TRANK1 / TPR and ankyrin repeat-containing protein 1 · IHC design guide

Design Immunohistochemistry for TRANK1

Plan chromogenic TRANK1 IHC in paraffin sections using the reported ciliary and cytoplasmic tissue pattern (HPA tissue IHC). The guide covers the 2–5 μg/ml antibody range (datasheet A09518) and interpretation of staining with low antibody–RNA consistency (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for TRANK1 (IHC for TRANK1): expected localisation Cilia and cytoplasm in tissue staining (HPA tissue IHC), antibody A09518, validated IHC image, and IHC protocol steps
Printable TRANK1 IHC protocol sheet — expected localisation Cilia and cytoplasm in tissue staining (HPA tissue IHC), antibody A09518, controls and protocol steps. Open the full TRANK1 IHC guide →

TRANK1 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 Cilia and cytoplasm in tissue staining (HPA tissue IHC)
Staining pattern Strong ciliary staining in ciliated cells; cytoplasm in several tissues (HPA tissue IHC)
Antigen retrieval EDTA pH 8.0 HIER, heat-mediated (datasheet A09518)
Positive control ⓘ Bronchus+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); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A09518)
Caveat Antibody staining and RNA expression show low consistency (HPA tissue IHC)
Regulation No expression regulator annotated (UniProt)
Isoform / epitope 0 isoforms annotated; epitope location unspecified (UniProt)
Section 1

Recommended TRANK1 IHC & IF Protocols

The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet A09518). The published rat prefrontal cortex protocol uses citrate pH 6.0 (PMC5355521).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human liver cancer tissue; fixative not specified (datasheet A09518)
FixationImage fixative and duration unreported (datasheet A09518); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: EDTA pH 8.0 (datasheet A09518); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet A09518)
Primary antibodyRabbit anti-TRANK1, 2-5 μg/ml (datasheet A09518)
Primary incubationOvernight at 4 °C (datasheet A09518)
DetectionHRP-conjugated secondary, DAB chromogen (datasheet A09518)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultTRANK1-positive staining in ciliated cells (cilia axoneme) of bronchus (HPA tissue IHC: High). HPA tissue profile: Distinct expression in cilia in fallopian tube, respiratory epithelia and efferent ducts. Cytoplasmic expression in several tissues. No signal in the no-primary control.
💡Decision noteStart with EDTA pH 8.0 for the catalog antibody (datasheet A09518); use citrate pH 6.0 when reproducing the published rat protocol (PMC5355521).
Section 2

What Is the Expected TRANK1 Staining Pattern?

In paraffin-section IHC, expect strong staining along the cilia axoneme of ciliated cells in bronchus, nasopharynx and fallopian tube, with high staining also reported in efferent-duct ciliated cells (HPA: tissue IHC). Several tissues show cytoplasmic expression (HPA: tissue IHC). TRANK1 has no annotated transmembrane segment (UniProt O15050 topology). HPA rates its tissue staining Approved but reports low agreement with RNA data and pending external verification (HPA: reliability).

