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ARL6IP1 / ADP-ribosylation factor-like protein 6-interacting protein 1 · IHC design guide
Design Immunohistochemistry for ARL6IP1
Plan chromogenic ARL6IP1 IHC-P with the catalog antibody starting at 2 μg/mL (datasheet: IHC-P). Expect cytoplasmic tissue staining (HPA tissue IHC), with ER membrane localisation as molecular context (UniProt); interpret intensity cautiously because tissue staining reliability is uncertain (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ARL6IP1 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 membrane (UniProt) | |
| Staining pattern | Duodenal glandular cells: cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Duodenum+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 fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining may detect proteins from more than one gene (HPA tissue IHC) | |
| Regulation | Highest in early myeloid progenitors (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unreported (UniProt) |
Section 1
Recommended ARL6IP1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by one published ARL6IP1 IHC protocol using human oral squamous cell carcinoma sections (PMC13383874).
| Sample | human bladder tissue; fixation not specified (datasheet A08487) |
| Fixation | Image fixative and duration unreported (datasheet A08487); 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-ARL6IP1, 2 μg/mL (datasheet A08487) |
| 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 | ARL6IP1-positive staining in glandular cells of duodenum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval); the published protocol also uses citrate pH 6.0 (PMC13383874).
Section 2
What Is the Expected ARL6IP1 Staining Pattern?
ARL6IP1 is a membrane protein concentrated in ER tubules and sheet edges; expect predominantly cytoplasmic staining rather than a nuclear pattern (UniProt Q15041 localization and topology). In paraffin sections, HPA reports high staining in duodenal and small-intestinal glandular cells, among others, but rates tissue IHC reliability Uncertain because staining and RNA data have medium consistency and the antibody may target proteins from more than one gene (HPA tissue IHC).
| Granular or reticular cytoplasmic stain in intestinal glandular cells, with little nuclear signal. | This fits the reported cytoplasmic tissue pattern and ER localization (HPA tissue IHC; UniProt Q15041 localization). High staining in duodenum and small intestine makes these useful comparison tissues, but a matching image alone does not establish antibody specificity because HPA rates the tissue evidence Uncertain (HPA tissue IHC). |
| Strong, sharply nuclear staining dominates the section. | Nuclear dominance is discordant with the reported ER membrane location and cytoplasmic tissue pattern (UniProt Q15041 localization; HPA tissue IHC). Treat it as a possible staining artifact or off-target signal; review morphology, detection controls and independent antibody evidence before assigning it to ARL6IP1. |
| The strongest signal appears in an unexpected cell population while the expected glandular cells remain weak. | For duodenum or small intestine, this reverses the HPA high-glandular-cell pattern (HPA tissue IHC). Possible explanations include cross-reactivity or endogenous detection activity. Because HPA cautions that the antibody may recognize products of more than one gene, confirm cell identity and compare suitable detection controls (HPA tissue IHC). |
| Color is widespread across cells, stroma and empty areas without a clear cellular pattern. | This does not resolve the reported cytoplasmic distribution (HPA tissue IHC). In routine chromogenic IHC, widespread haze can arise from nonspecific antibody binding, inadequate blocking, residual detection activity or excessive development. Inspect a primary-antibody-omission control and adjust one workflow variable at a time. |
| A duodenal or small-intestinal section shows no glandular-cell signal. | HPA records high staining in those glandular cells, so a blank section warrants a technical check (HPA tissue IHC). Verify tissue identity, section integrity, retrieval and detection performance using appropriate controls. Do not infer biological absence from one negative stain, especially given HPA's Uncertain tissue reliability (HPA tissue IHC). |
💡Expected ARL6IP1 appearanceCall a result provisionally positive when glandular cells in duodenum or small intestine show clear cytoplasmic, ER-consistent staining near HPA's reported high level; dominant nuclear or acellular color is suspect, and specificity remains uncertain (HPA tissue IHC; UniProt Q15041 localization).
