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ARL13B / ADP-ribosylation factor-like protein 13B · IHC design guide
Design Immunohistochemistry for ARL13B
This guide plans paraffin-section ARL13B IHC around the cytoplasmic tissue staining reported by HPA and the ciliary membrane location annotated by UniProt (HPA tissue IHC; UniProt). Adrenal gland and gallbladder glandular cells provide reported high-staining references (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ARL13B 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 | Cytoplasm in tissue; ciliary membrane molecularly (HPA tissue IHC; UniProt) | |
| Staining pattern | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04279-1) | |
| Positive control | Adrenal gland+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 sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining has medium concordance with RNA (HPA tissue IHC) | |
| Regulation | Expressed in the developing brain (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended ARL13B IHC & IF Protocols
The catalog antibody's IHC-P protocol (datasheet A04279-1) is paired with two published chromogenic ARL13B tissue protocols (PMC5442446; PMC6826833).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A04279-1) |
| Fixation | Image fixative and duration unreported (datasheet A04279-1); 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 A04279-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04279-1) |
| Primary antibody | Rabbit anti-ARL13B, 2-5 μg/ml (datasheet A04279-1) |
| Primary incubation | Overnight at 4 °C (datasheet A04279-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04279-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ARL13B-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A04279-1). Use citrate pH 6.0 when reproducing the breast tissue protocol (PMC6826833).
Section 2
What Is the Expected ARL13B Staining Pattern?
ARL13B associates with the cilium membrane and cytoskeleton; it has no transmembrane segment (UniProt Q3SXY8 localization and topology). In tissue IHC, expect predominantly cytoplasmic staining, with stronger signal in some glandular and alveolar cells (HPA: tissue IHC). HPA rates its tissue staining Approved, with medium consistency between staining and RNA expression (HPA: tissue IHC reliability).
| Cytoplasmic stain in adrenal or gallbladder glandular cells, or lung alveolar type I cells. | These are plausible positive patterns: HPA records High staining in each listed cell population (HPA: tissue IHC). A discrete ciliary signal can also fit ARL13B localization, but a cilium need not be visibly resolved in a chromogenic tissue section (UniProt Q3SXY8 localization; general IHC practice). |
| Stain confined to nuclei, with no matching cytoplasmic or ciliary signal. | Treat an isolated nuclear pattern as suspect because the supplied localization records place ARL13B at cilia, microtubules and the cytoskeleton, while tissue IHC is predominantly cytoplasmic (UniProt Q3SXY8 localization; HPA: tissue IHC). Review morphology and controls before scoring such cells as positive (general IHC practice). |
| A strong signal appears in an unexpected cell population. | Compare the exact cell type with the tissue record: HPA lists Low staining in bronchial respiratory epithelium and liver cholangiocytes, for example (HPA: tissue IHC). An unexpectedly strong pattern can reflect cross-reactivity or endogenous detection activity; the HPA list contains no declared negative cell population that would prove absence (HPA: tissue IHC; general IHC practice). |
| Colour spreads across stroma, lumina or much of the section without cell boundaries. | Diffuse deposit is difficult to assign to the cytoplasmic pattern HPA describes (HPA: tissue IHC). Check the no-primary control and inspect whether signal tracks tissue structures; background may arise from nonspecific antibody binding, endogenous detection activity or excessive chromogen development (general IHC practice). |
| No signal appears in a section expected to contain adrenal or gallbladder glandular cells. | Those cells are reported as High in HPA tissue IHC, so their absence warrants a workflow check (HPA: tissue IHC). Confirm that the expected cells are present and preserved, then review the IHC-validated antibody, detection reagents and control section (general IHC practice). HPA's Approved rating does not guarantee every specimen will stain (HPA: tissue IHC reliability). |
💡Expected ARL13B appearanceCall a section positive when identifiable cells show cytoplasmic signal, particularly High signal in the HPA-listed adrenal or gallbladder glandular cells or lung alveolar type I cells; isolated nuclear colour or structure-free haze is suspect (HPA: tissue IHC; UniProt Q3SXY8 localization; general IHC practice).
