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CCT8 / T-complex protein 1 subunit theta · IHC design guide
Design Immunohistochemistry for CCT8
Plan chromogenic CCT8 IHC in paraffin sections around the reported cytoplasmic tissue pattern (HPA tissue IHC). Start the catalog antibody at 0.5–1 μg/mL (datasheet PB9993), and compare staining by cell type, including high staining in testis spermatogonia and undetected staining in smooth muscle cells (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CCT8 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 tissue staining (HPA tissue IHC) | |
| Staining pattern | General cytoplasmic staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9993) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Smooth muscle+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9993) | |
| Caveat | Smooth muscle cells may be unstained (HPA tissue IHC) | |
| Regulation | No specific expression regulator annotated (UniProt) | |
| Isoform / epitope | 3 isoforms; check epitope coverage for each (UniProt) |
Section 1
Recommended CCT8 IHC & IF Protocols
The catalog antibody protocol (datasheet PB9993) appears alongside published CCT8 IHC protocols for colorectal cancer and lung adenocarcinoma tissues (PMC8642402; PMC10622983).
| Sample | Paraffin-embedded mouse kidney tissue; fixative not specified (datasheet PB9993) |
| Fixation | Image fixative and duration unreported (datasheet PB9993); 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 PB9993); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9993) |
| Primary antibody | Rabbit anti-CCT8, 0.5-1μg/ml (datasheet PB9993) |
| Primary incubation | Overnight at 4 °C (datasheet PB9993) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9993) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CCT8-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet PB9993). The lung tissue protocol used microwave citrate pH 6.0 (PMC10622983).
Section 2
What Is the Expected CCT8 Staining Pattern?
CCT8 is a cytoplasmic TRiC chaperonin subunit with reported centrosome and cilium basal body localisation; it has no transmembrane segment (UniProt P50990). In paraffin section IHC, expect mainly cytoplasmic staining in the cell populations reported as high by HPA, including adrenal glandular cells, bronchial respiratory epithelium and placental cytotrophoblasts (HPA: tissue IHC). HPA rates its tissue IHC evidence Enhanced, with medium consistency between staining and RNA expression (HPA: tissue IHC).
| Clear cytoplasmic stain in adrenal glandular cells, bronchial respiratory epithelial cells or placental cytotrophoblasts (HPA: High in each). | This fits the reported general cytoplasmic pattern (HPA: tissue IHC) and CCT8 localisation (UniProt P50990). Compare cells within the same section: HPA reports cell-specific levels, so an entire tissue need not stain uniformly (HPA: tissue IHC). |
| Predominantly nuclear staining in tissue IHC, with little cytoplasmic signal (HPA: general cytoplasmic tissue profile). | Treat this as a compartment mismatch requiring validation. HPA's uncertain nucleoplasmic ICC-IF location cannot establish a predominantly nuclear pattern in paraffin tissue IHC (HPA: subcellular ICC-IF; HPA: tissue IHC). |
| Strong stain confined to smooth muscle cells or chondrocytes (HPA: Not detected in both). | This conflicts with HPA's observed tissue IHC pattern and warrants checks for antibody cross-reactivity or endogenous detection activity. “Not detected” describes HPA observations; it is not proof that every specimen must be negative (HPA: tissue IHC). |
| Broad colour deposit across cells and surrounding tissue, obscuring cytoplasmic boundaries (HPA: general cytoplasmic tissue profile). | Interpret as background until controls clarify it. In chromogenic IHC, excess primary antibody, inadequate blocking or residual endogenous detection activity can produce nonspecific colour; those are general workflow possibilities, not reported CCT8-specific effects. |
| No cytoplasmic stain in adrenal glandular cells or bronchial respiratory epithelium (HPA: High in both). | A known-positive cell population failing to stain makes a negative result elsewhere difficult to interpret. Check section quality, retrieval and detection controls before inferring absent CCT8; HPA's Enhanced rating does not guarantee every antibody and run (HPA: tissue IHC). |
💡Expected CCT8 appearanceCall positive when the expected cells show clear, mainly cytoplasmic signal—for example, high staining in placental cytotrophoblasts (HPA: High; general cytoplasmic profile); predominant nuclear colour or strong staining in HPA-negative smooth muscle cells is suspect (HPA: tissue IHC).
