This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
ARL2 / ADP-ribosylation factor-like protein 2 · IHC design guide
Design Immunohistochemistry for ARL2
Plan ARL2 staining in paraffin sections using the nuclear and cytoplasmic tissue pattern and high expression in colon glandular cells (HPA tissue IHC). The catalog antibody has a documented IHC range of 2–5 μg/ml (datasheet A04155-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ARL2 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 | Nuclear and cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Glandular cells show nuclear and cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04155-1) | |
| Positive control | Colon+4 more · see all | |
| Negative control | Liver |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Liver cholangiocytes may show no detectable staining (HPA tissue IHC) | |
| Regulation | Low tissue specificity; no inducer established (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended ARL2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with two published ARL2 paraffin-section protocols (PMC7816291; PMC5975491).
| Sample | Paraffin-embedded human cervical cancer tissue; fixative not specified (datasheet A04155-1) |
| Fixation | Image fixative and duration unreported (datasheet A04155-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A04155-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04155-1) |
| Primary antibody | Rabbit anti-ARL2, 2-5 μg/ml (datasheet A04155-1) |
| Primary incubation | Overnight at 4 °C (datasheet A04155-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04155-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ARL2-positive staining in glandular cells of colon (HPA tissue IHC: High). HPA tissue profile: General nuclear and cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 (datasheet A04155-1); citrate at pH 6.0 is a published alternative (PMC5975491).
Section 2
What Is the Expected ARL2 Staining Pattern?
ARL2 should show nuclear and cytoplasmic staining in several cell types, including colon and epididymal glandular cells, fallopian tube ciliated cells, and pancreatic exocrine glandular cells (HPA tissue IHC: Supported; High). Centrosome association and mitochondrial localization are also reported (UniProt P36404 subcellular location). ARL2 has no transmembrane segment, so a solely membrane-bound pattern would need scrutiny (UniProt P36404 topology).
| Nuclear and cytoplasmic chromogen appears in glandular or ciliated cells, with stronger signal than nearby unstained areas. | This fits the general tissue profile; HPA reports High staining in colon glandular cells and fallopian tube ciliated cells (HPA tissue IHC: Supported; High). Score the named cell population and compartment separately, since tissue-wide intensity can hide a restricted pattern (general IHC practice). |
| Signal is confined to an unexpected compartment, such as a continuous cell outline, with no convincing nuclear or cytoplasmic staining. | Treat an isolated outline as questionable: HPA describes general nuclear and cytoplasmic tissue expression, while UniProt reports no transmembrane segment (HPA tissue IHC; UniProt P36404 topology). Check morphology and controls before calling it ARL2; neither source makes every membrane-adjacent signal an artifact. |
| Cholangiocytes stain strongly while an expected positive cell population on the run is weak or blank. | HPA reports ARL2 as Not detected in liver cholangiocytes, so this discordance raises concern for cross-reactivity or endogenous detection activity (HPA tissue IHC: cholangiocytes, Not detected; general IHC practice). That reference finding is a comparator, not proof that every cholangiocyte must be negative. |
| Diffuse chromogen covers tissue, lumina, or blank slide areas and obscures cell boundaries. | This distribution cannot be scored as the cell-associated nuclear and cytoplasmic profile described by HPA (HPA tissue IHC). Non-target-specific possibilities include incomplete blocking, excessive detection signal, or residual reagent; inspect a no-primary control before attributing the color to ARL2 (general IHC practice). |
| A known positive tissue shows no interpretable signal in the named cell population. | Absence in a reported High population, such as colon glandular cells, conflicts with the HPA reference pattern (HPA tissue IHC: colon glandular cells, High). First assess section quality and detection controls; a failed staining run cannot establish biological absence (general IHC practice). |
💡Expected ARL2 appearanceCall a result positive when cell-associated nuclear and cytoplasmic staining is clear in a reported High population, such as colon glandular cells; diffuse acellular color alone is a false-positive pattern for interpretation (HPA tissue IHC: Supported; colon glandular cells, High; general IHC practice).
