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CCT6A / T-complex protein 1 subunit zeta · IHC design guide
Design Immunohistochemistry for CCT6A
Plan CCT6A chromogenic IHC around the reported cytoplasmic tissue pattern (HPA tissue IHC). Compare strong staining in colon glandular cells with undetected staining in ovarian stroma, while accounting for possible detection of products from more than one gene (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CCT6A 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 | General cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic signal in glandular, epithelial and neuronal cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Ovary |
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 | Staining may reflect products of more than one gene (HPA tissue IHC) | |
| Regulation | Low tissue specificity; regulation unreported (HPA tissue IHC) | |
| Isoform / epitope | Two isoforms; epitope differences are unreported (UniProt) |
Section 1
Recommended CCT6A IHC & IF Protocols
The catalog antibody’s IHC-P protocol appears alongside four published CCT6A IHC workflows (PMC8418478; PMC11574388; PMC12174516; PMC7850773).
| Sample | Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A09373-1) |
| Fixation | Image fixative and duration unreported (datasheet A09373-1); 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 citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane 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-CCT6A, 1:100-1:300 (datasheet A09373-1) |
| 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 | CCT6A-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 citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule); compare microwave EDTA retrieval for astrocytoma FFPE sections (PMC8418478).
Section 2
What Is the Expected CCT6A Staining Pattern?
CCT6A is a cytoplasmic protein with no transmembrane segment (UniProt P40227). In paraffin section IHC, expect cytoplasmic staining in several epithelial, glandular and neuronal cell populations reported as High by HPA (HPA: tissue IHC). HPA describes general cytoplasmic expression, but rates its tissue pattern Supported with medium consistency and cautions that the antibodies may target proteins from more than one gene (HPA: tissue IHC).
| Cytoplasmic chromogen in colon or duodenal glandular cells, bronchial respiratory epithelium, or cortical neurons (HPA: High in these cell types). | The compartment and cell types fit the reference pattern (UniProt P40227: cytoplasm; HPA: tissue IHC). Compare intensity within the same run; an HPA High designation is a reference observation, not a required score for every specimen (general IHC practice). |
| Predominantly nuclear or crisp membrane staining, with little cytoplasmic signal (UniProt P40227: cytoplasm; HPA: general cytoplasmic expression). | Treat this as a discordant pattern requiring an artefact check (general IHC practice). UniProt reports no transmembrane segment (UniProt P40227); location alone cannot identify the source of an unexpected signal (general IHC practice). |
| Strong staining in ovarian stroma or smooth muscle, especially when expected epithelial or glandular cells are faint (HPA: ovarian stroma Not detected; smooth muscle Low). | Check for nonspecific antibody binding or endogenous detection activity (general IHC practice). HPA cautions that its antibodies may target proteins from more than one gene, so staining in an unexpected cell type cannot establish CCT6A expression by itself (HPA: tissue IHC). |
| Diffuse chromogen over cells and surrounding tissue, without a discernible cytoplasmic pattern (HPA: general cytoplasmic expression). | A broadly distributed deposit is difficult to assign to CCT6A (general IHC practice). Review negative controls, washing and detection background before scoring cells; HPA's Supported rating does not validate a particular slide's background (HPA: tissue IHC; general IHC practice). |
| No staining in a run that includes colon glandular cells or bronchial respiratory epithelium (HPA: High in these cell types). | First investigate assay performance with controls and the catalog antibody's IHC-P instructions (general IHC practice). A blank section does not establish absence of CCT6A when a reference positive cell population also fails (HPA: tissue IHC; general IHC practice). |
💡Expected CCT6A appearanceCall a result consistent with the reference when chromogen is chiefly cytoplasmic in HPA High epithelial, glandular or neuronal cells (UniProt P40227: cytoplasm; HPA: tissue IHC); prominent nuclear, membrane or indiscriminate stromal staining warrants review as a possible false positive (HPA: tissue IHC; general IHC practice).