| Cell type and tissue | HPA reports High staining in adrenal and gallbladder glandular cells and lung alveolar type I cells, but Medium staining in endometrial glandular cells and adipocytes (HPA: tissue IHC). Select and score controls by cell type, not by whole-section darkness (general IHC practice). |
| Compartment and section resolution | UniProt places ARL13B at ciliary membranes and the cytoskeleton; HPA describes tissue IHC as cytoplasmic in most tissues (UniProt Q3SXY8 localization; HPA: tissue IHC). A visible cilium is supportive when resolved, while cytoplasmic staining remains compatible with the reported tissue pattern (general IHC practice). |
| Antibody evidence | HPA048926 is listed as Approved for IHC and Supported for ICC, while overall tissue staining has medium consistency with RNA expression (HPA: antibody validation; HPA: tissue IHC reliability). Interpret a new specimen against its cell morphology and controls rather than treating validation as a specimen-level guarantee (general IHC practice). |
| Isoforms and target features | UniProt lists 3 isoforms, no transmembrane segment, and cilium-membrane association via palmitoylation (UniProt Q3SXY8 isoforms, topology and localization). The payload gives no antibody epitope, so it cannot establish which isoforms the IHC-validated antibody detects or predict an antigen-retrieval result. |
| What should IF/ICC show? | HPA supports localization at microtubules, primary cilia and the primary cilium transition zone in ICC-IF (HPA: subcellular ICC-IF). UniProt describes proximal ciliary membranes while excluding the transition zone (UniProt Q3SXY8 localization). Treat this compartment difference as a reason to check the assay context, not as a required chromogenic IHC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in an expected positive cell population. | The target cells may be absent from the section, or an IHC workflow step may have failed (general IHC practice). | Confirm glandular or alveolar cell identity, then check a positive control and the IHC-validated antibody and detection steps; HPA reports High staining in the corresponding listed cells (HPA: tissue IHC; general IHC practice). |
| Only nuclear staining is visible. | That pattern conflicts with the supplied cytoplasmic and ciliary localization evidence (HPA: tissue IHC; UniProt Q3SXY8 localization). | Review the no-primary control, cell morphology and antibody specificity before calling nuclei positive (general IHC practice). |
| Strong staining appears in a cell type reported as Low. | Nonspecific binding or endogenous detection activity is possible; the HPA level is a reference pattern, not proof of the cause (HPA: tissue IHC; general IHC practice). | Compare matched controls and score the named cell type separately; bronchial respiratory epithelium and liver cholangiocytes are Low in HPA (HPA: tissue IHC; general IHC practice). |
| Diffuse colour obscures cellular boundaries. | Background from nonspecific binding, endogenous activity or chromogen development can obscure interpretation (general IHC practice). | Inspect a no-primary control and review blocking, antibody dilution and development time using the chosen IHC workflow (general IHC practice). |
| Cilia are not individually visible, but cytoplasmic staining is present. | A chromogenic section may not resolve a small ciliary compartment even though ARL13B localizes there (UniProt Q3SXY8 localization; general IHC practice). | Score the cell-level cytoplasmic pattern against HPA's tissue IHC observations; avoid making visible cilia a universal acceptance requirement (HPA: tissue IHC; general IHC practice). |
| An antigen-retrieval change alters the signal. | The supplied sources establish no ARL13B-specific fixation or retrieval sensitivity. | Compare the changed condition with a consistent positive control and no-primary control; report the observed change without attributing it to ARL13B epitope masking (general IHC practice). |
Experiment evidence
IHC & IF images showing ARL13B staining
A04279-1 · IHCIHC analysis of ARL13B using anti-ARL13B antibody (A04279-1). ARL13B was detected in a paraffin-embedded section of human breast cancer tiss
A04279-1 · IHCIHC analysis of ARL13B using anti-ARL13B antibody (A04279-1). ARL13B was detected in a paraffin-embedded section of human colorectal adenoca
A04279-1 · IFIF analysis of ARL13B using anti-ARL13B antibody (A04279-1). ARL13B was detected in an immunocytochemical section of U20S cells. Enzyme anti
