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ARL13B / ADP-ribosylation factor-like protein 13B · IHC design guide
Design Immunohistochemistry for ARL13B
This guide uses tissue IHC evidence to interpret ARL13B’s broad cytoplasmic staining (HPA tissue IHC). It covers the catalog antibody’s 2–5 μg/mL paraffin IHC range (datasheet A04279-1) and the distinct molecular expectation of cilium membrane localisation (UniProt).
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 | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in cells across 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 (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04279-1) | |
| Caveat | Broad cytoplasmic staining may obscure ciliary signal (HPA tissue IHC; UniProt) | |
| Regulation | Expressed in the developing brain (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended ARL13B IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by published ARL13B protocols for tissue sections (PMC5442446; PMC8278305; 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 heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet: EDTA pH 8.0 HIER); a published breast tissue protocol used citrate at pH 6.0 (PMC6826833).
Section 2
What Is the Expected ARL13B Staining Pattern?
ARL13B is associated with the cilium membrane and cytoskeleton and has no annotated transmembrane segment (UniProt Q3SXY8). In paraffin sections, expect cytoplasmic staining across many tissues, with high staining reported in adrenal and gallbladder glandular cells and lung alveolar type I cells (HPA tissue IHC). HPA rates its tissue IHC evidence Approved, with medium consistency between antibody staining and RNA expression (HPA tissue IHC).
| Cytoplasmic stain in adrenal or gallbladder glandular cells, or lung alveolar type I cells. | This matches the cells and high staining levels reported in tissue sections (HPA tissue IHC). A discernible ciliary profile can add support, but its absence in a section does not by itself overturn a matching tissue pattern (UniProt Q3SXY8 localization; general IHC practice). |
| A sharply nuclear pattern dominates while the expected cytoplasmic pattern is absent. | Nuclear localization is outside the supplied ARL13B locations (UniProt Q3SXY8; HPA tissue IHC). Treat this as suspect staining; review the antibody, detection controls and morphology before assigning it to ARL13B (general IHC practice). |
| Strong stain appears in a cell population reported as low, while expected high-staining cells are blank. | For example, respiratory epithelial cells are reported low, whereas lung alveolar type I cells are high (HPA tissue IHC). Check cell identification and controls; cross-reactivity or endogenous detection activity is possible, but this pattern alone cannot distinguish them (general IHC practice). |
| Broad, diffuse colour obscures cell boundaries and the expected cell-to-cell pattern. | The result is difficult to score against the reported cytoplasmic and cell-specific distribution (HPA tissue IHC). Background from detection reagents or insufficient blocking is possible; compare controls before calling the diffuse colour positive (general IHC practice). |
| No stain is visible in an adrenal gland, gallbladder or lung section containing the reported cells. | These are useful positive-reference cells because HPA reports high staining in them (HPA tissue IHC). First confirm that the relevant cells are present, then check the IHC run and antibody conditions; one blank section does not establish biological absence (general IHC practice). |
💡Expected ARL13B appearanceCall positive when identifiable adrenal or gallbladder glandular cells, or lung alveolar type I cells, show clear cytoplasmic staining consistent with HPA's High level; isolated dominant nuclear colour or widespread colour without cell definition is suspect (HPA tissue IHC; general IHC practice).
| Tissue and cell selection | Adrenal and gallbladder glandular cells and lung alveolar type I cells are reported High; endometrial glandular cells and adipocytes are reported Medium (HPA tissue IHC). Compare like cell populations when judging intensity. |
| Compartment and section geometry | ARL13B associates with proximal ciliary membranes and also colocalizes with microtubules (UniProt Q3SXY8). HPA reports cytoplasmic staining in most tissue sections (HPA tissue IHC); a thin cilium may be hard to resolve by chromogenic IHC (general IHC practice). |
| Antibody evidence | The listed antibody HPA048926 has IHC Approved and ICC Supported status (HPA antibodies). Tissue IHC has medium consistency with RNA data (HPA tissue IHC), so use morphology and controls when interpreting an unexpected distribution (general IHC practice). |
| Isoform coverage | Three isoforms are annotated (UniProt Q3SXY8). The supplied record gives no epitope or isoform coverage for the IHC-validated antibody, so a stain cannot be assigned to a particular isoform from these data. |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected high-staining cells are present, but the section is blank. | The result conflicts with HPA's reported High staining in those cell populations (HPA tissue IHC); an IHC run or reagent issue remains possible (general IHC practice). | Check the positive-reference section and detection controls, then review retrieval and antibody dilution under the laboratory's validated IHC conditions (general IHC practice). |
| Colour is mainly nuclear. | This does not match the supplied cytoplasmic tissue pattern or ciliary and cytoskeletal locations (HPA tissue IHC; UniProt Q3SXY8). Background or off-target binding is possible (general IHC practice). | Compare a reagent control and a positive-reference tissue; inspect cell boundaries before scoring the nuclear colour (general IHC practice). |
