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CCT5 / T-complex protein 1 subunit epsilon · IHC design guide
Design Immunohistochemistry for CCT5
Plan CCT5 paraffin-section IHC around a cytoplasmic pattern, using seminiferous duct cells as a strong tissue reference (HPA tissue IHC). The catalog antibody's IHC starting range is 0.5–1 μg/mL (datasheet PB9928).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CCT5 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 | Cytoplasmic; most abundant in seminiferous duct cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet PB9928) | |
| Positive control | Caudate+4 more · see all | |
| Negative control | Esophagus+4 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 PB9928) | |
| Caveat | CNS nuclear positivity is considered off target (HPA tissue IHC) | |
| Regulation | Stimulus-dependent regulation is not annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope impact is undetermined (UniProt) |
Section 1
Recommended CCT5 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: PB9928) with 3 published CCT5 IHC protocols (PMC5609992; PMC11409311; PMC13014677).
| Sample | Paraffin-embedded human intestinal cancer tissues; fixative not specified (datasheet PB9928) |
| Fixation | Image fixative and duration unreported (datasheet PB9928); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet PB9928) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9928) |
| Primary antibody | Rabbit anti-CCT5, 0.5-1μg/ml (datasheet PB9928) |
| Primary incubation | Overnight at 4 °C (datasheet PB9928) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9928) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CCT5-positive staining in glial cells of caudate (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression, most abundant in seminiferous duct cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate pH 6 retrieval (datasheet: PB9928); a published protocol also uses heated citrate, but does not report its pH (PMC11409311).
Section 2
What Is the Expected CCT5 Staining Pattern?
In paraffin IHC, expect predominantly cytoplasmic CCT5 staining across many tissues, with the strongest reported staining in seminiferous duct cells, including elongated or late spermatids (HPA: Enhanced tissue IHC; general cytoplasmic expression; High in elongated or late spermatids). CCT5 is a cytoplasmic protein also annotated at the centrosome and has no transmembrane segment (UniProt P48643: subcellular location; topology). Interpret intensity by cell type rather than expecting every cell in a section to stain equally (HPA: cell-specific tissue levels).
| Clear cytoplasmic staining in late spermatids; cytoplasmic staining in other reported High populations. | This fits the reported pattern: testis elongated or late spermatids, caudate glia, lymph-node germinal-center cells, placental cytotrophoblasts, and fallopian-tube or parathyroid glandular cells are High (HPA: tissue IHC). Compare the correct cell population and compartment, not whole-section darkness (HPA: general cytoplasmic expression; cell-specific levels). |
| Prominent nuclear staining, particularly in CNS cells, with little convincing cytoplasmic signal. | Treat this as suspect: HPA explicitly disregards CNS nuclear positivity as off target, while its tissue profile is cytoplasmic (HPA: reliability description; tissue profile). UniProt places CCT5 in cytoplasm and at the centrosome, not the nucleus (UniProt P48643: subcellular location). |
| Strong staining in a population HPA lists as Not detected, such as cardiomyocytes or esophageal squamous epithelium. | This conflicts with the reported cell-specific reference pattern and warrants checks for cross-reactivity or endogenous detection activity (HPA: Not detected in cardiomyocytes and esophageal squamous epithelial cells; general IHC practice). A negative HPA category applies to the named cell population, not necessarily every cell in that tissue (HPA: tissue IHC). |
| Diffuse color across cells and extracellular space, without discernible cytoplasmic boundaries. | This does not reproduce HPA's cell-associated cytoplasmic pattern (HPA: tissue IHC profile). Review primary-omission and detection controls, washing, and blocking as general IHC background checks; the supplied records do not identify a CCT5-specific background mechanism (general IHC practice). |
| No signal in elongated or late spermatids, while the section and detection controls appear adequate. | This misses a reported High population and calls for review of tissue identity, the antibody's IHC-P instructions, and control performance (HPA: High in elongated or late spermatids; general IHC practice). HPA's Enhanced reliability supports use as a reference pattern but does not guarantee staining in every preparation (HPA: Enhanced tissue IHC). |
💡Expected CCT5 appearanceA convincing positive shows cell-associated cytoplasmic staining, especially strong in testis elongated or late spermatids, whereas dominant CNS nuclear color is suspect (HPA: general cytoplasmic expression; High in late spermatids; CNS nuclear positivity disregarded as off target).