| Tissue and cell selection | HPA reports high IHC staining in hippocampal neurons, ovarian follicle cells, stomach glandular cells and testicular spermatogonia, alongside other high populations; smooth muscle cells and chondrocytes were not detected (HPA: tissue IHC). Use the named cell type when judging a section. |
| Antibody evidence | Three listed antibodies have Enhanced IHC validation (HPA: HPA018520, HPA021051, HPA029426). The tissue profile also has medium staining-to-RNA consistency (HPA: tissue IHC). These ratings support a comparison pattern but do not establish performance for an unlisted antibody or a particular retrieval condition. |
| Protein organisation and variants | CCT8 has no transmembrane segment or signal peptide, and UniProt lists three isoforms (UniProt P50990). Cytoplasmic staining is consistent with its localisation (UniProt P50990; HPA: tissue IHC). The supplied records do not map antibody epitopes to isoforms, so isoform-specific IHC claims are unsupported. |
| IF/ICC: should nuclear or sperm staining change the tissue IHC call? | HPA lists nucleoplasm as uncertain, cytosol as supported, and sperm connecting and end pieces as approved ICC-IF locations (HPA: subcellular ICC-IF). These image-based locations answer the IF/ICC question; they do not replace the general cytoplasmic tissue IHC profile (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive cells show no stain (HPA: High in adrenal glandular cells). | The IHC run may have failed, or retrieval and detection settings may be unsuitable; no CCT8-specific fixation sensitivity is reported in the supplied sources. | Confirm that the named cells are present, then review the run's positive control, retrieval record, antibody incubation and detection steps. Treat any adjustment as general IHC optimisation. |
| Signal is weak in a chosen comparison tissue (HPA: Low in oral mucosal squamous epithelium). | HPA reports low staining in that cell population (HPA: tissue IHC); a faint result may reflect the expected reference level. | Compare with a reported high population, such as stomach glandular cells (HPA: High), before changing staining conditions. Score the specified cells rather than the whole tissue. |
| Nuclear stain dominates a paraffin section (HPA: general cytoplasmic tissue profile). | The tissue result may reflect nonspecific stain; HPA's nucleoplasmic ICC-IF assignment is uncertain (HPA: subcellular ICC-IF). | Recheck controls and whether cytoplasmic signal is present in a reported high cell population (HPA: tissue IHC). Do not use the uncertain ICC-IF assignment alone to call nuclear tissue IHC positive. |
| Smooth muscle cells or chondrocytes stain strongly (HPA: Not detected in both). | Possible cross-reactivity or endogenous chromogenic detection activity; this is a general IHC differential, not a demonstrated CCT8 mechanism. | Inspect a no-primary control and the detection-blocking steps. Compare the staining distribution with HPA's named positive and negative cell populations (HPA: tissue IHC). |
| Diffuse chromogen obscures cell borders (HPA: general cytoplasmic tissue profile). | Excess primary antibody, insufficient blocking or incomplete washing may raise general IHC background; no target-specific cause is established. | Check the no-primary control, blocking and wash steps, then optimise primary antibody concentration within the chosen assay. Interpret CCT8 only when cell-associated cytoplasmic stain can be distinguished. |
| Different antibodies give conflicting tissue patterns (HPA: three Enhanced IHC antibodies). | Validation status does not make all reagents equivalent; epitope coverage across the three UniProt isoforms is not supplied (UniProt P50990; HPA: antibodies). | Compare each reagent in the same reported high and not-detected cell populations (HPA: tissue IHC). Review its own validation and controls before attributing differences to isoforms or tissue biology. |
Experiment evidence
IHC & IF images showing CCT8 staining
PB9993 · IHCIHC analysis of CCT8 using anti-CCT8 antibody (PB9993). CCT8 was detected in a paraffin-embedded section of mouse kidney tissue. Heat mediat
PB9993 · IHCIHC analysis of CCT8 using anti-CCT8 antibody (PB9993). CCT8 was detected in a paraffin-embedded section of rat testis tissue. Heat mediated
