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TTC7B / Tetratricopeptide repeat protein 7B · IHC design guide
Design Immunohistochemistry for TTC7B
Plan TTC7B paraffin IHC around membranous staining in most tissues, with prominent expression in placenta, stomach and renal tubules (HPA tissue IHC). Use 2–5 μg/ml of the IHC-validated antibody as a starting range (datasheet A12170-1), and assess membrane staining alongside the expected cytosolic pool (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TTC7B 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 | Membranous in most tissues (HPA tissue IHC); cytosolic pool expected (UniProt) | |
| Staining pattern | Membranous; abundant in placenta, stomach, renal tubules (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A12170-1) | |
| Positive control | Cerebellum+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 | Tissue staining awaits external verification (HPA tissue IHC) | |
| Regulation | EFR3-dependent membrane recruitment (UniProt) | |
| Isoform / epitope | 2 isoforms; check antibody epitope coverage (UniProt) |
Section 1
Recommended TTC7B IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A12170-1) is accompanied by one published TTC7B IHC protocol using HNSCC sections (PMC10757123).
| Sample | Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet A12170-1) |
| Fixation | Image fixative and duration unreported (datasheet A12170-1); 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 A12170-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A12170-1) |
| Primary antibody | Rabbit anti-TTC7B, 2-5 μg/ml (datasheet A12170-1) |
| Primary incubation | Overnight at 4 °C (datasheet A12170-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A12170-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TTC7B-positive staining in cells in granular layer of cerebellum (HPA tissue IHC: High). HPA tissue profile: Distinct expression in brain. Membranous expression in most tissues, highly abundant in placenta, stomach and renal tubules. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet: A12170-1); the published HNSCC protocol used microwave citrate retrieval (PMC10757123).
Section 2
What Is the Expected TTC7B Staining Pattern?
TTC7B should appear mainly at cell membranes in paraffin-section IHC, with cytosolic signal also biologically plausible because TTC7B is recruited from the cytosol to the plasma membrane (HPA: membranous expression in most tissues; UniProt Q86TV6: cytosol and EFR3-dependent membrane recruitment). Expect cell-specific staining, especially in cerebellar granular-layer cells and cerebral-cortex neuropil (HPA: High). HPA rates the tissue IHC profile Enhanced, with external verification pending (HPA: reliability).
| Crisp membrane-associated staining in the expected cells, with limited cytosolic staining. | This fits the reported membrane distribution and cytosolic pool (HPA: membranous expression in most tissues; UniProt Q86TV6: cytosol and cell membrane). Judge intensity within the identified cell population: cerebellar granular-layer cells and cortical neuropil are High, while kidney distal tubules are Medium (HPA: tissue IHC). |
| Strong, isolated nuclear staining dominates while expected membrane-associated staining is absent. | A nuclear-only pattern conflicts with the supplied localisations (HPA: plasma membrane in ICC-IF; UniProt Q86TV6: cytosol and cell membrane). Treat it as suspect staining and review a no-primary control and an independently validated antibody before assigning it to TTC7B (general IHC practice; HPA: two IHC Enhanced antibodies). |
| Adipocytes stain strongly, or staining falls on cells other than the reported positive population. | HPA reports adipocytes as Not detected in adipose tissue and breast, while identifying specific positive cell populations elsewhere (HPA: tissue IHC). Unexpected cell staining raises cross-reactivity or chromogenic detection background as possibilities; a no-primary control helps distinguish detection background from primary-antibody staining (general IHC practice). |
| Diffuse colour covers many cells and extracellular areas without a readable cellular pattern. | That distribution cannot establish the reported membrane-associated or cell-specific pattern (HPA: tissue IHC; UniProt Q86TV6: localisation). Review no-primary staining and blocking, washing and chromogen development as general IHC checks; do not score diffuse colour as TTC7B-positive solely because it appears in an HPA-positive tissue (general IHC practice). |
| A known-positive region shows no signal, including cerebellar granular-layer cells. | An absent result in a High HPA population is inconclusive until the IHC run and tissue preservation are checked (HPA: High in cerebellar granular-layer cells; general IHC practice). Verify that the expected cells are present, compare the run controls and review the antibody's documented IHC-P conditions; no target-specific fixation sensitivity is established here. |
💡Expected TTC7B appearanceCall the stain positive when membrane-associated signal maps to the expected cells, with High staining in cerebellar granular-layer cells or cortical neuropil; isolated nuclear colour or strong staining in HPA Not detected adipocytes is suspect (HPA: tissue IHC; UniProt Q86TV6: localisation).
