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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 using the catalog antibody’s 0.5–1 μg/mL range (datasheet PB9993). Choose controls from HPA’s tissue staining data and assess cytoplasmic staining in the appropriate 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 staining in tissue (HPA tissue IHC) | |
| Staining pattern | General cytoplasmic staining across tissue cells (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 | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Specific expression regulators are not annotated (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended CCT8 IHC & IF Protocols
The catalog antibody’s IHC protocol is followed by published CCT8 protocols for colorectal cancer tissue (PMC8642402) and lung adenocarcinoma tissue microarrays (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 EDTA pH 8.0 heat retrieval (PB9993 datasheet); sodium citrate pH 6.0 is a published alternative for lung tissue microarrays (PMC10622983).
Section 2
What Is the Expected CCT8 Staining Pattern?
CCT8 is a cytoplasmic chaperonin with no transmembrane segment (UniProt P50990). In paraffin section IHC, expect general cytoplasmic staining (HPA: tissue IHC), including high staining in adrenal glandular cells, bronchial respiratory epithelial cells, and placental cytotrophoblasts (HPA: High). HPA rates the tissue pattern Enhanced, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability).
| Distinct cytoplasmic chromogen in adrenal glandular cells, bronchial respiratory epithelium, or placental cytotrophoblasts. | This matches the reported compartment and high-staining cell populations (HPA: general cytoplasmic expression; High in these cells). Judge intensity within the identified cells; neighboring cell types need their own HPA reference. |
| Predominantly nuclear, surface-restricted, or extracellular staining with little cytoplasmic signal. | Recheck localisation before calling this CCT8: tissue IHC is generally cytoplasmic (HPA: tissue IHC), and CCT8 lacks a transmembrane segment (UniProt P50990). Nuclear localisation in ICC-IF is marked uncertain (HPA: subcellular); it does not validate nuclear IHC. |
| Strong staining in smooth muscle cells or chondrocytes while expected positive cells also stain. | These cells were not detected in the HPA tissue survey (HPA: smooth muscle cells; chondrocytes). The discrepancy warrants review for antibody cross-reactivity or endogenous detection activity (general IHC practice); it alone does not identify the cause. |
| Diffuse chromogen over many structures, including spaces between cells, obscures cell boundaries. | This is difficult to score as the reported cytoplasmic pattern (HPA: tissue IHC). Assess no-primary and detection controls for background, and review blocking and washes (general IHC practice) before interpreting intensity. |
| No staining in an otherwise interpretable adrenal glandular cell or bronchial epithelial region. | Both are high-staining populations in the HPA survey (HPA: adrenal gland; bronchus). Check section identity, antibody and detection performance, and antigen retrieval conditions (general IHC practice); an absent signal alone does not establish absent CCT8. |
💡Expected CCT8 appearanceCall a positive result when cytoplasmic staining is clear and high in an HPA high-staining cell population (HPA: tissue IHC); widespread haze or isolated extracellular colour is suspect (general IHC practice).
| Tissue and cell selection | Use an identified high-staining population to judge assay performance (HPA: adrenal glandular cells; bronchial respiratory epithelial cells; placental cytotrophoblasts, High). Smooth muscle cells and chondrocytes are useful low-expectation comparators, not absolute negative controls (HPA: Not detected). |
| Evidence strength | The tissue profile is rated Enhanced, while antibody staining and RNA have medium consistency (HPA: tissue IHC reliability). Three listed antibodies have Enhanced IHC status (HPA: HPA018520, HPA021051, HPA029426). Treat an unexpected pattern as a result to investigate, not as proof of a new localisation. |
| Cellular location and processing | CCT8 is annotated in cytoplasm, centrosome, and cilium basal body; it has no signal peptide or transmembrane segment (UniProt P50990). The expected tissue-level readout remains general cytoplasmic staining (HPA: tissue IHC); the annotations do not promise resolvable centrosomal puncta in chromogenic sections. |
| IF/ICC Q: should nuclear signal be expected? | A: HPA lists nucleoplasm as uncertain and cytosol as supported in ICC-IF, with approved sperm-piece locations (HPA: subcellular). Its IHC pattern is generally cytoplasmic (HPA: tissue IHC). Interpret IF/ICC on its own guide; these observations supply no IF protocol option here. |
| Situation | Likely cause | Next action |
|---|---|---|
| A high-staining reference region is blank. | The result conflicts with the reported high staining, but does not distinguish specimen from assay failure (HPA: adrenal glandular cells; bronchial epithelium, High). | Confirm the cell population and run an appropriate positive section; then review retrieval, primary incubation, and detection controls (general IHC practice). |
| Colour appears mainly in nuclei. | This differs from general cytoplasmic tissue staining (HPA: tissue IHC); nucleoplasm is uncertain in ICC-IF (HPA: subcellular). | Compare a high-staining reference section and a no-primary control; review morphology and detection background before assigning nuclear CCT8 (general IHC practice). |
