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TTBK2 / Tau-tubulin kinase 2 · IHC design guide
Design Immunohistochemistry for TTBK2
Plan chromogenic IHC on paraffin sections using bronchus or fallopian tube ciliated cells as high-staining controls (HPA tissue IHC). Start the catalog antibody A05821 at 5 μg/mL (datasheet: A05821) and assess cytoplasmic and ciliary-tip staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TTBK2 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 and ciliary tips in tissue (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining; ciliary tips in ciliated cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| 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 paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining has low consistency with RNA data (HPA tissue IHC) | |
| Regulation | Testis-enhanced RNA (HPA tissue RNA) | |
| Isoform / epitope | 3 isoforms; no membrane segment; epitope coverage unknown (UniProt) |
Section 1
Recommended TTBK2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with published TTBK2 tissue IHC protocols (PMC4256087; PMC5802059).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A05821); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-TTBK2, 5 μg/mL (datasheet A05821) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TTBK2-positive staining in ciliated cells (tip of cilia) of bronchus (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues including cilia in fallopian tube and sperm in testis. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA HIER at pH 9.0, 95–98 °C for 20 min (page retrieval); compare citrate autoclaving for human FFPE brain (PMC5802059).
Section 2
What Is the Expected TTBK2 Staining Pattern?
TTBK2 is associated with the ciliary transition zone and basal body, with additional cytosolic and lower-level nuclear localization (UniProt Q6IQ55). In tissue IHC, expect strong staining at cilia tips in ciliated bronchial, fallopian-tube and nasopharyngeal cells, alongside cytoplasmic staining in several tissues (HPA: tissue IHC). Interpret the pattern cautiously: HPA rates the staining Approved but reports low consistency with RNA expression (HPA: tissue IHC). TTBK2 has no transmembrane segment (UniProt Q6IQ55 topology).
| Strong, localized staining at cilia tips in bronchus, fallopian tube or nasopharynx. | This matches the reported high signal in ciliated cells of all three tissues (HPA: tissue IHC). Evaluate the stained structure and cell identity together; intensity alone does not establish specificity (general IHC practice). |
| Predominantly sharp cell-surface rims or extracellular deposits, without the expected ciliary or cytoplasmic pattern. | That distribution conflicts with TTBK2's reported intracellular locations and lack of a transmembrane segment (UniProt Q6IQ55). Treat it as suspect and assess controls for staining artefact (general IHC practice). Weak nuclear staining alone is less decisive because lower-level nuclear localization is reported (UniProt Q6IQ55). |
| Strong staining in a cell population reported as undetected, such as adipocytes or glomerular cells. | Those populations are listed as Not detected (HPA: tissue IHC). Check morphology and controls for cross-reactivity or endogenous detection activity (general IHC practice). An individual discrepant specimen needs cautious interpretation because HPA reports low staining–RNA consistency (HPA: tissue IHC). |
| Uniform haze across tissue, empty spaces and the negative control. | A pattern that does not follow cells or cilia is consistent with nonspecific background (general IHC practice). Review blocking, washes, detection reagents and counterstain before assigning a TTBK2 score (general IHC practice). |
| No staining in ciliated bronchial, fallopian-tube or nasopharyngeal epithelium. | Absence conflicts with the reported high cilia-tip staining in these cells (HPA: tissue IHC), but it does not by itself prove target absence. Confirm that cilia are present in the section, then inspect the assay controls and workflow (general IHC practice). |
💡Expected TTBK2 appearanceCall a section positive when ciliated bronchial, fallopian-tube or nasopharyngeal cells show high cilia-tip staining (HPA: tissue IHC); a broad extracellular deposit or isolated membrane rim is discordant with the reported intracellular localization (UniProt Q6IQ55).