| Cellular compartment (UniProt P40227; HPA: tissue IHC) | UniProt assigns CCT6A to the cytoplasm and reports no transmembrane segment (UniProt P40227). HPA describes general cytoplasmic expression in tissue (HPA: tissue IHC). Use those observations to assess localisation before comparing staining intensity (general IHC practice). |
| Reference cell populations (HPA: tissue IHC) | HPA reports High staining in adrenal and intestinal glandular cells, respiratory epithelium, esophageal squamous epithelium and neurons in caudate and cerebral cortex (HPA: tissue IHC). It reports Low smooth muscle staining and no detected ovarian stromal staining (HPA: tissue IHC). |
| Strength of tissue evidence (HPA: tissue IHC) | The tissue profile is Supported, with medium consistency between antibody staining and RNA expression, and external verification pending (HPA: tissue IHC). HPA also warns that the antibodies target proteins from more than one gene (HPA: tissue IHC); interpret unexpected positives cautiously. |
| Antibody evidence (HPA: antibody validation) | HPA lists IHC as Supported for HPA042996, HPA045576 and HPA049949; it lists ICC as Supported for HPA049949 (HPA: antibody validation). These statuses describe those antibodies, so check which antibody produced the image being compared (general IHC practice). |
| IF/ICC Q: Where should signal appear? (HPA: subcellular ICC-IF) | A: HPA's approved main location is the cytosol, with images listed for A-431, PC-3 and U2OS (HPA: subcellular ICC-IF). HPA cautions that this localisation used antibodies targeting proteins from multiple genes (HPA: subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected cytoplasmic signal is absent in an HPA High cell population (HPA: tissue IHC). | The run may have failed at staining or detection; one blank specimen cannot distinguish assay failure from biology (general IHC practice). | Check a reference positive section and detection controls, then review retrieval, antibody dilution and incubation against the catalog antibody's IHC-P instructions (general IHC practice). |
| Nuclei or cell borders dominate while cytoplasm is weak (UniProt P40227: cytoplasm; HPA: tissue IHC). | The compartment differs from the supplied localisation evidence; background or nonspecific binding is possible (general IHC practice). | Inspect the negative control and compare the same cell types across sections before assigning a positive score (general IHC practice). |
| Chromogen is widespread outside identifiable cells (HPA: general cytoplasmic expression). | Diffuse background can obscure whether cytoplasmic staining is present (general IHC practice). | Review blocking, washes and the detection control; score only cell-associated staining that can be distinguished from background (general IHC practice). |
| Ovarian stromal cells stain strongly (HPA: Not detected in ovarian stroma). | The result conflicts with HPA's reference observation; HPA warns of antibodies targeting more than one gene (HPA: tissue IHC). | Check endogenous detection activity and an appropriate negative control, then seek independent confirmation before attributing the signal to CCT6A (general IHC practice). |
| A tissue stains more weakly or unevenly than an HPA High example (HPA: tissue IHC). | The HPA level describes its observed samples; differences in specimen or assay conditions can affect comparison between IHC runs (HPA: tissue IHC; general IHC practice). | Compare an internal or same-run reference cell population and document compartment, cell type and intensity separately (general IHC practice). |
Experiment evidence
IHC & IF images showing CCT6A staining
A09373-1 · IHCImmunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue, using CCT6A Antibody. The picture on the right is blocked
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000146731-CCT6A/tissue/cerebral+cortex#imid_12484037
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000146731-CCT6A/tissue/colon#imid_12483979
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000146731-CCT6A/tissue/liver#imid_12483964
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000146731-CCT6A/tissue/kidney#imid_12483981
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA049949; green = CCT6A, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA049949; green = CCT6A, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CCT6A IHC & IF
🧪Run colon first: glandular cells must show strong staining (HPA: High in colon glandular cells). Run ovary as the negative tissue, focusing on ovarian stroma cells (HPA: Not detected); on the colon slide, use adjacent nonglandular cells as an internal background comparator and score them as negative only if they lack specific cytoplasmic staining (UniProt P40227: cytoplasm).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Ovary (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CCT6A in A-431, PC-3, U2OS, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species and immunoglobulin class (standard IHC practice). Use peptide preabsorption as a target-specific control (selected-SKU tissue-IHC caption: staining blocked with synthesized peptide), and quench endogenous peroxidase before chromogenic detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU paraffin-section caption does not state a fixative (selected-SKU tissue-IHC caption: fixative not stated). Retrieval dependency is unreported; assess antigen retrieval empirically for paraffin sections (standard IHC practice). There is no supplied evidence that frozen sections or IF are easier; in colon, distinguish staining of glandular-cell cytoplasm from signal in luminal material when scoring (HPA: High in colon glandular cells; UniProt P40227: cytoplasm).
HPA tissue IHC evidence for CCT6A
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene. 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CCT6A IHC Tips
Troubleshoot CCT6A staining in paraffin sections by checking retrieval, cytoplasmic localisation, controls and scoring before interpreting chromogenic signal.
