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RPN1 / Dolichyl-diphosphooligosaccharide--protein glycosyltransferase subunit 1 · IHC design guide
Design Immunohistochemistry for RPN1
Plan RPN1 chromogenic IHC in paraffin sections using its cytoplasmic tissue staining profile (HPA tissue IHC) and ER membrane topology (UniProt). Compare strongly stained Paneth or pancreatic exocrine cells with weaker muscle cells (HPA tissue IHC), and start antibody A05063-2 at 2–5 μg/ml (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RPN1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining (HPA tissue IHC); ER membrane (UniProt) | |
| Staining pattern | Cytoplasmic staining across tissues; intensity varies by cell type (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05063-2) | |
| Positive control | Duodenum+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Muscle cells may stain weakly despite broad tissue expression (HPA tissue IHC) | |
| Regulation | Broad tissue expression; no specific regulator reported (UniProt) | |
| Isoform / epitope | No isoforms annotated; lumenal 24–438, cytoplasmic 458–607 (UniProt) |
Section 1
Recommended RPN1 IHC & IF Protocols
The catalog antibody protocol is paired with two published RPN1 IHC protocols using paraffin sections (PMC11693735; PMC11201601).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A05063-2) |
| Fixation | Image fixative and duration unreported (datasheet A05063-2); 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 A05063-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05063-2) |
| Primary antibody | Rabbit anti-RPN1, 2-5 μg/ml (datasheet A05063-2) |
| Primary incubation | Overnight at 4 °C (datasheet A05063-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A05063-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RPN1-positive staining in paneth cells of duodenum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in all tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet A05063-2); citrate retrieval is a published alternative (PMC11201601).
Section 2
What Is the Expected RPN1 Staining Pattern?
RPN1 is an endoplasmic reticulum membrane protein with a lumenal region and cytoplasmic tail (UniProt P04843 topology). In paraffin sections, expect cytoplasmic staining across tissues, with stronger staining in selected cell types including pancreatic exocrine glandular cells and intestinal Paneth cells (HPA tissue IHC). HPA rates the tissue pattern Enhanced, while reporting medium consistency between antibody staining and RNA expression (HPA tissue IHC).
| Cytoplasmic staining is strongest in pancreatic exocrine glandular cells or intestinal Paneth cells (HPA tissue IHC). | This matches HPA's High staining in those cells and its broader cytoplasmic tissue pattern (HPA tissue IHC). Judge the distribution across cells as well as intensity; a strong color reaction alone does not establish specificity (general IHC practice). |
| Staining is predominantly nuclear, with little cytoplasmic signal. | A nucleus-dominant result conflicts with the reported cytoplasmic IHC pattern (HPA tissue IHC) and ER membrane location (UniProt P04843). Treat it as suspect and compare with a known-positive section and a primary-antibody omission control (general IHC practice). |
| Strong staining appears only in a cell population expected to stain weakly. | For example, HPA reports Low staining in cardiomyocytes and myocytes (HPA tissue IHC). Check cell identity and controls before calling the signal RPN1; cross-reactivity or endogenous detection activity can mimic target staining (general IHC practice). |
| Color spreads across tissue and empty areas without a clear cellular pattern. | That distribution does not resemble HPA's cytoplasmic cellular staining (HPA tissue IHC). Background may arise from nonspecific antibody binding or the detection system; inspect a primary-antibody omission control and review blocking and washing (general IHC practice). |
| A known-positive section shows no discernible cytoplasmic staining. | Pancreatic exocrine glandular cells and intestinal Paneth cells are reported High (HPA tissue IHC). If both sample and positive control fail, review the staining workflow before interpreting the sample as RPN1-negative (general IHC practice). |
💡Expected RPN1 appearanceCall positive when cytoplasmic staining is clear and relatively strong in a reported High cell population, such as pancreatic exocrine glandular cells (HPA tissue IHC); nucleus-dominant or control-matched color is suspect (UniProt P04843; general IHC practice).
