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CCDC134 / Coiled-coil domain-containing protein 134 · IHC design guide
Design Immunohistochemistry for CCDC134
Plan chromogenic IHC on paraffin sections for CCDC134 using the IHC-validated antibody at a starting concentration of 2.5 μg/mL (datasheet). This guide highlights the cytoplasmic tissue pattern and respiratory epithelial staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CCDC134 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 in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in respiratory epithelial 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 | Bone marrow+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); target-specific effects are unknown. | |
| Caveat | Secretion can make tissue protein and RNA patterns differ (HPA tissue IHC) | |
| Regulation | Higher expression in activated CD8+ T cells (UniProt) | |
| Isoform / epitope | 0 isoforms; mature chain starts at residue 23 after signal peptide cleavage (UniProt) |
Section 1
Recommended CCDC134 IHC & IF Protocols
The catalog antibody’s IHC-P protocol and one published CCDC134 IHC procedure (PMC5746300) provide starting conditions for paraffin sections.
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A14619); 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-CCDC134, 2.5 μg/mL (datasheet A14619) |
| 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 | CCDC134-positive staining in respiratory epithelial cells of bronchus (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min (page retrieval rule); compare citrate steaming for 3 min (PMC5746300).
Section 2
What Is the Expected CCDC134 Staining Pattern?
CCDC134 is mainly in the endoplasmic reticulum lumen and has no transmembrane segment; it is also reported as secreted, cytoplasmic and nuclear (UniProt Q9H6E4). In paraffin tissue sections, expect predominantly cytoplasmic staining in many tissues (HPA: IHC profile, Approved). Respiratory epithelial cells in bronchus and nasopharynx are useful high-staining examples (HPA: High). Secretion means tissue RNA and protein locations may differ (HPA: reliability description).
| Cytoplasmic staining in bronchial or nasopharyngeal respiratory epithelial cells. | This matches the reported tissue profile and high staining level (HPA: cytoplasmic expression in most tissues; High in these cells). Score the relevant epithelial cells separately from surrounding tissue (general IHC practice). |
| Predominantly nuclear staining across an otherwise routine tissue section. | Treat this as a compartment mismatch against the usual cytoplasmic IHC profile (HPA: tissue IHC). Nuclear accumulation is reported after UV irradiation (UniProt Q9H6E4), so context matters; check the control and staining distribution before interpreting it. |
| Strong staining in esophageal or oral squamous epithelial cells. | These cell types were not detected in the cited tissue IHC observations (HPA: esophagus; oral mucosa). Unexpected staining warrants a check for cross-reactivity or endogenous detection activity (general IHC practice); it alone does not identify the cause. |
| Uniform color over cells, extracellular space and the negative control. | A diffuse field does not resemble the cell-associated cytoplasmic profile (HPA: tissue IHC). Consider nonspecific background or detection chemistry and compare controls before assigning CCDC134 positivity (general IHC practice). |
| No signal in respiratory epithelium of a bronchus or nasopharynx control. | Both are reported as high-staining sites (HPA: bronchus; nasopharynx). Review tissue integrity, retrieval, antibody incubation and chromogenic detection as general IHC checks; a failed control prevents a confident negative call in the test section. |
💡Expected CCDC134 appearanceCall a positive result when respiratory epithelial cells show clear, predominantly cytoplasmic staining, with strong signal plausible in bronchus or nasopharynx (HPA: IHC profile; High); widespread nuclear-only or uniform extracellular color is a pattern mismatch requiring control review (HPA: IHC profile; general IHC practice).