What am I looking at on my slide?
A distinct apical ciliary signal appears in bronchus, nasopharynx or fallopian tube, with little stain outside the ciliated cells.This matches the strongest, most specific tissue pattern reported for TRANK1: High staining in ciliated-cell cilia axonemes (HPA: tissue IHC). Judge both the cell type and the fine ciliary location; overall brown intensity alone is less informative. The HPA tissue profile remains pending external verification (HPA: reliability).
The intended ciliary signal is replaced by nuclear-only or uniformly diffuse staining in ciliated epithelium.That does not reproduce the cilia-axoneme pattern reported for these tissues (HPA: tissue IHC). Review morphology, counterstain and staining controls before calling it specific (general IHC practice). Nuclear staining alone is not proof of an artefact: HPA separately reports approved nucleoplasmic localisation in ICC-IF (HPA: subcellular ICC-IF).
Strong stain appears mainly in an HPA-listed negative cell population while nearby ciliated cells lack the expected pattern.Treat this as a specificity warning, not an automatic TRANK1-positive call. HPA reports Not detected in alveolar cells and adipocytes, for example (HPA: tissue IHC). Cross-reactivity or endogenous detection activity are possible technical explanations; compare omission and detection controls before assigning a cause (general IHC practice).
A broad haze covers tissue, stroma or blank regions and obscures cell boundaries.A slide-wide haze cannot establish the cell-restricted ciliary pattern (HPA: tissue IHC). Uneven washing, insufficient blocking or detection background can cause diffuse chromogenic signal (general IHC practice). Compare a primary-antibody omission control and inspect whether fine cilia remain distinguishable.
No ciliary signal is visible in a well-preserved bronchus, nasopharynx or fallopian-tube section.This conflicts with HPA's High cilia-axoneme staining in those tissues (HPA: tissue IHC). First confirm that ciliated cells are present in the section; then assess antibody, retrieval and detection performance with appropriate run controls (general IHC practice). An absent signal in one run does not establish biological absence.
💡Expected TRANK1 appearanceCall an IHC result convincing when High signal follows the cilia axoneme of identifiable ciliated cells in bronchus, nasopharynx or fallopian tube (HPA: tissue IHC); isolated diffuse, stromal or wrong-cell staining without that pattern is a specificity warning (general IHC interpretation).
How each factor affects the staining
Tissue and cell selectionUse ciliated epithelium in bronchus, nasopharynx or fallopian tube to assess the defining axonemal pattern (HPA: High in ciliated-cell cilia axoneme). Efferent-duct ciliated cells are another High-staining population, while adrenal, breast and endometrial glandular cells are Medium (HPA: tissue IHC). Compare named cell populations, since an HPA tissue page does not imply every cell in that tissue stains.
Validation and strength of inferenceHPA lists one antibody, HPA035508, with IHC Approved status (HPA: antibodies). Its tissue assessment reports low consistency with RNA expression and pending external verification (HPA: reliability). Treat the published pattern as a working expectation and weigh morphology and controls before making a biological claim; Approved does not remove the reported caveat.
TRANK1 topology and processingUniProt annotates no transmembrane segment, signal peptide, propeptide, glycosylation site or modified residue, and lists a single chain spanning residues 1–2925 (UniProt O15050 record). These annotations offer no basis to predict a membrane-restricted IHC pattern or a processing-dependent staining difference. No target-specific fixation effect follows from them (UniProt O15050 record).
IF/ICC Q&A: should nuclear fluorescence change the IHC call?HPA reports nucleoplasm as the main approved ICC-IF location and cytosol as an additional approved location (HPA: subcellular ICC-IF). Interpret that observation in its own assay; the tissue IHC comparison remains the ciliary and cytoplasmic pattern reported by HPA (HPA: tissue IHC). UniProt provides no subcellular annotation to resolve the difference (UniProt O15050 record).
Why is my staining missing, weak or wrong?
SituationLikely causeNext action
No stain appears in the intended positive section.The section may lack intact ciliated cells, or the IHC detection run may have failed (general IHC practice). HPA reports High axonemal staining in bronchus, nasopharynx and fallopian tube (HPA: tissue IHC).Locate preserved cilia on the counterstained section, then check run controls, antibody application, retrieval and chromogen steps (general IHC practice). Record the missing control pattern before interpreting the specimen as negative.
Brown product fills epithelium but individual cilia cannot be resolved.Excess background or heavy development can obscure the fine axonemal pattern (general IHC practice). The expected location is the cilia axoneme in several High-staining tissues (HPA: tissue IHC).Examine a lighter-developed or better-separated area; review primary-antibody concentration, wash steps and development time with run controls (general IHC practice). Score localisation only where morphology remains readable.
Negative-control areas or blank regions also develop colour.Endogenous detection activity or reagent background may contribute signal (general IHC practice). A control-area signal does not support the HPA cell-restricted pattern (HPA: tissue IHC).Compare primary-antibody omission and detection controls; review blocking and detection chemistry for the system used (general IHC practice). Do not count colour shared with those controls as specific TRANK1 staining.
A listed negative population stains strongly.Cross-reactivity, background or a biological difference may be involved; the slide alone cannot distinguish them. HPA lists alveolar cells, adipocytes and several other named populations as Not detected (HPA: tissue IHC).Verify the cell identity, compare matched controls and assess whether the stain also occurs in a cilia-positive reference (general IHC practice). Report disagreement with the HPA reference instead of assigning a cause from intensity alone.
Staining is predominantly nuclear in tissue IHC.The observed compartment differs from HPA's defining axonemal tissue pattern (HPA: tissue IHC), although approved nucleoplasmic staining is reported for ICC-IF (HPA: subcellular ICC-IF).Keep the IHC and ICC-IF observations distinct. Check section morphology, counterstain and assay controls before deciding whether nuclear IHC signal is interpretable (general IHC practice).
Two acceptable sections give different staining intensities.Cell composition and staining-run variation can alter the appearance of a section (general IHC practice). HPA reports High, Medium, Low and Not detected levels in different named cell populations (HPA: tissue IHC).Compare the same cell type and compartment, then review controls from each run (general IHC practice). Document tissue and cell identity alongside intensity; avoid treating unlike cell populations as equivalent replicates.