PB9993 · IFIF analysis of CCT8 using anti-CCT8 antibody (PB9993). CCT8 was detected in an immunocytochemical section of A549 cells. Enzyme antigen retr
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000156261-CCT8/tissue/cerebral+cortex#imid_7613256
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000156261-CCT8/tissue/colon#imid_7613190
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000156261-CCT8/tissue/liver#imid_7613191
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000156261-CCT8/tissue/kidney#imid_7613192
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018520; green = CCT8, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018520; green = CCT8, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CCT8 IHC & IF
🧪Run adrenal gland first and look for staining in glandular cells (HPA: High in adrenal glandular cells). Use smooth muscle as the negative tissue (HPA: Not detected in smooth muscle cells); on the adrenal slide, any unstained neighboring cells can serve as an internal background reference, but their CCT8-negative status is unverified.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Smooth muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CCT8 in A-431, U-251MG, U2OS, Sperm, with annotated localisation: Nucleoplasm (uncertain), Connecting piece (approved), End piece (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control for the rabbit primary antibody (PB9993 caption: rabbit anti-CCT8). Use CCT8-knockout material as a biological negative where available. Quench endogenous peroxidase and check for endogenous biotin signal, since the illustrated chromogenic workflow uses biotinylated secondary antibody, SABC and DAB (PB9993 caption).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative used for the illustrated paraffin section is unreported (PB9993 caption). The illustrated IHC result used heat retrieval in EDTA at pH 8.0, so start with that condition and verify its effect in the chosen tissue; the caption does not establish that retrieval is required in every specimen (PB9993 caption). The supplied evidence does not establish whether frozen sections or IF are easier; for adrenal sections, check pigment and other background against the no-primary control (HPA: High in adrenal glandular cells).
HPA tissue IHC evidence for CCT8
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | High | Protein (IHC) | HPA → |
| Ovary | Follicle cells | High | Protein (IHC) | HPA → |
| Placenta | Cytotrophoblasts | High | Protein (IHC) | HPA → |
Section 3
Advanced CCT8 IHC Tips
Troubleshoot CCT8 chromogenic IHC in paraffin sections by checking retrieval, cell compartment, detection background, and cell specific scoring.
What retrieval should I use when CCT8 staining is weak in paraffin sections?
Use heat mediated retrieval in EDTA at pH 8.0 for paraffin section CCT8 IHC (datasheet PB9993). The selected mouse kidney image used this retrieval before an overnight primary incubation at 4°C, but reports no retrieval duration or temperature (caption PB9993). Keep section thickness, heating, cooling, and detection conditions matched when comparing retrieval runs (standard IHC practice). If staining remains weak, test a different retrieval condition on adjacent sections as a fallback, while retaining the EDTA condition as the reference (standard IHC practice). Score cytoplasmic signal in intact cells against background on a primary omitted control (UniProt P50990 localisation; standard IHC practice).
Could fixation explain weak or uneven CCT8 staining?
The selected paraffin section caption does not state a fixative, so target specific fixation sensitivity is unknown (caption PB9993). Record the actual fixative, fixation interval, tissue thickness, and processing history for every specimen before comparing staining (standard IHC practice). If those variables differ, stain adjacent sections with the same EDTA pH 8.0 retrieval and antibody conditions to identify a reproducible processing effect (datasheet PB9993; standard IHC practice). Compare intact cells within equivalent tissue regions and include a primary omitted control to reveal detection background (standard IHC practice). Do not assign a fixation mechanism to CCT8 from its annotated localisation or modified residues (UniProt P50990; standard IHC practice).