| Secretion and tissue distribution | CCDC134 is secreted and lacks a transmembrane segment (UniProt Q9H6E4). Its protein location need not follow tissue RNA location (HPA: reliability description); assess stained cell types directly. |
| Intracellular compartment | The ER lumen is the main location reported by UniProt, while HPA tissue IHC describes a cytoplasmic pattern (UniProt Q9H6E4; HPA: tissue IHC). Routine chromogenic IHC may show this as cytoplasmic staining. |
| Processing and glycosylation | The annotated signal peptide spans residues 1–22, the mature chain spans 23–229, and one glycosylation site is annotated at 148 (UniProt Q9H6E4). These annotations do not establish epitope accessibility after retrieval. |
| Evidence across assays | IHC is Approved for HPA003936; ICC is Supported for HPA075348 (HPA: antibodies). IF/ICC Q&A: should the image match paraffin IHC exactly? No: HPA reports cytosol with additional vesicles in ICC-IF, while tissue IHC is broadly cytoplasmic (HPA: subcellular; tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive control is blank. | Bronchial and nasopharyngeal respiratory epithelial cells are reported High (HPA: tissue IHC); a blank control suggests an assay or specimen problem, without identifying which. | Inspect section quality and confirm retrieval, antibody incubation and detection steps (general IHC practice). Resolve the control before scoring test tissue. |
| Signal appears mainly in nuclei. | This differs from the usual cytoplasmic IHC profile (HPA: tissue IHC), although UV-associated nuclear accumulation is reported (UniProt Q9H6E4). | Check exposure context if known, then compare with the positive and negative controls and review whether nuclear color is cell-specific (general IHC practice). |
| Unexpected squamous epithelial staining dominates. | Esophageal and oral squamous epithelial cells were not detected in HPA tissue IHC (HPA: esophagus; oral mucosa); cross-reactivity or endogenous activity are possibilities (general IHC practice). | Compare a negative control and the expected respiratory epithelial positive control; investigate detection background before calling this target-specific staining (general IHC practice). |
| Diffuse color obscures cell boundaries. | The reported pattern is cytoplasmic expression in most tissues (HPA: tissue IHC); widespread color may reflect nonspecific staining or detection background (general IHC practice). | Review blocking, washes, antibody concentration and chromogen development against controls (general IHC practice). Reassess only when cellular staining is distinguishable. |
| Low or absent signal in one tissue is treated as a universal negative. | Expression varies by cell type: liver hepatocytes are Low, while respiratory epithelial cells are High (HPA: tissue IHC). A single tissue does not establish assay sensitivity. | Include a documented high-staining respiratory epithelial control and score the relevant cell population, not the whole section (HPA: tissue IHC; general IHC practice). |
| IF/ICC appears vesicular while IHC appears broadly cytoplasmic. | HPA reports supported cytosolic and additional approved vesicular ICC-IF localization, alongside a cytoplasmic tissue IHC profile (HPA: subcellular; tissue IHC). | Interpret each assay against its own reported pattern and controls; do not use the ICC-IF image alone to redefine a paraffin IHC positive call (HPA: subcellular; tissue IHC). |
Experiment evidence
IHC & IF images showing CCDC134 staining
A14619 · IHCImmunohistochemistry of CCDC134 in human brain tissue with CCDC134 antibody at 2.5 μg/mL.
A14619 · IFImmunofluorescence of CDCC134 in human brain tissue with CDCC134 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000100147-CCDC134/tissue/cerebral+cortex#imid_1326332
HPA · NasopharynxHuman Protein Atlas IHC, Nasopharynx. Source: https://www.proteinatlas.org/ENSG00000100147-CCDC134/tissue/nasopharynx#imid_1326321
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000100147-CCDC134/tissue/colon#imid_1326258
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000100147-CCDC134/tissue/liver#imid_1326259
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA075348; green = CCDC134, blue = nucleus, red = microtubules.
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA075348; green = CCDC134, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CCDC134 IHC & IF
🧪Run bronchus first: respiratory epithelial cells should stain (HPA: High in bronchus respiratory epithelial cells). Use esophageal squamous epithelium as a negative tissue (HPA: Not detected in esophagus squamous epithelial cells); internal negative cells on the bronchus slide should show counterstain and background only, without specific chromogen (standard IHC control criterion).
Positive control tissue: Bronchus (Respiratory epithelial cells, HPA High)
Negative control tissue: Bone marrow (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CCDC134 in HeLa, SiHa, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary/secondary-only and an isotype control matched to the primary antibody’s host species and clonality, plus CCDC134 knockout tissue or a validated peptide-block control (standard IHC controls). Block endogenous peroxidase for chromogenic detection in bronchus, and check for background staining in the no-primary slide (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; the fixative is also unreported in the A14619 tissue-IHC caption (selected IHC caption). The evidence does not establish whether frozen sections or IF are easier for CCDC134 (supplied application evidence). In bronchus, distinguish cellular staining from chromogen or antibody retained in luminal mucus (standard IHC artefact check).