| Smooth muscle cells or chondrocytes stain strongly. | Both were not detected in the HPA survey (HPA: smooth muscle cells; chondrocytes); cross-reactivity or endogenous detection activity is possible (general IHC practice). | Check a no-primary control and an alternative detection condition, then compare staining with identified high-staining cells (general IHC practice; HPA: tissue IHC). |
| Background makes cytoplasmic borders unreadable. | Diffuse deposit cannot be reliably matched to the reported cytoplasmic profile (HPA: tissue IHC); nonspecific binding or detection background may contribute (general IHC practice). | Review blocking, washes, primary concentration, and no-primary controls; score only cells whose localisation remains interpretable (general IHC practice). |
| Weak staining is seen in oral mucosa or liver cholangiocytes. | Low staining is reported for those populations (HPA: oral mucosa squamous epithelial cells; liver cholangiocytes, Low). | Compare with a high-staining reference population before treating a weak signal as assay failure (HPA: adrenal glandular cells, High; general IHC practice). |
| Puncta near nuclei are the only apparent signal. | Centrosome localisation is annotated (UniProt P50990), but the tissue survey describes general cytoplasmic expression (HPA: tissue IHC). | Review a high-staining tissue region for cytoplasmic signal and check no-primary background; do not score puncta alone as a validated chromogenic pattern (general IHC practice; HPA: tissue IHC). |
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 bronchus first: respiratory epithelial cells should stain strongly (HPA: High in bronchus respiratory epithelial cells). Use smooth muscle as the negative tissue; bronchial smooth muscle cells on the positive slide should show only background staining (HPA: smooth muscle cells Not detected).
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, a rabbit isotype control matched to the catalog antibody’s clonality, and CCT8 knockout material as a biological negative (selected PB9993 tissue-IHC caption: rabbit primary antibody). For chromogenic detection, quench endogenous peroxidase and block endogenous biotin when using the caption’s biotinylated secondary and streptavidin–biotin/DAB method (selected PB9993 tissue-IHC caption).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected PB9993 tissue-IHC caption does not state the fixative (selected PB9993 tissue-IHC caption). Paraffin-section IHC has a documented starting condition of heat retrieval in EDTA at pH 8.0; the evidence does not establish whether retrieval is required under other conditions (selected PB9993 tissue-IHC caption). Frozen sections and IF have no matched protocol in the supplied evidence, so their relative ease is unknown; bronchial smooth muscle provides a local background check (HPA: smooth muscle cells Not detected).
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 staining in paraffin sections by checking retrieval, cellular location, controls and scoring before interpreting chromogenic signal.
Which retrieval conditions should I start with for CCT8 in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet PB9993). That condition accompanies CCT8 detection in mouse kidney with 1 μg/ml primary antibody overnight at 4°C (PB9993 tissue-IHC caption). If cytoplasmic staining is weak, compare a small retrieval time series while keeping section thickness, antibody concentration and detection constant (standard IHC practice). Include a matched section with the primary antibody omitted to identify detection-system background (standard IHC practice). Evaluate whether the change improves cellular signal without increasing tissue damage or diffuse chromogen deposition (standard IHC practice).
Could fixation explain weak or uneven CCT8 staining?
The selected paraffin-section caption does not report a fixative, so target-specific fixation sensitivity is unknown (PB9993 tissue-IHC caption). Record the actual fixative and fixation duration for each specimen before comparing CCT8 intensity across blocks (standard IHC practice). For weak staining, compare sections processed with documented fixation histories under the same EDTA pH 8.0 retrieval and primary-antibody conditions (datasheet PB9993; standard IHC practice). Check tissue preservation and whether staining varies with section depth or specimen edges before changing antibody concentration (standard IHC practice). Neither tissue staining patterns nor CCT8 topology establishes a fixation effect for this antibody (HPA tissue IHC; UniProt P50990).
Where should CCT8 staining appear, and how should I handle nuclear signal?
Expect broadly cytoplasmic tissue staining as the starting pattern for chromogenic interpretation (HPA tissue IHC: general cytoplasmic expression). CCT8 is annotated in cytoplasm, centrosome and cilium basal body, with no transmembrane segment (UniProt P50990). HPA subcellular imaging also reports cytosol and an uncertain nucleoplasmic assignment, so isolated nuclear DAB staining needs independent corroboration (HPA subcellular). Compare cellular staining with the primary-omission control and assess whether chromogen follows intact cell boundaries rather than section edges (standard IHC practice). Document the compartment scored and use that same compartment across specimens; do not combine nuclear and cytoplasmic scores without a stated rule (standard IHC practice).
