure to chromogen, thresholds, and counterstain consistent across sections, then report these settings with representative images (standard IHC practice).
How do I separate true DPAGT1 signal from artefact?
A plausible positive pattern is cytoplasmic and often granular, matching the reported tissue profile and endoplasmic reticulum membrane location (HPA tissue IHC profile; UniProt Q9H3H5 subcellular location). Judge it within identified cells: HPA reports high signal in adrenal glandular cells and no detection in adipocytes, although its tissue IHC reliability is Approved with medium consistency against RNA (HPA tissue IHC). Treat exclusively nuclear or extracellular staining, tissue edge accentuation, and signal confined to necrosis as reasons to recheck specificity and morphology (HPA tissue IHC profile; standard IHC interpretation). Compare no primary controls for endogenous enzyme signal and matched adjacent sections for reproducibility before assigning a biological difference (standard chromogenic IHC practice).
Boster reagents
Best DPAGT1 / UDP-N-acetylglucosamine--dolichyl-phosphate N-acetylglucosaminephosphotransferase IHC Antibodies
The catalog lists one anti-DPAGT1 antibody with IHC and IF images from human kidney tissue (catalog: image captions); human and mouse reactivity is listed (catalog: reactivity).
Real IHC data
Anti-DPAGT1 Antibody
Cat # A08142
A08142 will render with a human kidney IHC image at 2.5 μg/mL (catalog: IHC image caption). A08142 also has a human kidney IF image at 20 μg/mL (catalog: IF image caption).
Which to pick: Choose A08142 for paraffin-section IHC (catalog: IHC-P application); its IHC caption shows human kidney tissue, but does not report the fixative (catalog: IHC image caption). Choose A08142 for IF; ICC validation is not listed (catalog: applications). For work involving both human and mouse samples, A08142 lists reactivity with both species, although the supplied IHC and IF images show human tissue only (catalog: reactivity and image captions); no clone is specified.
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9H3H5 (GPT_HUMAN, UDP-N-acetylglucosamine--dolichyl-phosphate N-acetylglucosaminephosphotransferase).
- Human Protein Atlas. DPAGT1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. DPAGT1 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. DPAGT1 antibody validation summary (1 antibodies).
- NFAT5 promotes oral squamous cell carcinoma progression in a hyperosmotic environment. Laboratory investigation; a journal of technical methods and pathology 2021 — PMC7758185.
- O-GlcNAcylation modulates expression and abundance of N-glycosylation machinery in an inherited glycosylation disorder. Cell reports 2024 — PMC11656453.
- Mechanisms of Sodium/Iodide Symporter-Mediated Mammary Gland Iodine Compensation during Lactation. Nutrients 2022 — PMC9460413.
- A Dpagt1 Missense Variant Causes Degenerative Retinopathy without Myasthenic Syndrome in Mice. International journal of molecular sciences 2022 — PMC9570038.
- PubMed PMID:9451016 — UniProt-cited evidence.
- PubMed PMID:29459785 — UniProt-cited evidence.
- PubMed PMID:30388443 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DPAGT1 (UDP-N-acetylglucosamine--dolichyl-phosphate N-acetylglucosaminephosphotransferase). Expected localisation Granular cytoplasm (HPA tissue IHC); ER membrane (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A08142 at 2.5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A08142); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-DPAGT1, 2.5 μg/mL (datasheet A08142); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin tissue 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 A08142); Caveat: Staining and RNA show only medium consistency (HPA tissue IHC); Isoform / epitope: 3 isoforms; lumenal vs cytoplasmic epitope matters (UniProt); Regulation: No specific regulator annotated (UniProt). Evidence: UniProt Q9H3H5, HPA tissue IHC · Approved, HPA ICC-IF · Membrane, Datasheet A08142, PMC7758185. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DPAGT1/immunohistochemistry-ihc-guide.