| Membrane topology and access | ARL6IP1 has three transmembrane segments, with annotated cytoplasmic and lumenal regions (UniProt Q15041 topology). Epitope access could depend on antibody binding site and section processing; the supplied records do not identify the epitope or establish a target-specific antigen-retrieval condition. |
| Tissue choice and conflicting evidence | HPA reports high staining in cardiomyocytes, whereas UniProt says ARL6IP1 was not detected in heart; HPA also reports no staining in bone-marrow hematopoietic cells despite UniProt expression in hematopoietic lineages (HPA tissue IHC; UniProt Q15041 tissue specificity). Avoid treating either tissue as a definitive positive or negative control. |
| Antibody evidence | HPA lists tissue IHC as Uncertain and antibody HPA045307 as IHC Uncertain; its tissue summary cautions that antibodies may target proteins from more than one gene (HPA tissue IHC; HPA antibody validation). Interpret even correctly located signal provisionally and seek independent confirmation for consequential conclusions. |
| Isoforms and processing | UniProt lists three isoforms, one chain spanning residues 1–203, no annotated signal peptide or propeptide, and no glycosylation sites (UniProt Q15041 isoforms and processing). These annotations do not show which isoforms the IHC antibody recognizes or support an expected shedding pattern. |
| IF/ICC: what localization should appear? | An ER pattern is the supported subcellular result; HPA shows ICC-IF images from A-431, U-251MG and U2OS and rates HPA045307 ICC Supported (HPA subcellular; HPA antibody validation). HPA cautions that this localization evidence involves antibodies targeting proteins from multiple genes (HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| High signal in nuclei rather than cytoplasm. | The location conflicts with ER localization and reported cytoplasmic tissue staining (UniProt Q15041 localization; HPA tissue IHC). | Check the primary-antibody-omission control, counterstain and cell boundaries; repeat with an independent antibody if available. Record the pattern as unresolved until specificity is supported. |
| No glandular staining in duodenum or small intestine. | These cells have high HPA tissue staining, but that evidence is rated Uncertain (HPA tissue IHC). A technical failure is possible. | Confirm section identity and morphology, then check the run's positive and detection controls. Review retrieval and dilution as general IHC variables without assuming a known ARL6IP1-specific setting. |
| Diffuse chromogen obscures cell boundaries. | Nonspecific binding, residual endogenous detection activity or prolonged development can create broad background in chromogenic IHC. | Compare a primary-antibody-omission section; review blocking, washes and development time. Score localization only after cellular boundaries are interpretable. |
| Cardiomyocytes stain strongly but another source suggests no heart expression. | HPA reports high cardiomyocyte staining, while UniProt reports no detection in heart (HPA tissue IHC; UniProt Q15041 tissue specificity). | Report the disagreement with the result. Assess antibody specificity independently before using heart as a reference tissue. |
| Bone-marrow hematopoietic cells are unstained. | HPA records no detected tissue staining, while UniProt reports expression across hematopoietic lineages (HPA tissue IHC; UniProt Q15041 tissue specificity). | Do not use a negative marrow section alone to rule out ARL6IP1. Check technical controls and seek independent evidence if marrow expression matters. |
| Two antibodies yield different cytoplasmic patterns. | HPA's tissue assessment is Uncertain and cautions about recognition of proteins from more than one gene (HPA tissue IHC). | Document each antibody's validation status and compare staining in the same cell populations and controls. Withhold a definitive localization or expression call until the discrepancy is resolved. |
Experiment evidence
IHC & IF images showing ARL6IP1 staining
A08487 · IHCImmunohistochemical staining of human bladder tissue using ARMER antibody at 2 μg/mL.
A08487 · IFImmunofluorescence of ARMER in Mouse Intestine cells with ARMER antibody at 2 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000170540-ARL6IP1/tissue/cerebral+cortex#imid_12091675
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000170540-ARL6IP1/tissue/colon#imid_12091609
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000170540-ARL6IP1/tissue/liver#imid_12091610
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000170540-ARL6IP1/tissue/kidney#imid_12091603
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA045307; green = ARL6IP1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA045307; green = ARL6IP1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ARL6IP1 IHC & IF
🧪Run duodenum first and require staining in its glandular cells (HPA: High in duodenal glandular cells). Run adipose tissue as the negative comparator (HPA: Not detected in adipocytes); on the duodenum slide, use unstained nonglandular areas to judge background, without treating those cells as a validated negative population (HPA: High designation applies to glandular cells).
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 ARL6IP1 in A-431, U-251MG, U2OS, with annotated localisation: Endoplasmic reticulum (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and isotype-matched controls, plus an ARL6IP1 knockout specimen as a biological negative (standard IHC practice). Check endogenous peroxidase in the duodenum section before chromogenic detection, and block endogenous biotin if using biotin-based detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and the effect of antigen retrieval are unreported in the supplied evidence; the selected A08487 bladder IHC caption also leaves the fixative unreported (selected-SKU caption). There is no supplied comparison establishing whether frozen sections or IF are easier for ARL6IP1; IF has a supported ER localization reference, with an antibody-specificity caution (HPA: subcellular summary). For duodenum, assess endogenous peroxidase background during chromogenic IHC (standard IHC practice).