| Tissue and cell population | HPA reports Low tissue specificity at the RNA level and a general nuclear and cytoplasmic IHC profile, yet staining levels differ by cell population (HPA tissue IHC). Use its named High populations as positive comparators and its Not detected cholangiocytes as a cautious negative comparator (HPA tissue IHC). |
| Compartment evidence | HPA tissue IHC supports nuclear and cytoplasmic staining; UniProt lists mitochondria and centrosomes but also says ARL2 was not detected in the nucleus in its location note (HPA tissue IHC; UniProt P36404 subcellular location). Record observed compartments without forcing these annotations into a single-cell interpretation. |
| Antibody evidence | The listed antibody HPA044610 has Supported IHC and Supported ICC status, and HPA calls the tissue IHC reliability Supported based on staining and RNA consistency (HPA antibodies; HPA tissue IHC). This supports pattern comparison, but does not provide an epitope, dilution, retrieval condition, or fixation-sensitivity claim. |
| Isoforms and processing | UniProt lists two ARL2 isoforms, no signal peptide or propeptide, and a chain annotated from residues 2–184 (UniProt P36404 isoforms and processing). Without a supplied antibody epitope, these annotations cannot predict which isoform is detected or justify a distinct staining pattern. |
| IF/ICC Q&A: What localization should be expected? | HPA reports mainly nucleoplasm, basal body, and flagellar centriole, with additional nucleolar and mitochondrial locations, among others (HPA subcellular ICC-IF). These imaging annotations can guide interpretation of an IF/ICC result; they do not replace the tissue IHC reference pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Colon glandular cells are blank. | The result disagrees with HPA's High reference staining; the cause cannot be assigned from that comparison alone (HPA tissue IHC: colon glandular cells, High). | Check section integrity, a same-run positive control, and detection performance before scoring the specimen; follow the antibody's documented IHC-P conditions where available (general IHC practice). |
| All cells and blank areas look brown. | Widespread acellular color is inconsistent with interpretable cell-associated staining (HPA tissue IHC; general IHC practice). | Review a no-primary control, blocking and wash steps, and detection exposure; then reassess nuclear and cytoplasmic localization in intact cells (general IHC practice). |
| Only cell outlines stain. | A solely outlined pattern is discordant with HPA's general nuclear and cytoplasmic profile and UniProt's lack of a transmembrane segment (HPA tissue IHC; UniProt P36404 topology). | Compare with positive tissue and controls, inspect whether the signal follows cells or deposits, and withhold a compartment call until morphology supports it (general IHC practice). |
| Cholangiocytes appear strongly positive. | HPA reports Not detected staining in this population; cross-reactivity or endogenous detection activity is a possible explanation, not an established diagnosis (HPA tissue IHC; general IHC practice). | Check a no-primary control and compare the same run with a reported High population; record the discordance if controls pass (general IHC practice; HPA tissue IHC). |
| Nuclear signal seems at odds with the UniProt note. | HPA describes nuclear tissue staining and supported nucleoplasmic ICC-IF localization, while a UniProt location note says ARL2 was not detected in the nucleus (HPA tissue IHC; HPA subcellular ICC-IF; UniProt P36404 subcellular location). | Report the observed compartment and the source discrepancy; judge the IHC slide against HPA tissue images and staining controls (HPA tissue IHC; general IHC practice). |
| Signal is weak in a low-staining population. | HPA reports Low staining in hippocampal neuronal, smooth muscle, and bone marrow hematopoietic cells, so weak signal alone need not mean the run failed (HPA tissue IHC: Low). | Compare with a reported High population on a valid run before changing staining conditions; score the relevant cell type and avoid treating a Low reference as an absolute negative (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing ARL2 staining
A04155-1 · IHCIHC analysis of ARL2 using anti-ARL2 antibody (A04155-1). ARL2 was detected in a paraffin-embedded section of human cervical cancer tissue.
A04155-1 · IFIF analysis of ARL2 using anti-ARL2 antibody (A04155-1). ARL2 was detected in an immunocytochemical section of HELA cells. Enzyme antigen re
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000213465-ARL2/tissue/cerebral+cortex#imid_12184425
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000213465-ARL2/tissue/colon#imid_12184335
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000213465-ARL2/tissue/liver#imid_12184352
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000213465-ARL2/tissue/kidney#imid_12184345
HPA · ICC-IF U2OSHuman Protein Atlas immunofluorescence, U2OS, antibody HPA044610; green = ARL2, blue = nucleus, red = microtubules.
HPA · ICC-IF U2OSHuman Protein Atlas immunofluorescence, U2OS, antibody HPA044610; green = ARL2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ARL2 IHC & IF
🧪Run colon first: glandular cells should stain at the High level (HPA: Colon, glandular cells, High). Run liver cholangiocytes as the negative tissue (HPA: Liver, cholangiocytes, Not detected); use cells outside the colon glands as internal background comparators only after confirming their staining is nonspecific, because the HPA colon row identifies no internal-negative cell type (HPA: Colon, glandular cells, High).
Positive control tissue: Colon (Glandular cells, HPA High)
Negative control tissue: Liver (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ARL2 in U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), Sperm, with annotated localisation: Nucleoplasm (supported), Basal body (approved), Flagellar centriole (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, matched rabbit IgG isotype control with clonality matched where applicable, and ARL2 knockout material or cognate peptide competition if available (caption: rabbit anti-ARL2; standard IHC controls). For colon, quench endogenous peroxidase and inspect gland lumens for DAB background (caption: HRP/DAB detection; standard chromogenic IHC practice).