Which compartments should count as plausible CCT8 staining?
Expect mainly cytoplasmic staining in tissue IHC, consistent with the reported general cytoplasmic profile (HPA tissue IHC). CCT8 is also annotated at the centrosome and cilium basal body, so a discrete pericentrosomal signal can be plausible if its morphology is convincing (UniProt P50990 localisation). The subcellular atlas additionally reports supported cytosol and uncertain nucleoplasm, alongside approved sperm structures; those observations should not make isolated nuclear staining a default positive call in tissue sections (HPA subcellular). Evaluate nuclear only or diffuse extracellular color against matched controls and adjacent morphology (standard IHC practice). Record cytoplasmic and nuclear signals separately when they coexist (standard IHC practice).
Can an epitope or isoform difference explain discordant CCT8 staining?
CCT8 has 3 annotated isoforms, but the supplied antibody caption does not identify its epitope or establish isoform coverage (UniProt P50990 isoforms; caption PB9993). Map the immunogen or epitope against each isoform before interpreting differences between specimens as isoform specific expression (standard IHC practice). CCT8 also has annotated modifications, including phosphorylation at residues 23, 30, and 162; no supplied evidence shows that these alter this antibody’s binding (UniProt P50990 modified residues; caption PB9993). Compare staining with an independently validated antibody to a distinct epitope, if available, under matched retrieval and detection conditions (standard IHC practice). Treat any discrepancy as unresolved until epitope identity and specificity are checked (standard IHC practice).
How should I investigate CCT8 localisation with multiplex IF?
Use IF as a separate validation experiment for a puzzling chromogenic IHC pattern, because the supplied PB9993 tissue caption documents chromogenic paraffin IHC (caption PB9993). Pair CCT8 with a validated marker of the cell population under study, such as a marker identifying cytotrophoblasts when examining placenta (HPA tissue IHC; standard IF practice). Check tissue autofluorescence in unstained controls, then choose spectrally separated fluorophores, favoring a longer wavelength CCT8 channel if background is strong (standard IF practice). CCT8 has no annotated transmembrane segment and has cytoplasmic localisation, so assess permeabilisation sufficient to access its intracellular epitope (UniProt P50990 topology and localisation; standard IF practice). Confirm the antibody’s IF performance independently before comparing IF signal with IHC scores (standard IF practice).
How can I separate weak CCT8 staining from chromogenic background?
The selected image used biotinylated goat anti rabbit secondary antibody, streptavidin biotin complex detection, and DAB (caption PB9993). With that workflow, run primary omitted and secondary omitted controls to locate nonspecific secondary signal and detection reagent background (standard IHC practice). Check endogenous peroxidase blocking and, if needed for this detection system, endogenous biotin blocking on matched sections (standard IHC practice). Compare suspected CCT8 staining with intact cell boundaries: the expected general tissue profile is cytoplasmic (HPA tissue IHC). If diffuse color persists in controls, adjust blocking, washing, or detection exposure before changing the primary concentration; the selected primary concentration was 1 μg/ml (caption PB9993; standard IHC practice).
How should I quantify heterogeneous CCT8 staining across tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define a cell population and tissue region before scoring, then quantify cytoplasmic DAB within viable, morphologically intact cells (HPA tissue IHC; standard IHC practice). Report the percentage of positive cells and staining intensity, or calculate an H score by summing the percentages at intensity 1, 2, and 3 multiplied by their grades, giving a 0–300 scale (standard IHC practice). Normalize cell counts to the number of eligible cells, or positive cell density to analyzed area in mm², using the same denominator across specimens (standard IHC practice). Set thresholds with matched negative controls and apply them consistently (standard IHC practice). Keep nuclear and cytoplasmic scores separate if both appear (standard IHC practice).