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000169379-ARL13B/tissue/cerebral+cortex#imid_14081026
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000169379-ARL13B/tissue/colon#imid_14080960
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000169379-ARL13B/tissue/liver#imid_14080969
HPA · GallbladderHuman Protein Atlas IHC, Gallbladder. Source: https://www.proteinatlas.org/ENSG00000169379-ARL13B/tissue/gallbladder#imid_14080945
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA048926; green = ARL13B, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA048926; green = ARL13B, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ARL13B IHC & IF
🧪Run gallbladder first; its glandular cells must show ARL13B staining (HPA: High in gallbladder glandular cells). HPA detects ARL13B in all 45 scored tissues, so use no-primary and isotype controls for negative staining; neighboring non-glandular cells may serve as internal background comparators only if they show no discrete ciliary signal, since no negative cell population is established for this slide (HPA: no negative tissue rows; standard IHC scoring practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: ARL13B 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 ARL13B in A-431, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), Sperm, NIH 3T3, with annotated localisation: Microtubules (supported), Primary cilium (supported), Primary cilium transition zone (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, an isotype control matched to the primary antibody’s rabbit host, immunoglobulin class and clonality, and an ARL13B knockout or validated peptide-block control (caption: rabbit primary; standard IHC control practice). Quench endogenous peroxidase and check gallbladder bile pigment before interpreting brown DAB signal (standard chromogenic IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A04279-1 paraffin-section caption does not state a fixative (caption: fixative unreported). Heat-mediated retrieval in EDTA at pH 8.0 is documented for that IHC example, but its necessity and frozen-section performance are unreported (caption: EDTA pH 8.0 retrieval). IF/ICC may make a discrete ciliary signal easier to resolve, consistent with the reported primary-cilium localization; gallbladder bile pigment can complicate chromogenic interpretation (HPA: supported primary-cilium localization; standard microscopy practice).
HPA tissue IHC evidence for ARL13B
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 → |
| Endometrium | Glandular cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Lung | Alveolar cells type I | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: ARL13B is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced ARL13B IHC Tips
Troubleshoot ARL13B paraffin-section IHC by checking retrieval, tissue quality, staining compartment and controls before interpreting a chromogenic signal.
Which retrieval condition should I use for ARL13B in paraffin sections?
Begin with heat-mediated EDTA retrieval at pH 8.0 for paraffin-section IHC with A04279-1 (datasheet A04279-1). The selected image also used 2 μg/ml primary antibody overnight at 4°C, so hold antibody concentration and detection conditions steady while assessing retrieval (selected SKU IHC caption; standard IHC practice). If staining is weak, compare retrieval heating times on adjacent sections and inspect morphology before changing buffer chemistry (standard IHC practice). Score a tissue control alongside each condition, because stronger diffuse DAB does not establish better detection of ciliary ARL13B (UniProt Q3SXY8 localisation; standard IHC practice).
Could fixation explain weak or uneven ARL13B staining?
The selected paraffin-section image does not state its fixative, so ARL13B-specific fixation sensitivity is unknown (selected SKU IHC caption). Record the specimen's fixative and fixation interval, then compare sections with similar processing before attributing a staining difference to ARL13B biology (standard IHC practice). Examine folds, damaged edges and poorly preserved areas separately, since these can make chromogenic signal uneven across a section (standard IHC practice). Keep EDTA retrieval at pH 8.0 and the primary-antibody condition constant during that comparison; neither tissue staining patterns nor ciliary membrane association establishes a preferred fixation condition (datasheet A04279-1; selected SKU IHC caption; HPA tissue IHC; UniProt Q3SXY8 localisation).