HPA tissue IHC evidence for ARL6IP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene. 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 |
|---|---|---|---|---|
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
| Ovary | Follicle cells | High | Protein (IHC) | HPA → |
| Small intestine | Glandular cells | High | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | High | 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 → |
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ARL6IP1 IHC Tips
These troubleshooting steps center on chromogenic IHC for ARL6IP1 in paraffin sections; IF/ICC is addressed separately.
How should I adjust retrieval when ARL6IP1 staining is weak or morphology deteriorates?
Use citrate pH 6.0 HIER at 95–98 °C for 20 min before chromogenic IHC (page retrieval setting). Keep section thickness, cooling, and detection constant when comparing retrieved sections with a no-retrieval control; excessive retrieval can damage morphology (standard IHC practice). Check antibody concentration and detection on a control section before changing retrieval: the selected bladder tissue image used 2 µg/mL, although its fixative was unreported (selected A08487 caption). If staining remains weak, test another buffer or pH on serial sections as a fallback, recording both signal and background (standard IHC practice).
Could fixation explain inconsistent ARL6IP1 staining across paraffin blocks?
Target-specific fixation sensitivity is unknown: the selected bladder tissue-IHC caption does not report a fixative (selected A08487 caption). Record the fixative, fixation duration, tissue thickness, and processing history for each block before comparing staining (standard IHC practice). For newly collected material, apply one documented fixation schedule across comparison groups and include a consistently processed control; variable processing can change antigen accessibility and morphology (standard IHC practice). Compare serial sections under the same citrate pH 6.0, 95–98 °C, 20 min retrieval setting, then assess whether differences persist without attributing them to an unproven ARL6IP1-specific fixation effect (page retrieval setting; standard IHC practice).
Where should convincing ARL6IP1 staining appear in a tissue section?
Expect a predominantly cytoplasmic, intracellular membrane-associated pattern, consistent with ARL6IP1 enrichment in ER tubules and sheet edges (UniProt Q15041 subcellular annotation). Chromogenic IHC generally cannot resolve individual ER tubules, so score a reproducible cytoplasmic pattern within identifiable cells rather than requiring visible tubules (standard IHC practice). The HPA tissue profile reports cytoplasmic expression, but its antibody staining has uncertain reliability and may include proteins from more than one gene (HPA tissue IHC). Treat isolated nuclear, extracellular, or sharply peripheral staining as a specificity question; compare cellular morphology, a no-primary control, and an independent antibody if available (UniProt Q15041 subcellular annotation; standard IHC practice).
How can epitope position or alternative splicing affect ARL6IP1 IHC?
ARL6IP1 has 3 annotated isoforms, but the supplied evidence does not map the catalog antibody's epitope or establish which isoforms it detects (UniProt Q15041 isoforms; selected A08487 caption). Its annotated topology places residues 1–41 and 87–133 on the cytoplasmic side and 176–203 on the lumenal side (UniProt Q15041 topology). Confirm the immunogen or epitope from antibody documentation before interpreting a negative section as absence of every isoform (standard IHC practice). If an independent antibody recognizes a different mapped region, compare staining on adjacent sections under matched retrieval and detection conditions; disagreement requires investigation, especially given HPA's specificity caution (standard IHC practice; HPA tissue IHC).
How should I investigate ARL6IP1 localisation with multiplex IF?
Use the separate IF/ICC guide for that application; the selected A08487 bladder caption provides tissue-IHC evidence only and does not establish IF conditions (selected A08487 caption). Multiplex ARL6IP1 with a marker for the cell population being assessed, such as a glandular cell marker when examining the HPA-reported duodenal glandular signal (HPA tissue IHC; standard IF practice). Choose a red or far-red fluorophore after checking unstained tissue for autofluorescence, and include single-label controls for bleed-through (standard IF practice). If the antibody epitope is cytoplasmic, optimise permeabilisation for access; if it is lumenal, confirm that membrane access is adequate, because the supplied evidence does not locate this antibody's epitope (UniProt Q15041 topology; standard IF practice).
What should I check when ARL6IP1 DAB staining is diffuse?