⚠️Feasibility: No supplied source reports a target-specific fixation window or fixation effect; the selected A04155-1 paraffin-section caption leaves the fixative unreported (caption: fixative not stated). The caption uses heat-mediated EDTA retrieval at pH 8.0, but does not establish that retrieval is required; the supplied evidence also does not establish whether frozen sections or IF are easier (caption: EDTA retrieval; HPA: ICC-IF images). In colon glands, assess any luminal DAB deposit separately from cellular staining (HPA: Colon, glandular cells, High; standard chromogenic IHC practice).
HPA tissue IHC evidence for ARL2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — High 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 |
|---|---|---|---|---|
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Lung | Alveolar cells type I | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ARL2 IHC Tips
Use compartment, cell type and matched controls to troubleshoot ARL2 staining in paraffin sections; interpret IF as supporting spatial evidence.
How should I adjust retrieval when ARL2 staining is weak or uneven?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A04155-1). The selected tissue image used this retrieval before staining with 2 μg/ml catalog antibody overnight at 4°C (caption A04155-1). If signal is weak, compare retrieval heating and cooling conditions on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Check whether tissue edges stain more strongly than the interior, since uneven processing can mimic a retrieval problem (standard IHC practice). If the recommended condition remains weak, test an alternative retrieval buffer as a documented fallback and compare morphology alongside signal (standard IHC practice).
Could fixation explain inconsistent ARL2 staining across paraffin blocks?
Target-specific ARL2 fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative or fixation duration (caption A04155-1). Record those details for each block and compare sections processed with the same retrieval, antibody incubation and chromogenic detection settings (standard IHC practice). Excessive or uneven fixation can alter epitope accessibility in IHC generally, so assess staining together with tissue morphology rather than assigning an ARL2-specific effect (standard IHC practice). If blocks differ, repeat staining with matched control tissue in the same run and document whether the discrepancy follows the block or the staining run (standard IHC practice).
Which ARL2 staining patterns should I expect in chromogenic tissue sections?
Assess nuclear and cytoplasmic staining because the tissue IHC profile reports general expression in both compartments (HPA tissue IHC). ARL2 is also associated with mitochondrial intermembrane space and centrosomes, although routine chromogenic sections may not resolve those structures individually (UniProt P36404 subcellular location; standard IHC practice). High staining in fallopian-tube ciliated-cell rootlets offers a more spatially specific comparison when that tissue is available (HPA tissue IHC). Score compartment and cell type separately, then compare the pattern with a matched negative control and preserved morphology (standard IHC practice). A nuclear signal alone should not be dismissed solely on the basis of the UniProt note reporting no nuclear detection (HPA tissue IHC; UniProt P36404 subcellular location).
Can this IHC antibody distinguish ARL2 isoforms or modified epitopes?
Do not assign isoform specificity from tissue staining alone: ARL2 has 2 annotated isoforms, while the supplied antibody caption does not map its epitope (UniProt P36404 isoforms; caption A04155-1). ARL2 is 184 amino acids long and has an annotated phosphoserine at residue 45, but the supplied evidence does not establish whether that residue lies within the antibody epitope (UniProt P36404 length and modified residues; caption A04155-1). Its lack of a transmembrane segment does not identify an exposed antibody epitope after tissue processing (UniProt P36404 topology; standard IHC practice). For an isoform or modification claim, obtain epitope mapping and validate the proposed specificity with appropriately characterized material (standard IHC practice).
How can IF help resolve an ambiguous ARL2 IHC pattern?
Use IF on a matched specimen to examine compartment overlap when chromogenic ARL2 staining is difficult to assign to individual structures (standard IHC/IF practice). Multiplex ARL2 with a marker for the expected cell population, such as a ciliated-cell marker when investigating the HPA rootlet pattern, and validate each channel separately (HPA tissue IHC; standard IF practice). Choose a fluorophore channel with low tissue autofluorescence and check an unstained specimen before interpreting faint puncta (standard IF practice). ARL2 lacks a transmembrane segment but includes a mitochondrial intermembrane-space pool; because this antibody's epitope is unmapped, optimize permeabilisation for the compartment being tested rather than assuming an accessible membrane side (UniProt P36404 topology and subcellular location; caption A04155-1).
What should I check when ARL2 DAB staining is widespread?
Check a section processed without primary antibody to identify detection-system background, including endogenous peroxidase activity (standard IHC practice). The selected image used a peroxidase-conjugated secondary antibody and DAB, so a peroxidase block and consistent DAB development are relevant workflow controls (caption A04155-1; standard IHC practice). The documented protocol blocked with 10% goat serum and incubated the catalog antibody at 2 μg/ml overnight at 4°C (caption A04155-1). If background persists, compare antibody concentration and washing conditions on adjacent sections while keeping retrieval and development matched (standard IHC practice). Treat diffuse staining cautiously because ARL2 can appear in both nuclear and cytoplasmic compartments (HPA tissue IHC).