| Tissue and cell selection | RPN1 is expressed in all tissues tested (UniProt P04843), while HPA reports Low tissue RNA specificity and cytoplasmic IHC staining across tissues (HPA tissue IHC). Use a reported High cell population as a positive reference; weak staining elsewhere does not by itself establish absence. |
| ER membrane topology | RPN1 has a lumenal segment at residues 24–438, a transmembrane segment at 439–457, and a cytoplasmic region at 458–607 (UniProt P04843 topology). The payload does not identify the catalog antibody's epitope, so topology cannot predict its retrieval response. |
| Processing and modification | UniProt annotates a signal peptide at residues 1–23, a chain at 24–607, and one glycosylation site at 299 (UniProt P04843). These annotations do not establish how fixation or antigen retrieval affects staining. |
| Interpretation of melanosome evidence | UniProt also lists melanosomes based on mass spectrometry of stage I–IV fractions (UniProt P04843). That finding does not replace the mainly ER localization observed by ICC-IF or justify calling a distinct melanosomal IHC pattern here (HPA subcellular). |
| Strength of tissue evidence | HPA labels the tissue IHC pattern Enhanced but describes only medium consistency with RNA expression (HPA tissue IHC). Its High and Low calls are reference patterns for interpretation, not guaranteed intensities in every section or staining run. |
| IF/ICC Q: What pattern belongs on its separate guide page? | A: Mainly ER localization, with additional cytosolic signal, is reported by HPA ICC-IF (HPA subcellular). That cellular localization helps assess compartment agreement; the IF/ICC workflow and protocol choices belong on its own guide page. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control tissue is blank or much weaker than expected. | A failed staining run or insufficient detection can obscure a reported High population; HPA's High calls are reference observations, not run guarantees (HPA tissue IHC; general IHC practice). | Confirm the expected cell population is present, then review retrieval, antibody application, detection, and counterstain against the laboratory's validated IHC workflow (general IHC practice). Do not infer a target-specific fixation effect. |
| Only nuclei stain strongly. | The distribution disagrees with cytoplasmic tissue IHC and ER localization (HPA tissue IHC; UniProt P04843). Nonspecific staining or detection background is possible (general IHC practice). | Compare a reported High cell population with a primary-antibody omission control; review blocking and detection if nuclear color persists without primary antibody (HPA tissue IHC; general IHC practice). |
| Muscle cells appear stronger than the chosen High control. | HPA reports Low staining in cardiomyocytes and skeletal myocytes, versus High staining in several glandular or Paneth cell populations (HPA tissue IHC). Cell assignment or assay background may explain the reversal (general IHC practice). | Verify morphology and compare both populations in the same run; use a primary-antibody omission control to assess detection-derived color (general IHC practice). Avoid declaring the muscle signal specific from intensity alone. |
| Color is diffuse across tissue or appears outside cells. | Diffuse color lacks the reported cytoplasmic cellular pattern (HPA tissue IHC). Nonspecific binding, inadequate washing, or endogenous detection activity can contribute (general IHC practice). | Inspect the omission control, wash and block according to the validated workflow, and address endogenous activity when relevant to the chromogenic detection chemistry (general IHC practice). Reassess only cell-associated signal. |
| Staining is present, but the cell compartment is hard to judge. | Dense chromogen or counterstain can obscure cytoplasmic boundaries (general IHC practice); HPA describes the tissue signal as cytoplasmic (HPA tissue IHC). | Review a less saturated field and the counterstain, and compare with a reported High cell population (general IHC practice; HPA tissue IHC). Score localization only where cells remain distinguishable. |
| The slide suggests a distinct melanosomal pattern. | UniProt's melanosome annotation comes from fraction mass spectrometry, whereas HPA reports mainly ER localization in ICC-IF (UniProt P04843; HPA subcellular). | Prioritize the observed cytoplasmic IHC pattern and check controls before assigning a finer compartment (HPA tissue IHC; general IHC practice). Treat melanosomal identity as unresolved by this IHC evidence. |
Experiment evidence
IHC & IF images showing RPN1 staining