HPA tissue IHC evidence for CCDC134
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Secreted protein, tissue location of RNA and protein is expected to differ. Caution, Splice and/or transcript discrepancy exists.). 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 | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CCDC134 IHC Tips
Troubleshoot CCDC134 staining in paraffin sections by checking retrieval, compartment, cell type and controls before comparing chromogenic signal across samples.
What retrieval should I start with for CCDC134 in paraffin sections?
Use Tris-EDTA pH 9.0 heat-induced epitope retrieval at 95–98 °C for 20 min (page retrieval setting). Cool sections consistently, then compare staining with a no-primary control and a known positive tissue processed in the same run (standard IHC practice; HPA: high in bronchial respiratory epithelial cells). If staining remains weak, test citrate pH 6.0 as a fallback on adjacent sections while holding antibody concentration and detection constant (standard IHC optimisation). Score epithelial cytoplasm separately from nuclei because tissue IHC is predominantly cytoplasmic, whereas nuclear accumulation is reported after UV irradiation (HPA: cytoplasmic expression in most tissues; UniProt Q9H6E4: UV-responsive nuclear accumulation).
Could fixation explain weak or patchy CCDC134 staining?
Target-specific fixation sensitivity for CCDC134 is unknown from the supplied evidence; the selected tissue-IHC caption does not state its fixative (A14619 tissue-IHC caption). For paraffin IHC, document fixative, fixation duration, section age and retrieval conditions before comparing cases, because these variables can affect epitope accessibility (standard IHC practice). Run a known positive tissue beside the problem section and use the same retrieval and detection sequence to distinguish a run-wide failure from a specimen-specific problem (standard IHC practice; HPA: high in bronchial respiratory epithelial cells). Do not treat the caption’s 2.5 µg/mL antibody concentration as evidence that any particular fixation condition was validated (A14619 tissue-IHC caption).
Should I expect cytoplasmic, ER-like or nuclear CCDC134 staining?
Assess cytoplasmic signal first: tissue IHC reports cytoplasmic expression in most tissues, while UniProt says CCDC134 mainly localizes to the ER (HPA: tissue profile; UniProt Q9H6E4: subcellular location). A fine intracellular pattern is plausible, but chromogenic IHC alone cannot establish ER residence; compare morphology and a separately validated compartment marker if that distinction matters (UniProt Q9H6E4: ER localisation; standard IHC practice). Record nuclear staining separately because nuclear accumulation was reported in response to UV irradiation, not as a universal tissue pattern (UniProt Q9H6E4: PubMed:22644376). Diffuse extracellular staining requires cautious interpretation because CCDC134 also has a secreted form (UniProt Q9H6E4: secreted localisation).
How do processing and epitope placement affect interpretation of CCDC134 staining?
CCDC134 has a 1–22 signal peptide and a 23–229 annotated chain, with no transmembrane segment (UniProt Q9H6E4: processing and topology). Check the antibody’s stated immunogen or epitope before interpreting negative staining: recognition of the signal peptide could differ from recognition of the mature chain (UniProt Q9H6E4: processing; standard antibody interpretation). One glycosylation site is annotated at residue 148, but the supplied evidence does not establish whether it affects this antibody’s tissue staining (UniProt Q9H6E4: glycosylation). No isoforms are listed in the supplied record; avoid assigning a staining pattern to a particular isoform, especially given the reported transcript discrepancy (UniProt Q9H6E4: isoforms; HPA: reliability note).
How should I investigate CCDC134 localisation by IF alongside chromogenic IHC?
Use IF/ICC as a separate localisation experiment rather than treating its image patterns as validation of the paraffin IHC workflow (HPA: subcellular IF images; A14619 tissue-IHC caption). Multiplex CCDC134 with a marker for the cell type under study; bronchial respiratory epithelial cells offer a tissue-IHC reference with high reported staining (HPA: high in bronchial respiratory epithelial cells; standard IF practice). Choose fluorophores after checking each specimen’s autofluorescence and include single-stain controls to assess channel bleed-through (standard IF practice). For an intracellular ER-lumen target, optimise permeabilisation so antibody can reach the luminal epitope, and interpret cytosolic or vesicular signal against the reported IF pattern (UniProt Q9H6E4: ER-lumen localisation; HPA: cytosol supported, vesicles approved).
How can I reduce widespread brown signal without losing CCDC134 staining?