| Compartment and cell context (HPA: tissue IHC; UniProt Q86TV6: localisation). | HPA describes membranous expression in most tissues, while UniProt also lists a cytosolic pool recruited to the membrane through EFR3. Interpret partial cytosolic signal in context; neither source supplies an IHC threshold separating membrane from cytosol. |
| Topology and processing (UniProt Q86TV6: topology and processing). | TTC7B has no annotated transmembrane segment, signal peptide or propeptide; the annotated chain spans residues 1–843. These annotations do not identify the antibody epitope or establish antigen-retrieval or fixation effects. |
| Antibody evidence (HPA: antibody validation). | HPA058363 and HPA059577 are rabbit polyclonal antibodies with IHC Enhanced status (HPA: antibodies). HPA describes the tissue profile as consistent with RNA expression, pending external verification (HPA: reliability); this supports comparison, not automatic acceptance of every stained cell. |
| Isoforms and modifications (UniProt Q86TV6: isoforms and modified residues). | Two isoforms and nine modified residues are annotated, including phosphoserines at residues 160, 202, 625, 629 and 630. The supplied record gives no antibody epitope or evidence that either feature changes IHC staining, so do not infer an isoform-specific pattern. |
| IF/ICC Q&A: What localisation should IF show? (HPA: subcellular ICC-IF). | HPA approves plasma-membrane localisation in ICC-IF and lists HPA059577 as ICC Approved (HPA: subcellular; antibodies). That is supporting localisation evidence; IF/ICC methods belong on its own guide page. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in cerebellar granular-layer cells (HPA: High). | The expected cells may be absent from the section, or the IHC run may have failed (general IHC practice); TTC7B-specific fixation sensitivity is unreported. | Confirm anatomy and cell presence, inspect run controls, and check the catalog antibody's IHC-P instructions before interpreting absence as biological (general IHC practice). |
| Signal is widespread and lacks a membrane or cell-type pattern (HPA: membranous profile). | Nonspecific primary binding, insufficient washing or chromogenic background may obscure localisation (general IHC practice). | Compare a no-primary control; review blocking, washes and development, then score only interpretable cellular staining (general IHC practice). |
| Strong nuclear-only colour appears (HPA: plasma membrane; UniProt Q86TV6: cytosol and membrane). | This compartment conflicts with the supplied localisation evidence; background or cross-reactivity is possible. | Check controls and compare with the other HPA IHC Enhanced antibody, HPA058363 or HPA059577 as applicable (HPA: antibodies; general IHC practice). |
| Adipocytes stain strongly (HPA: Not detected in adipose tissue and breast). | The observed cell type disagrees with HPA's reported pattern; cross-reactivity or endogenous detection activity may contribute (general IHC practice). | Identify the stained cells on the counterstain and inspect a no-primary control before calling them TTC7B-positive (general IHC practice). |
| Kidney distal tubules look weaker than cerebellar granular-layer cells. | That difference is compatible with HPA's Medium versus High ratings; intensity alone does not establish a technical failure (HPA: tissue IHC). | Score each tissue against its own reported cell population and level, and compare sections processed in the same run (HPA: tissue IHC; general IHC practice). |
| Cytosolic staining accompanies membrane-associated staining (UniProt Q86TV6: localisation). | UniProt reports a cytosolic pool recruited to the plasma membrane after EFR3 interaction; HPA reports a predominantly membranous tissue profile (UniProt Q86TV6; HPA: tissue IHC). | Record both compartments and their cell context. Investigate only if the cytosolic signal obscures the expected cellular pattern or also appears in controls (general IHC practice). |