| Unexpected cells stain more strongly than the reported high-staining cells. | Cell identification, cross-reactivity or endogenous detection activity could account for the mismatch (general IHC practice); HPA's levels apply to named cell populations (HPA tissue IHC). | Recheck morphology and compare appropriate detection controls with the HPA cell-level pattern before attributing the signal to ARL13B (HPA tissue IHC; general IHC practice). |
| Diffuse colour makes cytoplasmic staining hard to distinguish. | Detection background or insufficient blocking may obscure the reported pattern (HPA tissue IHC; general IHC practice). | Inspect a no-primary control and review blocking, washes and chromogen development within the validated IHC workflow (general IHC practice). |
| Ciliary staining is not individually visible in an otherwise plausible section. | Section plane and chromogenic resolution can limit visibility of a small cilium (general IHC practice); HPA describes tissue IHC broadly as cytoplasmic (HPA tissue IHC). | Judge the named cell populations and cytoplasmic distribution first; reserve finer subcellular assessment for an IF/ICC experiment (HPA tissue IHC; general IHC practice). |
| What should IF/ICC show if finer localization is needed? | HPA reports supported localization at microtubules, primary cilia and the primary cilium transition zone in ICC-IF (HPA subcellular). UniProt describes a proximal ciliary membrane compartment that excludes the transition zone (UniProt Q3SXY8). | Use the separate IF/ICC guide for experiment design; interpret transition-zone assignment with the distinction between the HPA imaging summary and UniProt's more specific membrane description in mind (HPA subcellular; UniProt Q3SXY8). |
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 adrenal gland first and assess staining in its glandular cells (HPA: High in adrenal gland glandular cells). HPA detects ARL13B in all 45 scored tissues, so use no-primary and isotype controls for the negative comparison; cells outside the reported glandular compartment should be assessed for background staining, but no cell type on this slide is established as ARL13B-negative (HPA: no negative tissue rows; HPA: High in adrenal gland glandular cells).
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 secondary-only section, an immunoglobulin- and species-matched isotype control for the rabbit primary, and an ARL13B-knockout biological control where available (caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase and check for DAB background in the adrenal section before scoring (caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: The selected paraffin-section caption reports heat retrieval in EDTA at pH 8.0, but gives no fixative; a target-specific fixation window or fixation effect is unreported (caption: EDTA retrieval, pH 8.0; caption: fixative not stated). IF/ICC in serum-starved hTERT-RPE1 cells offers a way to resolve ciliary localization, while the supplied evidence does not establish that frozen sections are easier (HPA: hTERT-RPE1 serum-starved ICC-IF images; HPA: primary cilium localization). In adrenal IHC, distinguish discrete staining from endogenous-peroxidase-related DAB background using the negative controls (caption: HRP/DAB detection; standard IHC 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 staining in paraffin sections by checking retrieval, staining compartment, controls and scoring before interpreting diffuse signal (UniProt Q3SXY8; HPA tissue IHC).
Which retrieval condition should I try first for ARL13B paraffin sections?
Start with heat mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A04279-1). The selected image used this condition before overnight incubation at 4°C with 2 μg/ml antibody, so compare retrieval changes while holding antibody concentration and detection constant (caption A04279-1). If staining is weak, adjust heating duration or cooling within your validated tissue workflow and compare serial sections against a consistently stained control (standard IHC practice). Excessively harsh retrieval can damage morphology and make small ciliary profiles harder to distinguish, so judge signal alongside tissue preservation (standard IHC practice; UniProt Q3SXY8 localisation).
Could fixation explain weak or diffuse ARL13B staining?
The selected paraffin section caption does not report its fixative, so ARL13B sensitivity to fixation is unknown from the supplied IHC evidence (caption A04279-1). Record the fixative, time before fixation, fixation duration and section age for each specimen, then compare matched sections under the same EDTA pH 8.0 retrieval condition (standard IHC practice; datasheet A04279-1). If signal varies across batches, test processing variables with a stable control and assess morphology before changing antibody concentration (standard IHC practice). Do not attribute weak ciliary or cytoplasmic staining to a specific fixation effect without a controlled comparison (standard IHC practice; UniProt Q3SXY8 localisation).
How should I judge ciliary staining alongside broad cytoplasmic staining?
ARL13B associates with the cilium membrane through palmitoylation and is also annotated in the cytoskeleton, so inspect both discrete projections and broader cellular staining (UniProt Q3SXY8). The HPA tissue survey describes cytoplasmic expression in most tissues, while its cell imaging supports primary cilium and microtubule localisation (HPA tissue IHC; HPA subcellular). Use morphology and, where available, serial sections to decide whether a punctum or short projection belongs to a cell rather than overlying debris (standard IHC practice). Interpret an isolated diffuse pattern cautiously when the positive control shows a reproducible, more structured pattern under the same conditions (standard IHC practice; UniProt Q3SXY8 localisation).