| Anatomic and cellular context | HPA reports high cilia-tip staining in three ciliated epithelia, but medium staining in breast myoepithelial cells, epididymal and gallbladder glandular cells, and lung macrophages (HPA: tissue IHC). Match the expected intensity and compartment to the cell being scored (general IHC practice). |
| Antibody evidence | The listed antibody HPA018113 is IHC Approved; its ICC status is Supported (HPA: antibodies). HPA also reports low consistency between tissue staining and RNA expression, so an Approved label alone should not settle an ambiguous cellular pattern (HPA: tissue IHC). |
| Isoforms and epitope uncertainty | Three isoforms are listed (UniProt Q6IQ55). The supplied sources do not locate the antibody epitope or establish recognition of each isoform (HPA: antibodies; UniProt Q6IQ55). Avoid using a tissue stain to infer which isoform is present. |
| Topology and processing | TTBK2 has no transmembrane segment, signal peptide or propeptide; the annotated chain spans residues 1–1244 (UniProt Q6IQ55). These annotations support an intracellular interpretation but do not establish a fixation or antigen-retrieval effect. |
| IF/ICC cross-check — what pattern is reported? | HPA reports mainly cytosol and microtubules, with additional primary cilium, transition-zone and basal-body localization (HPA: subcellular ICC-IF). This is a localization cross-check; the tissue IHC result must still be judged in its own cellular context (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected cilia-tip signal is absent. | The section may not show intact ciliated cells, or the IHC workflow may have failed (general IHC practice). | Inspect cilia and morphology; review positive and negative controls, primary-antibody application, antigen retrieval and detection steps (general IHC practice). Use the reported high-staining ciliated tissues as the pattern reference (HPA: tissue IHC). |
| Brown signal blankets the section. | Background from insufficient blocking or washing, or from endogenous detection activity, may obscure cellular staining (general IHC practice). | Compare a control without primary antibody; review blocking, washes and the detection system, then score only signal that resolves to identifiable cells or cilia (general IHC practice). |
| Nuclear staining dominates. | Lower-level nuclear TTBK2 is reported, while cytosolic and ciliary-associated sites are also reported (UniProt Q6IQ55). A dominant nuclear-only pattern is therefore discordant with that distribution. | Recheck compartment assignment against morphology and controls; avoid calling nuclear intensity alone a convincing positive result (general IHC practice). |
| Strong signal appears in an HPA Not detected cell population. | Cross-reactivity or endogenous detection activity is possible (general IHC practice); HPA lists adipocytes and glomerular cells as Not detected (HPA: tissue IHC). | Confirm cell identity, compare controls and review the staining distribution before reporting a discrepancy (general IHC practice). Note HPA's low staining–RNA consistency when interpreting it (HPA: tissue IHC). |
| Only diffuse cytoplasmic staining is visible in ciliated epithelium. | Cytoplasmic expression is reported, but the high-intensity tissue landmark is the cilia tip in the named epithelia (HPA: tissue IHC). | Check whether cilia and their tips are visible in the section before judging the missing focal pattern; record diffuse and focal staining separately (general IHC practice). |
| Medium staining is mistaken for a failed positive control. | Several reported cell populations have medium rather than high staining, including lung macrophages and breast myoepithelial cells (HPA: tissue IHC). | Judge intensity against the specific cell population; use the reported high-staining ciliated epithelia when assessing a missing strong-signal pattern (HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing TTBK2 staining
A05821 · IHCImmunohistochemistry of TTBK2 in human placenta tissue with TTBK2 antibody at 5 μg/mL.
A05821-1 · IHCImmunohistochemistry of paraffin-embedded human gastric using TTBK2 antibody at dilution of 1:200 (40x lens).
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000128881-TTBK2/tissue/cerebral+cortex#imid_4879904
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000128881-TTBK2/tissue/colon#imid_4879822
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000128881-TTBK2/tissue/liver#imid_4879823
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000128881-TTBK2/tissue/kidney#imid_4879824
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018113; green = TTBK2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018113; green = TTBK2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TTBK2 IHC & IF
🧪Run bronchus first: ciliated cells should stain at the ciliary tips (HPA: bronchus, ciliated cells, High). Use adipose tissue as the negative, where adipocytes are not detected (HPA: adipose tissue, adipocytes, Not detected); on the bronchus slide, adjacent regions without ciliary tips should lack the tip-localized pattern (HPA: bronchus, ciliated cells, tip of cilia).
Positive control tissue: Bronchus (Ciliated cells (tip of cilia), HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TTBK2 in A-431, U-251MG, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), Sperm, with annotated localisation: Microtubules (approved), Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and isotype controls matched to the primary antibody’s host species and immunoglobulin class, plus TTBK2 knockout tissue or a cognate peptide-block control where available (standard IHC practice). Check the bronchus no-primary slide for endogenous peroxidase background before interpreting chromogenic staining (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and antigen-retrieval dependency are unreported in the supplied evidence; the selected placenta IHC caption gives 5 μg/mL but does not state a fixative (A05821 caption: 5 μg/mL; fixative not stated). IF may make ciliary localization easier to resolve, but the supplied evidence does not establish whether IF or frozen sections are easier than paraffin IHC (HPA: bronchus ciliated-cell tips, High; HPA subcellular: primary cilium). In bronchus, distinguish ciliary-tip staining from background in mucus and adjacent epithelium (HPA: bronchus ciliated-cell tips, High; standard IHC practice).