What retrieval conditions should I try when CCT6A staining is weak in paraffin sections?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval setting). Allow sections to cool in the retrieval buffer, then compare staining with a matched section processed in the same run (standard IHC practice). CCT6A is cytoplasmic and has no transmembrane segment, so evaluate recovered signal in cytoplasm rather than treating nuclear colour as successful retrieval (UniProt P40227 localisation and topology). If staining remains weak, optimise retrieval time on adjacent sections while holding antibody concentration and detection conditions constant (standard IHC practice). Compare intact tissue regions and inspect morphology, because harsh retrieval can make stronger colour harder to interpret (standard IHC practice).
How should I troubleshoot uneven CCT6A staining when fixation conditions are uncertain?
The selected image documents paraffin-embedded human breast carcinoma and peptide-blocked staining, but does not report its fixative (caption A09373-1). CCT6A-specific sensitivity to fixation is therefore unknown; neither tissue staining patterns nor the protein’s cytoplasmic location establish a preferred fixation condition (caption A09373-1; UniProt P40227 localisation). Record the fixative, fixation duration and processing history for each specimen before comparing staining intensity (standard IHC practice). On available sections, compare well-preserved central tissue with edges and damaged areas, and keep retrieval and detection conditions identical across comparisons (standard IHC practice). Interpret differences cautiously when fixation histories differ, even if both sections show cytoplasmic colour (standard IHC practice; UniProt P40227 localisation).
Which staining compartment supports a CCT6A-positive IHC result?
Expect predominantly cytoplasmic staining: UniProt places CCT6A in the cytoplasm, and HPA describes general cytoplasmic tissue expression (UniProt P40227 localisation; HPA tissue IHC). HPA also reports an approved cytosolic ICC/IF location, although that annotation cautions that its antibodies target proteins from multiple genes (HPA subcellular). Score cytoplasmic signal in morphologically intact cells and review isolated nuclear or surface staining as possible artefact before calling it positive (standard IHC practice; UniProt P40227 localisation). The protein has no transmembrane segment, so a crisp membrane-only pattern needs independent validation (UniProt P40227 topology). Compare the questioned pattern with a matched negative control and the peptide-blocked image where relevant (standard IHC practice; caption A09373-1).
Could isoforms or epitope accessibility explain discordant CCT6A IHC staining?
UniProt lists 2 CCT6A isoforms, but the supplied evidence does not map the catalog antibody’s epitope to either one (UniProt P40227 isoforms; caption A09373-1). Consequently, do not interpret a staining difference between specimens as an isoform switch from this IHC result alone (standard IHC interpretation). CCT6A has annotated acetylated and phosphorylated residues, yet the payload does not establish whether those modifications affect antibody binding in sections (UniProt P40227 modified residues). Compare adjacent sections under identical retrieval and detection conditions, and document the antibody’s stated immunogen or epitope if that information becomes available (standard IHC practice). The peptide-blocked breast carcinoma image supports competition for that reagent’s staining, without identifying the isoform or modified state detected (caption A09373-1).
How can IF help check cell identity and localisation alongside CCT6A IHC?
Use IF/ICC as a separate localisation check: HPA reports approved cytosolic staining but cautions that its antibodies may recognise proteins from multiple genes (HPA subcellular). For a respiratory epithelial specimen, multiplex a cell identity marker such as pan-cytokeratin and assess whether cytosolic CCT6A signal lies within the marked cells (HPA tissue IHC: respiratory epithelial cells High; standard IF practice). Choose a fluorophore channel after measuring tissue autofluorescence, favouring a spectrally distinct channel when background is strong (standard IF practice). Because CCT6A is cytoplasmic and lacks a transmembrane segment, permeabilisation should allow access to an intracellular epitope; optimise it for the IF specimen (UniProt P40227 localisation and topology; standard IF practice). Do not transfer the paraffin image’s unreported fixation conditions to IF (caption A09373-1).
What controls help separate diffuse CCT6A signal from chromogenic background?
CCT6A has general cytoplasmic expression, so broad cytoplasmic colour alone is insufficient to establish specificity (HPA tissue IHC; UniProt P40227 localisation). Include a section without primary antibody to assess detection background, and use a peroxidase block before chromogenic development when using a peroxidase-based system (standard IHC practice). Check pigment, tissue folds, necrosis and section edges before scoring diffuse or granular signal (standard IHC practice). The selected breast carcinoma image includes a synthesized-peptide blocking comparison for SKU A09373-1, which can inform interpretation of that reagent’s staining (caption A09373-1). HPA’s tissue profile is marked Supported but notes that its antibodies target proteins from more than one gene, so corroborate unexpected cell patterns independently (HPA tissue IHC).