A05063-2 · IHCIHC analysis of RPN1 using anti-RPN1 antibody (A05063-2). RPN1 was detected in a paraffin-embedded section of human liver cancer tissue. Hea
A05063-2 · IHCIHC analysis of RPN1 using anti-RPN1 antibody (A05063-2). RPN1 was detected in a paraffin-embedded section of human lung adenocarcinoma tiss
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000163902-RPN1/tissue/cerebral+cortex#imid_18230079
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000163902-RPN1/tissue/colon#imid_18229989
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000163902-RPN1/tissue/liver#imid_18230006
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000163902-RPN1/tissue/kidney#imid_18229991
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA026828; green = RPN1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA026828; green = RPN1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RPN1 IHC & IF
🧪Run pancreas first: exocrine glandular cells should stain strongly (HPA: High in pancreatic exocrine glandular cells). HPA detects RPN1 in all 45 scored tissues, so there is no validated negative tissue; no cell population on the positive slide is established as RPN1-negative, and the no-primary and isotype controls should define background staining (HPA: no negative tissue rows).
Positive control tissue: Duodenum (Paneth cells, HPA High)
Negative control tissue: None in HPA: RPN1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RPN1 in A-431, U2OS, U-251MG, NIH 3T3, with annotated localisation: Endoplasmic reticulum (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a rabbit IgG isotype control matched to the primary antibody’s clonality where known (selected-SKU caption: rabbit anti-RPN1 primary). Use RPN1 knockout material processed in parallel as a biological negative; for pancreatic HRP–DAB staining, quench endogenous peroxidase and assess any residual background (selected-SKU caption: HRP–DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state a fixative (selected-SKU caption: fixative not stated). Start with heat-mediated EDTA retrieval at pH 8.0 as reported for that IHC example, while treating retrieval dependence as unestablished (selected-SKU caption: EDTA pH 8.0 retrieval). The supplied evidence does not show that frozen sections or IF are easier than paraffin IHC; in pancreas, assess acinar preservation because autolysis can obscure staining patterns (HPA: High in pancreatic exocrine glandular cells; standard histology practice).
HPA tissue IHC evidence for RPN1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Duodenum | Paneth cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Pancreas | Exocrine glandular cells | High | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | High | Protein (IHC) | HPA → |
| Skin | Fibrohistiocytic cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: RPN1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced RPN1 IHC Tips
Use the catalog antibody’s paraffin-section IHC conditions as the starting point, then judge staining against RPN1’s expected cellular distribution and tissue controls.
Which retrieval conditions should I start with for RPN1 in paraffin sections?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin-section RPN1 IHC (datasheet A05063-2). The catalog image used these conditions on a human liver cancer section, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet A05063-2). If staining is weak, compare retrieval duration on adjacent sections while holding antibody concentration and detection constant; excessive heating can damage morphology (standard IHC practice). Evaluate improvement as intracellular cytoplasmic staining with preserved tissue structure, since tissue IHC shows cytoplasmic expression and RPN1 is mainly associated with the endoplasmic reticulum (HPA tissue IHC; HPA subcellular).
How should I troubleshoot fixation-related loss of RPN1 staining?
Target-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not state a fixative, and its retrieval result cannot establish which fixation conditions preserve RPN1 (datasheet A05063-2). Record the fixative and fixation duration for each specimen, then compare matched sections processed with the same EDTA pH 8.0 retrieval and detection conditions (standard IHC practice; datasheet A05063-2). Check morphology alongside staining, because poor preservation or uneven processing can make regional signal difficult to interpret (standard IHC practice). Use an internal tissue control when available; HPA reports high staining in pancreatic exocrine glandular cells and low staining in cardiomyocytes (HPA tissue IHC).