Experiment evidence
IHC & IF images showing TTC7B staining
A12170-1 · IHCIHC analysis of TTC7B using anti-TTC7B antibody (A12170-1). TTC7B was detected in a paraffin-embedded section of human colorectal adenocarci
A12170-1 · IHCIHC analysis of TTC7B using anti-TTC7B antibody (A12170-1). TTC7B was detected in a paraffin-embedded section of human laryngeal squamous ce
A12170-1 · IFIF analysis of TTC7B using anti-TTC7B antibody (A12170-1). TTC7B was detected in a paraffin-embedded section of human ovarian cancer tissue.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000165914-TTC7B/tissue/cerebral+cortex#imid_16336235
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000165914-TTC7B/tissue/colon#imid_16335881
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000165914-TTC7B/tissue/liver#imid_16335874
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000165914-TTC7B/tissue/kidney#imid_16335867
HPA · ICC-IF PC-3Human Protein Atlas immunofluorescence, PC-3, antibody HPA059577; green = TTC7B, blue = nucleus, red = microtubules.
HPA · ICC-IF PC-3Human Protein Atlas immunofluorescence, PC-3, antibody HPA059577; green = TTC7B, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TTC7B IHC & IF
🧪Run cerebellum first: cells in the granular layer should stain (HPA: High). Use adipose tissue as a negative comparator, with adipocytes at background (HPA: Not detected); on the cerebellum slide, treat cells outside the granular layer as internal negatives only if their lack of staining is confirmed, since HPA does not identify an internal negative cell population there (HPA: cerebellum row).
Positive control tissue: Cerebellum (Cells in granular layer, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TTC7B in PC-3, Rh30, U2OS, with annotated localisation: Plasma membrane (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls with the same clonality as the primary, plus TTC7B-knockout tissue or cells as a biological negative where available (caption: rabbit primary; standard IHC practice). For cerebellar DAB staining, quench endogenous peroxidase and distinguish pigment from antibody signal; for IF, check tissue autofluorescence with an unstained control (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A12170-1 paraffin-section caption does not state the fixative (caption: fixative not stated). The reported IHC procedure uses heat retrieval in EDTA at pH 8.0, but its dependence on retrieval was not tested in the supplied evidence (caption: EDTA retrieval). The evidence does not establish whether frozen sections or IF are easier; HPA reports ICC-IF images in PC-3, Rh30 and U2OS, while cerebellar pigment or autofluorescence may complicate interpretation (HPA: subcellular ICC-IF images; standard IHC/IF practice).
HPA tissue IHC evidence for TTC7B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuropil | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Kidney | Distal tubules | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TTC7B IHC Tips
Troubleshoot TTC7B staining in paraffin sections by checking retrieval, compartment, tissue context and controls before interpreting chromogenic signal.
Which retrieval condition should I start with for TTC7B paraffin IHC?
Use heat-mediated EDTA retrieval at pH 8.0 for TTC7B paraffin sections (datasheet A12170-1). The selected tissue image used this condition, followed by 2 μg/ml primary antibody overnight at 4°C (caption A12170-1). If staining is weak, compare a longer heating interval or an alternative retrieval buffer on adjacent sections while keeping detection conditions fixed (standard IHC practice). Include a section processed without primary antibody to distinguish retrieval-related tissue background from antibody staining (standard IHC practice). Excessive heating can damage morphology and make cell boundaries harder to score, so assess staining alongside tissue preservation (standard IHC practice).
How should I troubleshoot fixation-related loss of TTC7B staining?