Run no-primary and detection-only controls to distinguish antibody-associated staining from endogenous enzyme activity or detection background (standard chromogenic IHC practice). Check the peroxidase block before DAB development, and compare a shorter development time on serial sections if the precipitate obscures cell boundaries (standard chromogenic IHC practice). Titrate the primary around the reported 2 µg/mL used for the selected bladder tissue image, while recognising that its fixative and full detection conditions were not stated (selected A08487 caption). Reduce nonspecific binding with a suitable protein block and adequate washes, then judge improvement by retained intracellular cytoplasmic signal and preserved morphology (UniProt Q15041 subcellular annotation; standard IHC practice).
How should I score ARL6IP1 staining across tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and viable region before scoring, then use the same threshold and imaging settings across sections (standard IHC quantification practice). An H-score combines the percentage of cells at intensity grades 0–3 into a 0–300 result; also report percent positive cells when prevalence matters (standard IHC quantification practice). For spatial comparisons, report positive-cell density per mm² and normalise to viable tissue area or the number of eligible cells, excluding folds and necrosis (standard IHC quantification practice). Record cytoplasmic staining separately from other compartments, and interpret between-tissue differences cautiously because HPA rates its tissue staining uncertain (UniProt Q15041 subcellular annotation; HPA tissue IHC).
How can I distinguish a true ARL6IP1 signal from artefact?
A plausible signal is intracellular and predominantly cytoplasmic, matching the annotated ER membrane localisation rather than isolated nuclear or extracellular deposit (UniProt Q15041 subcellular annotation; standard IHC practice). Check whether staining follows intact cells instead of section edges, folds, necrotic areas, or endogenous peroxidase-rich regions, and compare no-primary controls (standard chromogenic IHC practice). Cell identity matters: HPA reports high staining in duodenal glandular cells, while its overall tissue-IHC reliability is uncertain because the antibody may recognize proteins from multiple genes (HPA tissue IHC). Resolve unexpected patterns with an independent antibody or other orthogonal evidence before calling them ARL6IP1-specific, particularly when a tissue result conflicts with the expression record (HPA tissue IHC; UniProt Q15041 tissue specificity; standard IHC practice).
Boster reagents
Best ARL6IP1 / ADP-ribosylation factor-like protein 6-interacting protein 1 IHC Antibodies
A08487 has IHC-P and IF applications for human and mouse samples (catalog applications/reactivity), with IHC shown in human bladder tissue and IF shown in mouse intestine cells (image captions).
Real IHC data
Anti-ARMER ARL6IP1 Antibody
Cat # A08487
A08487 will render with an IHC image of human bladder tissue stained at 2 μg/mL (IHC image caption). Its IF image shows mouse intestine cells stained at 2 μg/mL (IF image caption).
Which to pick: For tissue IHC, choose A08487: IHC-P is listed (catalog applications), and its IHC image shows human bladder tissue; the caption does not report the fixative or section processing (IHC image caption). For IF/ICC, A08487 lists IF and shows staining in mouse intestine cells (catalog applications; IF image caption). For human and mouse work, A08487 lists both species and has a rabbit host; its clonality is unreported (catalog reactivity/host/clone).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q15041 (AR6P1_HUMAN, ADP-ribosylation factor-like protein 6-interacting protein 1).
- Human Protein Atlas. ARL6IP1 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. ARL6IP1 subcellular location (ICC-IF): Localized to the endoplasmic reticulum. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. ARL6IP1 antibody validation summary (1 antibodies).
- Identification and functional validation of a four-gene signature associated with radiotherapy resistance in oral squamous cell carcinoma. Translational oncology 2026 — PMC13383874.
- Liver X receptor-agonist treatment rescues degeneration in a Drosophila model of hereditary spastic paraplegia. Acta neuropathologica communications 2022 — PMC8961908.
- PubMed PMID:7584044 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15616553 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ARL6IP1 (ADP-ribosylation factor-like protein 6-interacting protein 1). Expected localisation Cytoplasmic tissue staining (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: A08487 at 2 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Duodenum (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: human bladder tissue; fixation not specified (datasheet A08487); 2. Fixation: Image fixative and duration unreported (datasheet A08487); 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-ARL6IP1, 2 μg/mL (datasheet A08487); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining may detect proteins from more than one gene (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unreported (UniProt); Regulation: Highest in early myeloid progenitors (UniProt). Evidence: UniProt Q15041, HPA tissue IHC · Uncertain, HPA ICC-IF · endoplasmic reticulum. Caution: Based on antibodies targeting…, Datasheet A08487, PMC13383874. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ARL6IP1/immunohistochemistry-ihc-guide.