Where should convincing RPN1 staining appear in tissue sections?
Expect a predominantly intracellular, cytoplasmic pattern in chromogenic sections, consistent with RPN1’s endoplasmic reticulum membrane location and HPA tissue IHC profile (UniProt P04843 subcellular; HPA tissue IHC). A fine perinuclear or reticular distribution may support that assignment when section quality permits, while HPA subcellular imaging also reports an additional cytosolic location (HPA subcellular). Assess staining within intact cells and compare nearby positive and low-expression cell populations before calling a focal deposit specific (standard IHC practice; HPA tissue IHC). UniProt also reports RPN1 in melanosome fractions by mass spectrometry, but that finding alone does not establish a distinct chromogenic staining pattern in tissue (UniProt P04843 subcellular).
Could topology or processing explain discordant RPN1 staining?
RPN1 has no annotated isoforms in this record, so an isoform-specific explanation for differing section patterns is unsupported (UniProt P04843 isoforms). Its signal peptide spans residues 1–23, the mature chain begins at 24, and its membrane segment spans 439–457 (UniProt P04843 processing and topology). The 24–438 region is lumenal and 458–607 is cytoplasmic; the recorded glycosylation site is residue 299 (UniProt P04843 topology and glycosylation). Because the catalog antibody’s epitope is unspecified here, request its immunogen or epitope details before attributing staining changes to processing, glycosylation, or accessibility (datasheet A05063-2; standard IHC practice).
How can IF help resolve ambiguous RPN1 staining seen by IHC?
Use IF as a follow-up to the chromogenic section result, checking whether RPN1 signal overlaps an independently validated marker of the cell type being assessed; HPA reports high tissue staining in Paneth cells and pancreatic exocrine glandular cells (HPA tissue IHC). Choose a fluorophore channel with low background in that specimen and inspect single-channel images, because tissue autofluorescence can imitate specific IF signal (standard IF practice). Plan permeabilisation around the mapped antibody epitope: RPN1 residues 24–438 are lumenal and 458–607 are cytoplasmic (UniProt P04843 topology). Confirm epitope information before choosing conditions, since it is not supplied for the catalog antibody here (datasheet A05063-2).
How do I reduce diffuse or patchy DAB background?
The selected paraffin-section image used a 10% goat-serum block, a peroxidase-conjugated secondary, and DAB development (datasheet A05063-2). Include a primary-omission control and a general endogenous-peroxidase blocking step to distinguish detection background from antibody-associated staining (standard IHC practice). Compare nearby sections while adjusting blocking, wash stringency, or DAB development one variable at a time; retain the documented 2 μg/ml primary concentration as the initial reference (standard IHC practice; datasheet A05063-2). Score only signal within intact cellular profiles, since RPN1 tissue staining is reported as cytoplasmic and its main subcellular location is the endoplasmic reticulum (HPA tissue IHC; HPA subcellular).
How should I score heterogeneous RPN1 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then report the percentage of positive cells alongside staining intensity or an H-score calculated from intensity categories 0–3 (standard IHC practice). Keep segmentation and positivity thresholds fixed across sections, and normalise positive-cell counts to the number of eligible cells or a measured area in mm² (standard IHC practice). Exclude necrotic tissue, folds, and section edges from the denominator using the same rules for every specimen (standard IHC practice). Interpret between-tissue differences cautiously: HPA describes cytoplasmic expression across tissues, low RNA tissue specificity, and only medium consistency between staining and RNA expression (HPA tissue IHC).
What distinguishes true RPN1 signal from an artefact?