HPA tissue IHC evidence for TTBK2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Bronchus | Ciliated cells (tip of cilia) | High | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (tip of cilia) | High | Protein (IHC) | HPA → |
| Nasopharynx | Ciliated cells (tip of cilia) | High | Protein (IHC) | HPA → |
| Appendix | Lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Breast | Myoepithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TTBK2 IHC Tips
Troubleshoot TTBK2 staining in paraffin sections by checking retrieval, compartment, controls and scoring before interpreting chromogenic signal.
Which retrieval conditions should I start with for TTBK2 paraffin sections?
Start with Tris-EDTA at pH 9.0 for heat-induced retrieval at 95–98 °C for 20 minutes (page retrieval setting). Allow sections to cool consistently, then compare a known positive ciliated region with a matched negative control in the same run (standard IHC practice; HPA: high staining at cilia tips in bronchus and fallopian tube). If staining is weak, optimise retrieval time on adjacent sections before considering another buffer, because retrieval changes can alter both signal and tissue integrity (standard IHC practice). Check whether signal resolves at cilia tips rather than judging intensity from diffuse cytoplasm alone (HPA: cilia-tip staining; UniProt Q6IQ55: transition-zone localisation).
Could fixation explain weak or uneven TTBK2 staining?
TTBK2-specific sensitivity to fixative type or fixation duration is unknown from the supplied evidence (selected A05821 tissue-IHC caption: fixative not stated). Record the fixative and processing history for each paraffin block, and compare sections processed together when troubleshooting uneven staining (standard IHC practice). Inspect morphology and staining across the section, since uneven processing can complicate interpretation of a focal ciliary signal (standard IHC practice; HPA: cilia-tip staining). Keep retrieval, antibody concentration and detection conditions constant while comparing blocks, so a processing difference is easier to identify (standard IHC practice). Do not assign a TTBK2-specific fixation effect without a direct comparison (supplied evidence: no target-specific fixation study).
Where should convincing TTBK2 staining appear in tissue?
Prioritise discrete staining at cilia tips in suitable ciliated tissue, where high TTBK2 signal is reported in bronchus, fallopian tube and nasopharynx (HPA: tissue IHC). A basal-body or transition-zone pattern is biologically plausible, while broader cytosolic and microtubule-associated signal can also occur (UniProt Q6IQ55: transition-zone localisation; HPA: subcellular localisation). Examine the relevant cell and structure at appropriate magnification before calling a tiny punctum positive (standard IHC practice). Nuclear staining alone deserves caution because nuclear TTBK2 is described at a lower level than cytosolic signal (UniProt Q6IQ55: subcellular localisation). Compare compartment patterns with a no-primary control and neighbouring cells on the same section (standard IHC practice).
How can an unknown antibody epitope affect my TTBK2 result?
TTBK2 has 3 annotated isoforms, but the supplied caption does not identify the A05821 epitope or establish which isoforms it detects (UniProt Q6IQ55: isoforms 1–3; selected A05821 tissue-IHC caption). Check the antibody’s documented immunogen or epitope against the isoforms relevant to the study before treating absent staining as absent protein (standard IHC practice). The kinase domain spans residues 21–284, and annotated phosphoserines include residues 445, 786 and 1103 (UniProt Q6IQ55: domains and modified residues). Those annotations alone cannot establish epitope accessibility after processing or prove phosphorylation-specific recognition (standard IHC practice). Confirm consequential differences with an independently validated reagent or orthogonal measurement (standard IHC practice).
How should I check a TTBK2 pattern by multiplex immunofluorescence?
For a separate IF/ICC experiment, pair TTBK2 with a marker identifying the expected ciliated cell or ciliary structure, and inspect whether the signals occupy the expected compartments (HPA: cilia-tip tissue staining; UniProt Q6IQ55: transition-zone localisation; standard IF practice). Choose spectrally separated fluorophores and place a bright reporter in a channel with low measured tissue autofluorescence (standard IF practice). TTBK2 has no annotated transmembrane segment and is reported at intracellular ciliary, basal-body and cytosolic sites, so optimise permeabilisation to expose the antibody’s documented intracellular epitope (UniProt Q6IQ55: topology and localisation; standard IF practice). Include single-stain and no-primary controls when assessing bleed-through and background (standard IF practice).
What should I check when TTBK2 DAB staining looks diffuse?
First compare the DAB pattern with a no-primary control and inspect whether deposits follow tissue edges, folds or damaged areas (standard IHC practice). Block endogenous peroxidase before chromogenic detection, and optimise protein blocking and washes if background persists (standard IHC practice). TTBK2 can appear in cytosol and along microtubules, so diffuse cytoplasmic colour cannot be rejected solely because it is not a ciliary punctum (HPA: subcellular localisation). Assess whether distinct cilia-tip staining survives a cautious reduction in antibody concentration or detection strength (HPA: high cilia-tip staining in several tissues; standard IHC practice). The A05821 placenta image used 5 μg/mL, which documents that image’s condition rather than a universal working concentration (selected A05821 tissue-IHC caption).