Target-specific TTC7B fixation sensitivity is unknown because the selected paraffin IHC caption does not state a fixative (caption A12170-1). Record the fixative and fixation duration for each specimen, and compare matched sections processed through the same EDTA pH 8.0 retrieval and detection steps (datasheet A12170-1; standard IHC practice). If signal varies between specimens, check preservation, staining uniformity and processing history before attributing the difference to TTC7B abundance (standard IHC practice). A no-primary control helps identify background introduced by the tissue or detection workflow (standard IHC practice). Neither the tissue staining profile nor TTC7B topology establishes a target-specific fixation effect (HPA tissue IHC; UniProt Q86TV6 topology).
Should TTC7B staining appear cytosolic or at the cell membrane?
Evaluate both cytosolic and plasma membrane staining: TTC7B is cytosolic and can be recruited to the plasma membrane through EFR3 interaction (UniProt Q86TV6 subcellular annotation). An approved plasma membrane location is reported independently, while tissue IHC describes membranous expression in most tissues (HPA subcellular; HPA tissue IHC). In chromogenic sections, inspect whether staining follows recognizable cell boundaries rather than simply outlining tissue folds or section edges (standard IHC practice). TTC7B has no annotated transmembrane segment, so membrane-associated signal does not imply an epitope inside a membrane-spanning region (UniProt Q86TV6 topology). Score cytosolic and membranous patterns separately when either contributes appreciably to the result (standard IHC practice).
Could isoform choice or epitope accessibility explain discordant TTC7B IHC staining?
TTC7B has 2 annotated isoforms, so an antibody result should be interpreted against the immunogen or epitope information for the exact reagent used (UniProt Q86TV6 isoforms; standard IHC practice). The supplied caption identifies A12170-1 staining conditions but does not locate its epitope or establish isoform coverage (caption A12170-1). If sections disagree, compare the documented epitope with the isoform sequences before calling one specimen negative (standard IHC practice). TTC7B also has annotated phosphoserines, including positions 160, 202 and 625, but their effect on this antibody is unreported (UniProt Q86TV6 modified residues). Test retrieval changes on adjacent sections without assuming that phosphorylation caused a staining difference (standard IHC practice).
How can I check a TTC7B pattern by multiplex immunofluorescence?
Choose a marker for the cell population being evaluated, then inspect TTC7B and marker signals in separate channels as well as the merged image (standard IF practice). Brain signal is reported in cerebellar granular-layer cells and cerebral-cortex neuropil, while kidney distal tubules are reported at a medium level; select the marker to fit the specimen (HPA tissue IHC). Choose fluorophores and exposure settings after checking the section's autofluorescence and single-label controls (standard IF practice). Because TTC7B can be cytosolic or plasma membrane associated and has no transmembrane segment, set permeabilisation according to the antibody's documented epitope accessibility rather than assuming a membrane-spanning target (UniProt Q86TV6 subcellular annotation and topology; standard IF practice).
What should I check when TTC7B DAB staining looks diffuse?
Compare the stained section with a no-primary control and inspect whether diffuse colour follows tissue edges, folds or damaged areas (standard IHC practice). The selected paraffin example used 10% goat serum blocking, a peroxidase-conjugated secondary antibody and DAB development (caption A12170-1). Check the peroxidase block and secondary-only background within the local chromogenic workflow, because endogenous enzyme activity or nonspecific detection can produce DAB signal (standard IHC practice). If background remains high, adjust primary concentration or incubation while retaining the documented EDTA pH 8.0 retrieval as the starting condition (datasheet A12170-1; standard IHC practice). Judge improvement by preserved cell morphology and compartment-specific staining, not by darker colour alone (standard IHC practice).
How should I quantify TTC7B staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because TTC7B may appear in cytosol or at the plasma membrane (UniProt Q86TV6 subcellular annotation). For cellular staining, record the percentage of positive cells and an H-score from intensity categories applied consistently across sections (standard IHC practice). For a spatial compartment such as neuropil, use stained area or optical density per mm² of evaluable tissue instead of forcing a cell count (HPA: high in cerebral-cortex neuropil; standard IHC practice). Normalize to the same eligible tissue area or cell population, exclude folds and necrosis, and hold imaging and DAB development settings constant (standard IHC practice). Report cytosolic and membranous scores separately when both are present (standard IHC practice).