Can splice isoforms or epitope access explain inconsistent ARL13B staining?
UniProt lists 3 ARL13B isoforms, but the supplied antibody caption does not identify its binding epitope or establish which isoforms it detects (UniProt Q3SXY8; caption A04279-1). Check the antibody's documented immunogen and map it against the relevant isoform sequences before making an isoform claim (standard IHC practice). ARL13B has no annotated transmembrane segment and associates with the cilium membrane through palmitoylation; those annotations do not establish how retrieval exposes this antibody's epitope (UniProt Q3SXY8). If specimens disagree, compare matched sections using EDTA pH 8.0 retrieval and identical detection before invoking isoform dependent staining (datasheet A04279-1; standard IHC practice).
How can IF clarify an ambiguous ARL13B chromogenic pattern?
Use IF as a separate follow up to resolve candidate ciliary profiles; the selected catalog evidence describes paraffin IHC, not an IF protocol (caption A04279-1). Multiplex ARL13B with a marker of the expected cell type, such as a glandular epithelial marker when assessing glandular cells, and include single label controls (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores, preferably assessing a far red channel when tissue autofluorescence obscures shorter wavelengths (standard IF practice). Because ARL13B lacks a transmembrane segment and associates with ciliary membrane through palmitoylation, assess whether controlled permeabilisation improves access to its intracellular epitope; the antibody's epitope side is unreported (UniProt Q3SXY8; caption A04279-1).
What should I change when DAB obscures ARL13B signal?
The selected paraffin image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase linked secondary followed by DAB (caption A04279-1). First compare a no primary control with the stained section to identify secondary reagent, endogenous enzyme or chromogen background (standard IHC practice). Apply a peroxidase block and optimise wash stringency and DAB development within the laboratory's chromogenic workflow (standard IHC practice). If background remains, titrate the primary around the documented concentration while checking that ciliary or cellular signal is retained in a positive control (caption A04279-1; UniProt Q3SXY8 localisation; standard IHC practice).
How should I score ARL13B staining across heterogeneous sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring, because ARL13B has ciliary and cytoskeletal annotations and HPA reports broad cytoplasmic tissue staining (UniProt Q3SXY8; HPA tissue IHC). For cellular chromogenic signal, report the percentage of positive cells and an H-score based on intensity across the prespecified population (standard IHC practice). For resolvable ciliary profiles, report positive profile density per mm² of evaluable tissue or the fraction of eligible cells with a positive profile (standard IHC practice). Normalise comparisons to the same tissue area or eligible cell count, apply identical thresholds, and exclude damaged or necrotic regions using predefined rules (standard IHC practice).
When is an apparent ARL13B positive result likely to be artefact?
A credible result should follow cellular morphology and be reproducible in the expected compartment across comparable sections (standard IHC practice; UniProt Q3SXY8 localisation). ARL13B is associated with ciliary membrane and cytoskeleton, while HPA also reports cytoplasmic staining in most tissues, so diffuse cytoplasm alone is insufficient to establish a cilium specific pattern (UniProt Q3SXY8; HPA tissue IHC). Compare candidate signal with the expected cell population and examine section edges, folds and necrotic areas for staining unrelated to intact cells (HPA tissue IHC; standard IHC practice). A no primary control helps identify endogenous peroxidase or detection artefact when DAB deposits appear in unexpected compartments (standard IHC practice).
Boster reagents
Best ARL13B / ADP-ribosylation factor-like protein 13B IHC Antibodies
Anti-ARL13B A04279-1 has IHC images from human paraffin-embedded breast, colorectal, gastric, and testicular tumor sections (IHC image captions) and an IF image from U20S cells (IF image caption).
Real IHC data
Anti-ARL13B Antibody ®
Cat # A04279-1
A04279-1 will render with its human breast cancer paraffin-section IHC image; the catalog also shows colorectal, gastric, and testicular cancer paraffin-section IHC images (IHC image captions). Its IF image shows U20S cells (IF image caption), and its listed reactivity is human, mouse, and rat (catalog: reactivity).
Which to pick: Choose A04279-1 for paraffin-section tissue IHC: its human breast cancer image used EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (IHC image caption); the fixative is unreported (IHC image caption). For IF/ICC, A04279-1 is listed for both applications and has an IF image from U20S cells (catalog: applications; IF image caption). For mouse or rat work, A04279-1 lists those species as reactive, while its supplied IHC images document human tissue only (catalog: reactivity; IHC image 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 Cytoplasmic staining in most tissues (HPA tissue IHC). 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 (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04279-1); Caveat: Broad cytoplasmic staining may obscure ciliary signal (HPA tissue IHC; UniProt); Isoform / epitope: 3 isoforms; epitope coverage is unspecified (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, PMC8278305. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ARL13B/immunohistochemistry-ihc-guide.