When is apparent CCT8 positivity more likely an artefact?
A credible tissue signal is intracellular and predominantly cytoplasmic, consistent with the reported general tissue IHC pattern and CCT8 localisation (HPA tissue IHC; UniProt P50990 localisation). Strong color confined to section edges, folds, necrotic areas, or extracellular material warrants review against morphology and matched controls (standard IHC practice). Nuclear only staining needs separate validation because nucleoplasmic localisation is marked uncertain in the subcellular atlas (HPA subcellular). Consider cell identity before calling a region negative: HPA reports high staining in placental cytotrophoblasts and no detected staining in smooth muscle cells (HPA tissue IHC). Persistent color in primary omitted controls implicates detection background, including endogenous enzyme activity, rather than confirmed CCT8 signal (standard IHC practice).
Boster reagents
Best CCT8 / T-complex protein 1 subunit theta IHC Antibodies
PB9993 has IHC images from paraffin sections of mouse kidney, rat testis and human mammary cancer tissue, plus IF data from A549 cells (catalog image captions).
Real IHC data
Anti-TCP1 theta/CCT8 Antibody ®
Cat # PB9993
PB9993 is listed for IHC and IF and reacts with human, mouse and rat samples (catalog applications/reactivity). Its IHC captions show paraffin sections from all three species; its IF caption shows A549 cells (catalog image captions).
Which to pick: Choose PB9993 for tissue IHC when its paraffin-section evidence fits the experiment; the captions report EDTA retrieval at pH 8.0 and 1 μg/ml primary antibody, but do not report the fixative (PB9993 IHC image captions). For IF/ICC, PB9993 is listed for both applications and has an A549 cell IF image (catalog applications; PB9993 IF image caption). It is the only listed cross-species option, with human, mouse and rat reactivity; clonality is unreported (catalog reactivity; catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P50990 (TCPQ_HUMAN, T-complex protein 1 subunit theta).
- Human Protein Atlas. CCT8 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CCT8 subcellular location (ICC-IF): Mainly localized to the nucleoplasm, connecting piece and end piece. In addition localized to the cytosol, mid piece and principal piece..
- Human Protein Atlas. CCT8 antibody validation summary (3 antibodies).
- CCT8 recovers WTp53-suppressed cell cycle evolution and EMT to promote colorectal cancer progression. Oncogenesis 2021 — PMC8642402.
- Multi-Omics Analysis and Verification of the Oncogenic Value of CCT8 in Pan-Cancers. Journal of inflammation research 2023 — PMC10238552.
- CCT8 drives colorectal cancer progression via the RPL4-MDM2-p53 axis and immune modulation. BMC medical genomics 2025 — PMC12008926.
- CCT8 promotes cell migration and tumor metastasis in lung adenocarcinomas. Journal of Cancer 2023 — PMC10622983.
- PubMed PMID:7584026 — UniProt-cited evidence.
- PubMed PMID:8590283 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CCT8 (T-complex protein 1 subunit theta). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9993). Validated IHC antibody: PB9993 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Smooth muscle. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse kidney tissue; fixative not specified (datasheet PB9993); 2. Fixation: Image fixative and duration unreported (datasheet PB9993); 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 PB9993); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9993); 8. Primary antibody: Rabbit anti-CCT8, 0.5-1μg/ml (datasheet PB9993); 9. Primary incubation: Overnight at 4 °C (datasheet PB9993). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9993); Caveat: Smooth muscle cells may be unstained (HPA tissue IHC); Isoform / epitope: 3 isoforms; check epitope coverage for each (UniProt); Regulation: No specific expression regulator annotated (UniProt). Evidence: UniProt P50990, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm, connecting piece and end piece.…, Datasheet PB9993, PMC8642402, PMC10622983. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CCT8/immunohistochemistry-ihc-guide.