How should I quantify CCT6A across heterogeneous paraffin specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and intact tissue area before scoring cytoplasmic CCT6A staining (UniProt P40227 localisation; standard IHC practice). An H-score combines the percentage of cells at each intensity with intensity grades, while percentage-positive scoring records the stained fraction using a prespecified threshold (standard IHC practice). If spatial distribution matters, count positive cells per mm² of evaluable tissue and report the area assessed (standard IHC practice). Normalise comparisons to the same cell type, tissue compartment and staining run, excluding folds, necrosis and poorly preserved regions (standard IHC practice). HPA reports low tissue RNA specificity and varied cell-level staining, so pooling unlike cell populations can obscure a meaningful difference (HPA tissue IHC).
When is a CCT6A-positive IHC pattern credible rather than artefactual?
A credible pattern places signal mainly in intact cell cytoplasm, consistent with UniProt localisation and HPA’s general cytoplasmic tissue profile (UniProt P40227 localisation; HPA tissue IHC). Check cell identity against morphology: HPA reports high staining in several epithelial and neuronal populations, whereas ovarian stroma is listed as not detected (HPA tissue IHC). Review nuclear-only or membrane-only colour, edge enhancement and necrotic regions before calling them true CCT6A signal (UniProt P40227 localisation and topology; standard IHC practice). A no-primary control helps identify detection background, including endogenous enzyme activity in peroxidase-based staining (standard IHC practice). Peptide competition in the selected breast carcinoma image supports interpretation of that reagent’s staining, but does not resolve HPA’s cross-gene antibody caution (caption A09373-1; HPA tissue IHC).
Boster reagents
Best CCT6A / T-complex protein 1 subunit zeta IHC Antibodies
Catalog anti-CCT6A antibodies list IHC reactivity with human, mouse, and rat (catalog: applications and reactivity); one has a human paraffin-section IHC image (A09373-1 image caption), while IF is catalog-listed (A09373-1 applications).
Real IHC data
Anti-TCP-1 zeta CCT6A Antibody
Cat # A09373-1
A09373-1 lists IHC and IF for human, mouse, and rat (catalog: applications and reactivity); its IHC image shows a human breast carcinoma paraffin section with a peptide-blocked comparison (A09373-1 image caption). A09373-2 lists IHC for human, mouse, and rat (catalog: applications and reactivity), with no IHC or IF image supplied (catalog: image captions).
Which to pick: For tissue IHC, choose A09373-1 when a matching paraffin-section example matters (A09373-1 image caption); the caption does not report the fixative (A09373-1 image caption). For IF, choose A09373-1 because IF is listed for that SKU (catalog: applications); neither SKU supplies an IF image or an ICC application claim (catalog: applications and image captions). For mouse or rat samples, both SKUs list reactivity (catalog: reactivity), but the supplied IHC image documents only a human sample (A09373-1 image caption; A09373-2 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P40227 (TCPZ_HUMAN, T-complex protein 1 subunit zeta).
- Human Protein Atlas. CCT6A tissue IHC expression (reliability: Supported).
- Human Protein Atlas. CCT6A subcellular location (ICC-IF): Localized to the cytosol. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. CCT6A antibody validation summary (3 antibodies).
- Chaperonin-containing tailless complex polypeptide 1 subunit 6A correlates with increased World Health Organization grade, less isocitrate dehydrogenase mutation, and deteriorative survival of astrocytoma patients. Journal of clinical laboratory analysis 2021 — PMC8418478.
- CCT6A alleviates pulmonary fibrosis by inhibiting HIF-1α-mediated lactate production. Journal of molecular cell biology 2024 — PMC11574388.
- Prognostic Significance of Chaperonin-Containing Tailless Complex Polypeptide 6A (CCT6A) in Ewing Sarcoma. Asian Pacific journal of cancer prevention : APJCP 2025 — PMC12174516.
- CCT6A, a novel prognostic biomarker for Ewing sarcoma. Medicine 2021 — PMC7850773.
- PubMed PMID:8034610 — UniProt-cited evidence.
- PubMed PMID:12853948 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CCT6A (T-complex protein 1 subunit zeta). Expected localisation General cytoplasmic staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A09373-1 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Ovary. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A09373-1); 2. Fixation: Image fixative and duration unreported (datasheet A09373-1); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 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-CCT6A, 1:100-1:300 (datasheet A09373-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Staining may reflect products of more than one gene (HPA tissue IHC); Isoform / epitope: Two isoforms; epitope differences are unreported (UniProt); Regulation: Low tissue specificity; regulation unreported (HPA tissue IHC). Evidence: UniProt P40227, HPA tissue IHC · Supported, HPA ICC-IF · cytosol. Caution: Based on antibodies targeting proteins from…, Datasheet A09373-1, PMC8418478, PMC11574388. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CCT6A/immunohistochemistry-ihc-guide.