Compare the stained section with no-primary and detection-only controls to identify signal arising from the chromogenic detection workflow (standard IHC practice). Include an endogenous peroxidase block when using HRP and DAB, and optimise blocking, washes, primary concentration and development time one variable at a time (standard IHC practice). The selected image caption reports 2.5 µg/mL for antibody A14619 in human brain tissue, but gives no fixation details or transferable optimum for other specimens (A14619 tissue-IHC caption). Evaluate background against the expected predominance of cytoplasmic tissue staining, while allowing for cell-type differences and secreted CCDC134 (HPA: tissue profile; UniProt Q9H6E4: secreted localisation).
How should I quantify CCDC134 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then apply the same threshold and sampling rule to every section (standard IHC quantification; HPA: predominantly cytoplasmic tissue staining). An H-score combines the percentage of cells at each intensity from 0 to 3; alternatively report percentage positive cells or positive-cell density per mm² (standard IHC quantification). Normalise to the number or area of eligible cells in the same tissue compartment, and exclude folds, edges and necrotic regions consistently (standard IHC quantification). Report nuclear and cytoplasmic results separately if both occur, because UV-responsive nuclear accumulation and predominant tissue cytoplasmic staining have distinct evidence (UniProt Q9H6E4: PubMed:22644376; HPA: tissue profile).
When is an unexpected CCDC134-positive area likely to be artefact?
Treat intracellular cytoplasmic staining in the expected cell population as more credible when it reproduces across sections and the controls behave as expected (HPA: cytoplasmic expression in most tissues; standard IHC practice). For example, respiratory epithelial cells have high reported tissue-IHC signal, whereas germinal center cells in lymph node were not detected (HPA: bronchus and lymph node profiles). Isolated nuclear staining merits separate review because nuclear accumulation is linked to UV irradiation in the supplied record (UniProt Q9H6E4: PubMed:22644376). Check abrupt section-edge staining, necrotic deposits and signal retained in no-primary or peroxidase controls before calling a focus positive; secreted CCDC134 also makes extracellular deposits harder to assign to their source cell (standard IHC practice; UniProt Q9H6E4: secreted localisation).
Boster reagents
Best CCDC134 / Coiled-coil domain-containing protein 134 IHC Antibodies
A14619 has real IHC data and an IF image in human brain tissue (catalog: figure tag and image captions). Its listed reactivity covers human, mouse and rat (catalog: reactivity).
Real IHC data
Anti-CCDC134 Antibody
Cat # A14619
A14619 is listed for IHC-P and IF, with human, mouse and rat reactivity (catalog: applications and reactivity). Its human brain images show IHC at 2.5 μg/mL and IF at 20 μg/mL (A14619: IHC and IF image captions).
Which to pick: For paraffin-section IHC, choose A14619: IHC-P is listed, and its own IHC image shows human brain staining at 2.5 μg/mL (catalog: applications; A14619 IHC image caption); the fixative is unreported (A14619 IHC image caption). For IF, choose A14619, whose human brain IF image uses 20 μg/mL; separate ICC validation is not listed (A14619 IF image caption; catalog: applications). For mouse or rat samples, A14619 has listed reactivity, but both supplied tissue images are human; clonality is unspecified (catalog: reactivity and clone; A14619 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9H6E4 (CC134_HUMAN, Coiled-coil domain-containing protein 134).
- Human Protein Atlas. CCDC134 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. CCDC134 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to vesicles..
- Human Protein Atlas. CCDC134 antibody validation summary (2 antibodies).
- CCDC134 serves a crucial role in embryonic development. International journal of molecular medicine 2018 — PMC5746300.
- PubMed PMID:15461802 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:10591208 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CCDC134 (Coiled-coil domain-containing protein 134). Expected localisation Cytoplasmic in most tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A14619 at 2.5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Respiratory epithelial cells); negative Bone marrow. 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 A14619); 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-CCDC134, 2.5 μg/mL (datasheet A14619); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); target-specific effects are unknown.; Caveat: Secretion can make tissue protein and RNA patterns differ (HPA tissue IHC); Isoform / epitope: 0 isoforms; mature chain starts at residue 23 after signal peptide cleavage (UniProt); Regulation: Higher expression in activated CD8+ T cells (UniProt). Evidence: UniProt Q9H6E4, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to vesicles., Datasheet A14619, PMC5746300. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CCDC134/immunohistochemistry-ihc-guide.