Sample controls for TRANK1 IHC & IF

🧪Run bronchus first and look for staining in the cilia axonemes of ciliated cells (HPA: High in bronchus ciliated cells). Use esophageal squamous epithelial cells as the tissue negative (HPA: Not detected); within the bronchus slide, nonciliated cells should lack specific axonemal staining and provide a local background comparison (HPA: signal assigned to ciliated-cell cilia axonemes).
Positive control tissue: Bronchus (Ciliated cells (cilia axoneme), HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TRANK1 in A-431, U-251MG, U2OS, ASC52telo, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a concentration-matched rabbit isotype control, and a TRANK1 knockout sample as a biological negative (caption: rabbit primary antibody; standard IHC control practice). Quench endogenous peroxidase and inspect the bronchus slide for background before interpreting DAB staining (caption: peroxidase detection with DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A09518 tissue-IHC caption does not state a fixative (selected-SKU caption: fixative not stated). The demonstrated paraffin-section procedure used heat retrieval in EDTA at pH 8.0, so begin there; whether retrieval is essential, or frozen sections or IF are easier, is unreported (selected-SKU caption: EDTA heat retrieval). Interpret staining at the cilia axoneme against adjacent background, where the small structure makes nonspecific edge staining a practical concern (HPA: High in bronchus cilia axoneme; standard IHC interpretation practice).

HPA tissue IHC evidence for TRANK1

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. 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

TissueCell typeLevelEvidenceSource
Bronchus Ciliated cells (cilia axoneme) High Protein (IHC) HPA →
Choroid plexus Ependymal cells High Protein (IHC) HPA →
Efferent ducts Ciliated cells High Protein (IHC) HPA →
Fallopian tube Ciliated cells (cilia axoneme) High Protein (IHC) HPA →
Nasopharynx Ciliated cells (cilia axoneme) High Protein (IHC) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Adipose tissue Adipocytes Not detected Protein (IHC) HPA →
Appendix Glandular cells Not detected Protein (IHC) HPA →
Cerebral cortex Endothelial cells Not detected Protein (IHC) HPA →
Esophagus Squamous epithelial cells Not detected Protein (IHC) HPA →
Heart muscle Cardiomyocytes Not detected Protein (IHC) HPA →
Section 3

Advanced TRANK1 IHC Tips

Use matched controls and compartment-aware scoring to troubleshoot TRANK1 staining in paraffin sections; interpret tissue patterns alongside the reported limits of antibody evidence.