A credible positive shows intracellular cytoplasmic staining in preserved cells, matching the HPA tissue profile and RPN1’s main endoplasmic reticulum location (HPA tissue IHC; HPA subcellular). Check whether staining follows expected cell populations: HPA reports high signal in placental trophoblastic cells and low signal in adipocytes, although those comparisons do not validate an individual section (HPA tissue IHC). Treat staining confined to cut edges, folds, or necrotic zones as suspect, and compare it with a primary-omission control for detection background (standard IHC practice). Diffuse nuclear staining or a deposit outside cells requires further validation against morphology and controls before being assigned to RPN1 (UniProt P04843 subcellular; standard IHC practice).
Boster reagents
Best RPN1 / Dolichyl-diphosphooligosaccharide--protein glycosyltransferase subunit 1 IHC Antibodies
A05063-2 has human paraffin-section IHC images (A05063-2 IHC captions). M05063-1 lists human and rat reactivity and IF/ICC use, with no supplied image captions (M05063-1 catalog).
Real IHC data
Anti-RPN1 Antibody ®
Cat # A05063-2
A05063-2 has IHC images from human liver cancer, lung adenocarcinoma, placenta and rectum adenocarcinoma paraffin sections (A05063-2 IHC captions). M05063-1 lists human and rat reactivity and IHC and IF/ICC applications; no IHC or IF image captions are supplied (M05063-1 catalog).
Which to pick: For human tissue IHC, choose A05063-2 for its paraffin-section images; its captions report EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody, but do not report the fixative (A05063-2 IHC captions). For IF/ICC, choose M05063-1 because those applications are listed for the rabbit monoclonal clone 32R32, although no IF image is supplied (M05063-1 catalog). M05063-1 is also the listed option for rat samples; A05063-2 lists human reactivity only (M05063-1 catalog; A05063-2 catalog).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P04843 (RPN1_HUMAN, Dolichyl-diphosphooligosaccharide--protein glycosyltransferase subunit 1).
- Human Protein Atlas. RPN1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. RPN1 subcellular location (ICC-IF): Mainly localized to the endoplasmic reticulum. In addition localized to the cytosol..
- Human Protein Atlas. RPN1 antibody validation summary (3 antibodies).
- Dissecting the functions and regulatory mechanisms of disulfidoptosis-related RPN1 in pan-cancer: modulation of immune microenvironment and cellular senescence. Frontiers in immunology 2024 — PMC11693735.
- Multi-Omics Insights into Disulfidptosis-Related Genes Reveal RPN1 as a Therapeutic Target for Liver Cancer. Biomolecules 2024 — PMC11201601.
- Integrated analysis of disulfidptosis-related genes SLC7A11, SLC3A2, RPN1 and NCKAP1 across cancers. Discover oncology 2024 — PMC11607372.
- Knockdown of Oligosaccharyltransferase Subunit Ribophorin 1 Induces Endoplasmic-Reticulum-Stress-Dependent Cell Apoptosis in Breast Cancer. Frontiers in oncology 2021 — PMC8578895.
- PubMed PMID:3034581 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RPN1 (Dolichyl-diphosphooligosaccharide--protein glycosyltransferase subunit 1). Expected localisation Cytoplasmic staining (HPA tissue IHC); ER membrane (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05063-2). Validated IHC antibody: A05063-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Duodenum (Paneth cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A05063-2); 2. Fixation: Image fixative and duration unreported (datasheet A05063-2); 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 A05063-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05063-2); 8. Primary antibody: Rabbit anti-RPN1, 2-5 μg/ml (datasheet A05063-2); 9. Primary incubation: Overnight at 4 °C (datasheet A05063-2). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Muscle cells may stain weakly despite broad tissue expression (HPA tissue IHC); Isoform / epitope: No isoforms annotated; lumenal 24–438, cytoplasmic 458–607 (UniProt); Regulation: Broad tissue expression; no specific regulator reported (UniProt). Evidence: UniProt P04843, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the endoplasmic reticulum. In addition localized…, Datasheet A05063-2, PMC11693735, PMC11201601. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RPN1/immunohistochemistry-ihc-guide.