What staining pattern supports ARL13B localisation in tissue IHC?
ARL13B associates with the cilium membrane through palmitoylation and also has reported cytoskeletal localisation (UniProt Q3SXY8 localisation). Its proximal ciliary membrane localisation excludes the transition zone in the cited UniProt description, while cell imaging lists the primary cilium, transition zone and microtubules as supported sites (UniProt Q3SXY8 localisation; HPA subcellular). In paraffin-section IHC, assess discrete projection-associated signal where morphology permits, while recognizing that section angle and chromogenic resolution can obscure a short cilium (standard IHC practice). Diffuse cytoplasmic staining alone is less spatially specific, although cytoplasmic expression across most tissues is reported in tissue IHC (HPA tissue IHC; standard IHC practice).
Can this antibody distinguish ARL13B isoforms or reveal its membrane-facing epitope?
The record lists 3 ARL13B isoforms, but the supplied IHC caption gives no antibody epitope or isoform coverage (UniProt Q3SXY8 isoforms; selected SKU IHC caption). Do not assign a chromogenic pattern to one isoform without an epitope map and isoform-specific validation (standard IHC practice). ARL13B has no annotated transmembrane segment and associates with the cilium membrane through palmitoylation; these facts do not establish which side contains this antibody's epitope (UniProt Q3SXY8 topology and localisation). Compare appropriate positive tissue and negative controls under identical retrieval and detection conditions before treating a compartment difference as evidence of epitope accessibility (HPA tissue IHC; standard IHC practice).
How can IF help resolve an ambiguous ARL13B IHC pattern?
Use IF on a separate section to compare ARL13B with a validated marker for the cell population being scored in IHC; the catalog image itself documents chromogenic paraffin-section IHC (selected SKU IHC caption; standard IF practice). Choose spectrally separated fluorophores and inspect an unstained section for tissue autofluorescence before judging overlap (standard IF practice). Because the antibody epitope's membrane-facing side is unspecified, compare gently permeabilised and nonpermeabilised preparations if access to an intracellular epitope is in question (selected SKU IHC caption; UniProt Q3SXY8 topology; standard IF practice). Interpret apparent ciliary overlap against the reported cilium and microtubule localisation, then return to the matched IHC morphology when scoring tissue (UniProt Q3SXY8 localisation; HPA subcellular; standard IHC practice).
How should I troubleshoot diffuse or patchy DAB background?
The selected IHC image used 10% goat-serum block, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-linked secondary with DAB detection (selected SKU IHC caption). First compare a no-primary control and inspect whether background follows edges, folds or damaged tissue rather than the expected cellular structures (standard IHC practice; UniProt Q3SXY8 localisation). Include a peroxidase block in the general chromogenic workflow and verify wash quality and secondary-antibody specificity before increasing primary-antibody dilution (standard IHC practice). If diffuse signal persists, titrate the primary on comparable sections while retaining EDTA at pH 8.0 and a positive tissue control (datasheet A04279-1; HPA tissue IHC; standard IHC practice).
How should ARL13B staining be scored across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored compartment and cell population before counting, because ARL13B has reported ciliary and cytoskeletal locations while tissue IHC describes cytoplasmic expression in most tissues (UniProt Q3SXY8 localisation; HPA tissue IHC). For broad chromogenic staining, report the percentage of positive eligible cells and an H-score based on intensity and proportion; apply one threshold across specimens (standard IHC practice). For resolvable discrete ciliary profiles, report positive profiles per mm² of evaluable tissue and normalize to eligible cells or tissue area (standard IHC practice). Exclude folds, necrosis and edges consistently, and record retrieval and detection conditions alongside scores (standard IHC practice; datasheet A04279-1).
When is an ARL13B-positive IHC result convincing?