How should I score focal TTBK2 staining without losing its ciliary pattern? ⚠ ANSWER MARKED FOR VERIFICATION
Define the region and cell population before scoring, then record the percentage of positive cells and staining intensity for an H-score when the cellular pattern supports that measure (standard IHC practice). For focal ciliary puncta, also consider positive puncta per mm² or per counted ciliated cell, normalised to evaluable tissue area or the number of ciliated cells (HPA: cilia-tip staining; standard IHC practice). Score ciliary and diffuse cytoplasmic compartments separately because both are reported TTBK2 locations (HPA: tissue IHC and subcellular localisation). Apply one threshold across matched sections, exclude damaged areas using preset criteria, and report the denominator alongside each score (standard IHC practice).
How do I distinguish true TTBK2 staining from a tissue artefact?
A credible focal result should match an expected cell and compartment, such as cilia-tip signal in ciliated bronchial, fallopian-tube or nasopharyngeal cells (HPA: high cilia-tip tissue staining). Cytosolic staining can be compatible with TTBK2, but isolated strong nuclear colour requires caution because nuclear presence is reported at a lower level (HPA: cytosolic localisation; UniProt Q6IQ55: lower-level nuclear localisation). Compare candidate signal with no-primary and endogenous-peroxidase controls, especially where DAB deposits follow edges or damaged tissue (standard IHC practice). Exclude necrotic and poorly preserved regions from interpretation, and seek reproducible staining in an independently processed section when the pattern is ambiguous (standard IHC practice).
Boster reagents
Best TTBK2 / Tau-tubulin kinase 2 IHC Antibodies
Two rabbit polyclonal anti-TTBK2 antibodies have real IHC images from human tissue: placenta for A05821 and paraffin-embedded gastric tissue for A05821-1 (catalog; respective image captions).
Real IHC data
Anti-Tau-tubulin kinase 2 TTBK2 Antibody
Cat # A05821
Real IHC data
Anti-Tau-tubulin kinase 2 TTBK2 Antibody
Cat # A05821-1
A05821 lists human IHC-P and shows human placenta staining at 5 μg/mL (catalog; A05821 image caption). A05821-1 lists human and rat IHC reactivity and shows paraffin-embedded human gastric staining at 1:200 (catalog; A05821-1 image caption).
Which to pick: For tissue IHC, choose rabbit polyclonal A05821 for the human placenta example or rabbit polyclonal A05821-1 for the paraffin-embedded human gastric example; neither caption reports a fixative (catalog; respective image captions). For IF/ICC, neither SKU lists that application or provides an IF image (catalog applications and image alts). For rat tissue, A05821-1 is the only listed rat-reactive option, though its IHC image shows human tissue (catalog reactivity; A05821-1 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q6IQ55 (TTBK2_HUMAN, Tau-tubulin kinase 2).
- Human Protein Atlas. TTBK2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. TTBK2 subcellular location (ICC-IF): Mainly localized to the cytosol and microtubules. In addition localized to the primary cilium, primary cilium transition zone and basal body..
- Human Protein Atlas. TTBK2 antibody validation summary (1 antibodies).
- The tau tubulin kinases TTBK1/2 promote accumulation of pathological TDP-43. PLoS genetics 2014 — PMC4256087.
- Pathological phosphorylation of tau and TDP-43 by TTBK1 and TTBK2 drives neurodegeneration. Molecular neurodegeneration 2018 — PMC5802059.
- TTBK2 circular RNA promotes glioma malignancy by regulating miR-217/HNF1β/Derlin-1 pathway. Journal of hematology & oncology 2017 — PMC5319142.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16572171 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TTBK2 (Tau-tubulin kinase 2). Expected localisation Cytoplasm and ciliary tips in tissue (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A05821 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Ciliated cells (tip of cilia)); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A05821); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-TTBK2, 5 μg/mL (datasheet A05821); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Antibody staining has low consistency with RNA data (HPA tissue IHC); Isoform / epitope: 3 isoforms; no membrane segment; epitope coverage unknown (UniProt); Regulation: Testis-enhanced RNA (HPA tissue RNA). Evidence: UniProt Q6IQ55, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol and microtubules. In addition…, Datasheet A05821, PMC4256087, PMC5802059. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TTBK2/immunohistochemistry-ihc-guide.