When is a TTC7B-positive IHC result convincing?
A convincing result has reproducible staining in an interpretable cell population, with cytosolic or plasma membrane distribution compatible with TTC7B annotation (UniProt Q86TV6 subcellular annotation; standard IHC practice). Tissue context matters: HPA reports high signal in cerebellar granular-layer cells and medium signal in kidney distal tubules and lung macrophages (HPA tissue IHC). Treat nuclear-only colour, section-edge accentuation, necrotic deposits or signal also present without primary antibody as reasons to investigate artefact (UniProt Q86TV6 subcellular annotation; standard IHC practice). Review peroxidase blocking if DAB appears broadly in cells or tissue structures outside the expected pattern (standard IHC practice). A negative region is interpretable only when morphology and assay controls support adequate detection (standard IHC practice).
Boster reagents
Best TTC7B / Tetratricopeptide repeat protein 7B IHC Antibodies
A12170-1 has real IHC data from human paraffin sections and IF data from a human paraffin section; its listed reactivity includes human, mouse and rat (catalog applications, reactivity and image captions).
Real IHC data
Anti-TTC7B Antibody ®
Cat # A12170-1
A12170-1 is the sole SKU and has IHC images from human colorectal adenocarcinoma, laryngeal squamous cell carcinoma, liver cancer and breast cancer paraffin sections (catalog IHC image captions). Its IF image shows a human ovarian cancer paraffin section; the catalog lists human, mouse and rat reactivity (catalog IF image caption and reactivity).
Which to pick: For tissue IHC, choose A12170-1: its paraffin section image used EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody; the fixative is unreported (A12170-1 IHC image caption). For IF, A12170-1 has a human paraffin section image at 5 μg/ml, while ICC validation is unreported (A12170-1 IF image caption and application list). For cross-species work, A12170-1 lists human, mouse and rat reactivity, but the application-specific dilution text lists IHC for human and rat and IF for human; clonality is unreported (A12170-1 catalog reactivity, dilution text and clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q86TV6 (TTC7B_HUMAN, Tetratricopeptide repeat protein 7B).
- Human Protein Atlas. TTC7B tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. TTC7B subcellular location (ICC-IF): Localized to the plasma membrane..
- Human Protein Atlas. TTC7B antibody validation summary (2 antibodies).
- TTC7B is a new prognostic biomarker in head and neck squamous cell carcinoma linked to immune infiltration and ferroptosis. Cancer medicine 2023 — PMC10757123.
- GIPC1 Restrains the Progression and Chemoresistance of Colorectal Cancer by Regulating TTC7B/mTOR/NF-κB Axis. International journal of biological sciences 2026 — PMC12781073.
- PubMed PMID:12508121 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TTC7B (Tetratricopeptide repeat protein 7B). Expected localisation Membranous in most tissues (HPA tissue IHC); cytosolic pool expected (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A12170-1). Validated IHC antibody: A12170-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Cells in granular layer); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet A12170-1); 2. Fixation: Image fixative and duration unreported (datasheet A12170-1); 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 A12170-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A12170-1); 8. Primary antibody: Rabbit anti-TTC7B, 2-5 μg/ml (datasheet A12170-1); 9. Primary incubation: Overnight at 4 °C (datasheet A12170-1). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Tissue staining awaits external verification (HPA tissue IHC); Isoform / epitope: 2 isoforms; check antibody epitope coverage (UniProt); Regulation: EFR3-dependent membrane recruitment (UniProt). Evidence: UniProt Q86TV6, HPA tissue IHC · Enhanced, HPA ICC-IF · plasma membrane., Datasheet A12170-1, PMC10757123. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TTC7B/immunohistochemistry-ihc-guide.