How should I adjust retrieval when TRANK1 staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A09518). The selected tissue image used this retrieval before overnight incubation with 2 μg/ml primary antibody at 4°C (caption A09518). If staining remains weak, check deparaffinisation, retrieval heating and section integrity, then compare a small range of retrieval times on adjacent sections (standard IHC practice). Keep antibody concentration and DAB development constant during that comparison so a stronger signal can be attributed to retrieval (standard IHC practice). Include ciliated epithelium as a reference, while recognising the reported ciliary staining has limited cross-method agreement (HPA: high in ciliated cells; HPA: low staining–RNA consistency).
Could fixation explain variable TRANK1 staining between paraffin blocks?
Target-specific fixation sensitivity is unknown because the selected paraffin-section caption does not state its fixative (caption A09518). Record the actual fixative, fixation duration, tissue thickness and processing history for each block before attributing a staining difference to TRANK1 biology (standard IHC practice). Compare adjacent sections from similarly processed blocks using the same EDTA pH 8.0 retrieval and primary concentration of 2 μg/ml (datasheet A09518; caption A09518). If a block stains poorly, inspect morphology and include a process control to assess whether antigen preservation or processing failed more broadly (standard IHC practice). Neither the observed tissue pattern nor the protein’s lack of a transmembrane segment establishes a fixation effect (HPA: tissue IHC; UniProt O15050 topology).
Which compartment should count as a credible TRANK1 signal?
Score the compartment explicitly: tissue IHC reports distinct ciliary staining in several ciliated epithelia and cytoplasmic expression in several tissues (HPA: tissue IHC profile). Cell-based IF instead reports mainly nucleoplasmic signal with additional cytosolic signal, so those observations should guide review without being imposed as identical tissue-IHC patterns (HPA: subcellular). UniProt does not annotate a subcellular location or a transmembrane segment for TRANK1 (UniProt O15050). Compare candidate staining with cell morphology and an adjacent negative-control section, especially where thin cilia could be mistaken for luminal precipitate (standard IHC practice). Record nuclear, cytoplasmic and ciliary signal separately rather than combining them into one positive call (standard IHC practice).
Could an unrecognised isoform or masked epitope explain discordant staining?
The supplied UniProt record lists 0 annotated isoforms and gives no epitope map; this does not establish that every biological variant is absent (UniProt O15050). It also lists no signal peptide, propeptide or annotated glycosylation sites, so do not assign an unexplained staining difference to a specific cleavage or glycan without direct evidence (UniProt O15050). For the catalog antibody, first reproduce EDTA pH 8.0 retrieval and 2 μg/ml primary incubation used in its paraffin-section image (caption A09518). If discordance persists, compare independently recognised epitopes on serial sections, with matched controls and recorded compartment patterns (standard IHC practice). Treat agreement between antibodies as supporting evidence, rather than proof of molecular identity (standard IHC practice).
How can IF help investigate ambiguous chromogenic TRANK1 staining?
On a separately validated IF/ICC workflow, multiplex TRANK1 with a marker for the expected ciliated cell population when testing the tissue-IHC ciliary pattern (HPA: high in ciliated cells; standard IF practice). Select fluorophores in channels with low tissue autofluorescence, and inspect unstained and single-label controls before interpreting overlap (standard IF practice). For intracellular nucleoplasmic or cytosolic epitopes, assess permeabilisation after fixation; the supplied record provides no mapped epitope or membrane-sided orientation to prescribe a target-specific treatment (HPA: subcellular; UniProt O15050). Compare IF with paraffin-section IHC as complementary localisation evidence, since the selected antibody image documents paraffin-section DAB staining, while cell-based IF reports nucleoplasmic and cytosolic locations (caption A09518; HPA: subcellular).
What should I change when DAB appears throughout the section?
First inspect a no-primary section to separate secondary-reagent or endogenous-peroxidase signal from staining that depends on the primary antibody (standard IHC practice). The selected image used a peroxidase-conjugated secondary and DAB, making a peroxidase block and controlled chromogen development relevant workflow checks (caption A09518; standard IHC practice). Confirm the 10% goat-serum block and 2 μg/ml primary concentration before titrating either variable on adjacent sections (caption A09518). Review wash stringency, tissue drying and precipitate, especially at section edges and luminal surfaces where deposits can mimic ciliary signal (standard IHC practice). Judge residual staining against cell morphology and a matched control, not against overall section darkness alone (standard IHC practice).
How should I quantify TRANK1 staining across heterogeneous tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because reported tissue staining includes ciliary and cytoplasmic patterns (HPA: tissue IHC profile). For broad cellular staining, report percentage of positive cells and an intensity-weighted H-score; for thin ciliary profiles, report positive ciliated cells or stained ciliary length per assessed area (standard IHC practice). Normalise counts to eligible cells or evaluable tissue area in mm², and exclude folds, necrosis and detached edges using a prespecified rule (standard IHC practice). Keep retrieval, 2 μg/ml primary concentration, imaging and DAB development consistent across the comparison (caption A09518; standard IHC practice). Report nuclear and cytoplasmic scores separately if those compartments are under study (HPA: subcellular; standard IHC practice).
When is an apparent TRANK1-positive cell likely an artefact?
A credible tissue call should match identifiable cells and a reproducible compartment pattern, such as the reported ciliary staining in ciliated epithelia (HPA: tissue IHC profile; standard IHC practice). Diffuse edge staining, necrotic deposits and signal persisting without primary antibody favour processing or detection artefact (standard IHC practice). Unexpected strong staining in a population reported as undetected, or a compartment inconsistent with the cell-based nucleoplasmic and cytosolic observations, warrants review rather than automatic rejection (HPA: tissue IHC; HPA: subcellular). Check an adjacent no-primary section for endogenous-peroxidase or secondary-reagent signal and repeat questionable regions with controlled DAB development (standard IHC practice). Treat a positive image as provisional because the tissue-IHC profile is approved yet reports low staining–RNA consistency and pending external verification (HPA: reliability description).
Boster reagents

Best TRANK1 / TPR and ankyrin repeat-containing protein 1 IHC Antibodies

Anti-TRANK1 antibody A09518 has IHC images from paraffin-embedded human liver cancer and mouse and rat brain tissue, plus IF images from those tissues and PC-3 cells (A09518 image captions).

Real IHC data IHC analysis of TRANK1 using anti-TRANK1 antibody (A09518). TRANK1 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-TRANK1 Antibody (A09518) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Anti-TRANK1 Antibody ®
Cat # A09518

A09518 is listed for IHC and IF/ICC in human, mouse and rat (A09518 catalog applications and reactivity). Its IHC images show paraffin-embedded human liver cancer and mouse and rat brain tissue; its IF images also show PC-3 cells (A09518 image captions).

Which to pick: Choose A09518 for paraffin-section IHC: its captions document EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (A09518 IHC image captions). The same SKU supports IF/ICC at 5 μg/ml, with IF images from PC-3 cells and paraffin-embedded tissue (A09518 catalog applications and IF image captions). For cross-species IHC, A09518 lists human, mouse and rat reactivity and shows IHC in all three; the IHC captions do not report the fixative (A09518 catalog reactivity and IHC image captions).

Each figure is that product's own IHC / IF validation image from its datasheet.