How should I quantify ARL2 across heterogeneous tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population, tissue region and nuclear versus cytoplasmic scoring rules before comparing slides (HPA tissue IHC; standard IHC practice). For cell-level staining, report the percentage of positive cells and an H-score using intensity grades 0–3; apply the same thresholds across specimens (standard IHC practice). For a spatial feature such as ciliary rootlets, consider positive structures per mm² of evaluable tissue instead of treating every brown pixel as a cell (HPA tissue IHC; standard IHC practice). Normalize counts to the relevant cell population or analyzable area, and exclude folds, necrosis and poorly preserved regions consistently (standard IHC practice). Include matched controls to monitor run-level shifts in chromogen intensity (standard IHC practice).
How do I separate true ARL2 signal from tissue artefact?
Require staining in intact cells with a plausible compartment and cell-type pattern, rather than accepting DAB deposits at section edges or within necrosis (HPA tissue IHC; standard IHC practice). Nuclear and cytoplasmic signals are plausible in tissue, while ciliary-rootlet enrichment has been reported in fallopian-tube ciliated cells (HPA tissue IHC). Compare candidate positives with a no-primary control to identify endogenous enzyme or secondary-reagent signal, particularly when pigment resembles the chromogen (standard IHC practice). A centrosomal or mitochondrial assignment needs spatial resolution or corroborating markers beyond routine DAB morphology (UniProt P36404 subcellular location; standard IHC practice). Use the reported high glandular-cell staining and undetected liver cholangiocyte staining as contextual references, not universal thresholds (HPA tissue IHC).
Boster reagents
Best ARL2 / ADP-ribosylation factor-like protein 2 IHC Antibodies
The IHC-validated antibody A04155-1 has human cervical cancer paraffin-section IHC and HeLa-cell IF/ICC images (catalog captions); listed reactivity covers human, mouse and rat (catalog reactivity).
Real IHC data
Anti-ARL2 Antibody ®
Cat # A04155-1
A04155-1 will render with an IHC image from human cervical cancer paraffin sections (catalog IHC caption). Its IF/ICC image shows HeLa cells (catalog IF caption), and its listed reactivity is human, mouse and rat (catalog reactivity).
Which to pick: For tissue IHC, choose A04155-1: its own caption documents human cervical cancer paraffin sections, EDTA pH 8.0 retrieval and 2 μg/ml primary antibody (catalog IHC caption); the fixative is unreported (catalog IHC caption). For IF/ICC, the same SKU has a HeLa-cell image using 5 μg/ml primary antibody (catalog IF caption). For cross-species planning, A04155-1 lists human, mouse and rat reactivity (catalog reactivity), while its supplied IHC image documents human tissue (catalog IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P36404 (ARL2_HUMAN, ADP-ribosylation factor-like protein 2).
- Human Protein Atlas. ARL2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. ARL2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm, basal body and flagellar centriole. In addition localized to the nucleoli, mitochondria, equatorial segment and mid piece..
- Human Protein Atlas. ARL2 antibody validation summary (1 antibodies).
- Regulation of ARL2 in colorectal cancer cell proliferation and tumorigenicity, and its negative association with AXL. Oncology letters 2021 — PMC7816291.
- ARL2 overexpression inhibits glioma proliferation and tumorigenicity via down-regulating AXL. BMC cancer 2018 — PMC5975491.
- ARL2 is required for homologous recombination repair and colon cancer stem cell survival. FEBS open bio 2022 — PMC9340879.
- ADP-Ribosylation Factor-Like 2 (ARL2) regulates cilia stability and development of outer segments in rod photoreceptor neurons. Scientific reports 2018 — PMC6240099.
- PubMed PMID:8415637 — UniProt-cited evidence.
- PubMed PMID:15146197 — UniProt-cited evidence.
- PubMed PMID:16554811 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ARL2 (ADP-ribosylation factor-like protein 2). Expected localisation Nuclear and cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04155-1). Validated IHC antibody: A04155-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Colon (Glandular cells); negative Liver. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human cervical cancer tissue; fixative not specified (datasheet A04155-1); 2. Fixation: Image fixative and duration unreported (datasheet A04155-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A04155-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04155-1); 8. Primary antibody: Rabbit anti-ARL2, 2-5 μg/ml (datasheet A04155-1); 9. Primary incubation: Overnight at 4 °C (datasheet A04155-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Liver cholangiocytes may show no detectable staining (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unknown (UniProt); Regulation: Low tissue specificity; no inducer established (HPA tissue IHC). Evidence: UniProt P36404, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm, basal body and flagellar…, Datasheet A04155-1, PMC7816291, PMC5975491. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ARL2/immunohistochemistry-ihc-guide.