A convincing result is reproducible in the expected cellular context and includes spatial evidence compatible with ciliary or cytoskeletal ARL13B, where section resolution allows (UniProt Q3SXY8 localisation; HPA subcellular; standard IHC practice). Tissue IHC reports high staining in adrenal and gallbladder glandular cells and lung alveolar type I cells, offering documented contexts for comparison (HPA tissue IHC). Treat isolated DAB at section edges, necrotic areas or unexpected compartments cautiously, and check a no-primary control for endogenous enzyme or detection artefact (standard IHC practice). Cytoplasmic staining may be genuine, but intensity alone cannot establish ciliary localisation or isoform identity (HPA tissue IHC; UniProt Q3SXY8 isoforms; standard IHC practice).
Boster reagents
Best ARL13B / ADP-ribosylation factor-like protein 13B IHC Antibodies
A04279-1 has IHC images from human paraffin sections and an IF image from U20S cells; its listed reactivity is Human, Mouse and Rat (catalog image captions; catalog reactivity).
Real IHC data
Anti-ARL13B Antibody ®
Cat # A04279-1
A04279-1 is the only SKU shown, with IHC images from paraffin sections of human breast cancer, colorectal adenocarcinoma, gastric carcinoma and testicular cancer (catalog IHC captions). Its IF image shows U20S cells, and its application list includes IHC, ICC and IF (catalog IF caption; catalog applications).
Which to pick: For tissue IHC, choose A04279-1: its paraffin-section caption documents EDTA retrieval at pH 8.0, a 10% goat serum block and 2 μg/ml primary antibody overnight at 4°C (catalog IHC caption). For IF/ICC, the same SKU has an IF image in U20S cells at 5 μg/ml (catalog IF caption). For cross-species work, A04279-1 lists Human, Mouse and Rat reactivity, while its IHC captions show human sections; the fixative is unreported (catalog reactivity; catalog IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q3SXY8 (AR13B_HUMAN, ADP-ribosylation factor-like protein 13B).
- Human Protein Atlas. ARL13B tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ARL13B subcellular location (ICC-IF): Mainly localized to the microtubules, primary cilium and primary cilium transition zone. In addition localized to the cytokinetic bridge, basal body, acrosome, equatorial segment, mid piece, principal piece and end piece..
- Human Protein Atlas. ARL13B antibody validation summary (1 antibodies).
- Distinct patterns of primary and motile cilia in Rathke's cleft cysts and craniopharyngioma subtypes. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 2016 — PMC5442446.
- Recombinant Antithrombin Attenuates Acute Respiratory Distress Syndrome in Experimental Endotoxemia. The American journal of pathology 2021 — PMC8278305.
- Arl13b Regulates Breast Cancer Cell Migration and Invasion by Controlling Integrin-Mediated Signaling. Cancers 2019 — PMC6826833.
- Progressive tooth pattern changes in Cilk1-deficient mice depending on Hedgehog signaling. International journal of oral science 2025 — PMC12665794.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:16641997 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ARL13B (ADP-ribosylation factor-like protein 13B). Expected localisation Cytoplasm in tissue; ciliary membrane molecularly (HPA tissue IHC; UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04279-1). Validated IHC antibody: A04279-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular 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 breast cancer tissue; fixative not specified (datasheet A04279-1); 2. Fixation: Image fixative and duration unreported (datasheet A04279-1); 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 A04279-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04279-1); 8. Primary antibody: Rabbit anti-ARL13B, 2-5 μg/ml (datasheet A04279-1); 9. Primary incubation: Overnight at 4 °C (datasheet A04279-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining has medium concordance with RNA (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unknown (UniProt); Regulation: Expressed in the developing brain (UniProt). Evidence: UniProt Q3SXY8, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the microtubules, primary cilium and primary…, Datasheet A04279-1, PMC5442446, PMC6826833. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ARL13B/immunohistochemistry-ihc-guide.