Can this antibody distinguish CCT8 isoforms or modified epitopes in tissue?
CCT8 has 3 annotated isoforms, but the supplied product caption does not identify the antibody epitope (UniProt P50990; PB9993 tissue-IHC caption). Therefore, a positive chromogenic stain cannot be assigned to a particular isoform from this evidence alone (UniProt P50990; PB9993 tissue-IHC caption). Annotated modifications include phosphorylation at Ser23 and Ser162 and acetylation at Ala2; their effects on tissue staining are unreported here (UniProt P50990). If isoform specificity matters, first obtain the immunogen or epitope sequence and compare it with the isoform sequences (standard antibody-validation practice). Report the result as CCT8 immunoreactivity until that comparison and an independent specificity check support a narrower claim (standard antibody-validation practice).
How should I assess CCT8 by IF alongside a cell-type marker?
For the separate IF/ICC application, multiplex CCT8 with a validated cytotrophoblast marker when examining placenta, where cytotrophoblast staining is reported as high (HPA tissue IHC). Choose spectrally separated fluorophores and consider a far-red channel for the weaker signal when tissue autofluorescence affects shorter wavelengths (standard IF practice). CCT8 has no transmembrane segment and is annotated in the cytoplasm, so assess permeabilisation for access to an intracellular epitope (UniProt P50990; standard IF practice). Run single-stain and secondary-only controls to identify bleed-through and nonspecific fluorescence (standard IF practice). Compare the IF compartment with the reported cytosolic signal while treating the nucleoplasmic assignment as uncertain (HPA subcellular).
How can I separate CCT8 staining from chromogenic background?
The selected tissue demonstration used 10% goat serum, a biotinylated secondary antibody and DAB development (PB9993 tissue-IHC caption). If diffuse brown signal persists, inspect a primary-omission section and review blocking, washes and DAB development time under otherwise matched conditions (standard IHC practice). Apply a peroxidase block as part of the general chromogenic workflow, then assess whether residual signal tracks tissue structures or the intended cellular compartment (standard IHC practice). Because the demonstrated detection uses biotin, evaluate endogenous biotin as a possible source of background where appropriate (PB9993 tissue-IHC caption; standard IHC practice). Recheck primary concentration against the demonstrated 1 μg/ml before increasing it (PB9993 tissue-IHC caption).
What is a defensible way to quantify CCT8 chromogenic staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cellular compartment as cytoplasm before analysis, consistent with the general tissue pattern (HPA tissue IHC). Within each annotated cell population, report the percentage of positive cells and an intensity-weighted H-score of 0–300, using fixed intensity thresholds (standard IHC scoring practice). If counting positive cells per mm², normalise to evaluable tissue area and exclude folds, necrosis and section edges by a prespecified rule (standard IHC practice). Keep retrieval, antibody concentration, chromogen development and imaging settings consistent across the comparison (standard IHC practice). Report cell populations separately because HPA describes high cytotrophoblast staining and undetected smooth-muscle-cell staining (HPA tissue IHC).
When should I question a strong CCT8-positive result?
A strong cytoplasmic signal in intact cells fits the reported general tissue pattern, while staining confined to an unexpected compartment warrants verification (HPA tissue IHC; standard IHC practice). Check cell identity: cytotrophoblasts are reported high, whereas smooth muscle cells are reported undetected in the HPA tissue survey (HPA tissue IHC). Treat signal limited to section edges, folds or necrotic regions as a possible processing artefact and compare it with well-preserved tissue (standard IHC practice). Inspect the primary-omission control for endogenous enzyme or detection-system signal before calling a section positive (standard IHC practice). HPA rates the tissue staining evidence Enhanced but describes only medium consistency with RNA expression, so report discordance explicitly (HPA tissue IHC).
Boster reagents
Best CCT8 / T-complex protein 1 subunit theta IHC Antibodies
PB9993 has real IHC images from paraffin-embedded mouse kidney, rat testis and human mammary cancer tissue, plus an IF image from A549 cells (PB9993 image captions).
Real IHC data
Anti-TCP1 theta/CCT8 Antibody ®
Cat # PB9993
PB9993 is listed for IHC, IHC-F, IF and ICC in human, mouse and rat (PB9993 catalog applications and reactivity). Its images show paraffin-section IHC in mouse kidney, rat testis and human mammary cancer, and IF in A549 cells (PB9993 image captions).
Which to pick: For tissue IHC, choose PB9993: its IHC captions document paraffin-embedded sections, 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 (PB9993 catalog applications; PB9993 IF image caption). For cross-species work, PB9993 lists human, mouse and rat reactivity; its clonality is unreported (PB9993 catalog reactivity and 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 staining in tissue (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: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unknown (UniProt); Regulation: Specific expression regulators are not 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.