| Compartment and topology (UniProt P48643: subcellular location; topology). | CCT5 is cytoplasmic, with a centrosomal annotation and no transmembrane segment; assess chromogenic IHC chiefly for a cytoplasmic pattern (UniProt P48643; HPA: general cytoplasmic expression). Centrosomal annotation alone does not establish a reliably resolvable punctum in paraffin sections (UniProt P48643; general IHC practice). |
| Cell-specific reference intensity (HPA: tissue IHC). | High and Not detected labels refer to specified cells within tissues; for example, placental cytotrophoblasts are High and liver cholangiocytes are Not detected (HPA: tissue IHC). HPA also reports low tissue RNA specificity, so a named negative cell population is a comparison control, not a claim that the entire organ lacks CCT5 (HPA: RNA specificity; tissue IHC). |
| Antibody validation (HPA: antibody records; tissue IHC). | Two listed rabbit polyclonal antibodies have Enhanced IHC status; the listed mouse monoclonal has Supported IHC status (HPA: HPA002238, HPA005958, CAB006271). HPA reports high agreement between staining and RNA data while expressly excluding CNS nuclear positivity from the accepted pattern (HPA: reliability description). |
| Isoforms and processing (UniProt P48643: isoforms; processing). | UniProt lists two isoforms, no signal peptide or propeptide, and a chain spanning residues 2–541 (UniProt P48643). The supplied records do not map the catalog antibody's epitope or establish isoform coverage; do not infer either from a positive IHC result (UniProt P48643; HPA: antibody records). |
| Target-specific fixation sensitivity (HPA: tissue IHC; UniProt P48643). | Neither supplied source reports a CCT5-specific fixation effect. If signal varies between preparations, investigate preparation and detection variables as general IHC practice without attributing the difference to documented CCT5 epitope masking (general IHC practice; HPA: tissue IHC; UniProt P48643). |
| Situation | Likely cause | Next action |
|---|---|---|
| Strong nuclear color in a CNS section. | The pattern matches the nuclear positivity HPA disregards as off target (HPA: reliability description). | Score cytoplasmic staining separately; compare a reported High cell population and a primary-omission control before accepting the result (HPA: tissue IHC; general IHC practice). |
| Color appears in a reported Not detected cell population. | Cross-reactivity or endogenous detection activity is possible; the HPA category is cell-specific (HPA: tissue IHC; general IHC practice). | Confirm the cell identity, review primary-omission and detection controls, and compare the same run with a reported High population (HPA: tissue IHC; general IHC practice). |
| The entire section has diffuse brown background. | Nonspecific binding, inadequate blocking or washing, or endogenous detection activity are general IHC possibilities (general IHC practice). | Inspect controls and the detection system; optimize blocking and washes according to the IHC-P workflow without assuming a CCT5-specific cause (general IHC practice). |
| Reported High late spermatids show no convincing cytoplasmic signal. | The result conflicts with the HPA reference pattern; tissue selection or assay performance needs checking (HPA: High in late spermatids; general IHC practice). | Verify the cell population, follow the catalog antibody's IHC-P instructions, and check positive and detection controls. No target-specific fixation sensitivity is documented here (HPA: tissue IHC; general IHC practice). |
| Staining is faint in bronchial respiratory epithelium or kidney glomerular cells. | HPA reports Low staining in these named populations, so faint signal may match the reference (HPA: tissue IHC). | Judge them against their Low category and compare a reported High population in the same run before changing the assay (HPA: tissue IHC; general IHC practice). |
| IF/ICC Q&A: Does sperm mid-piece fluorescence validate a paraffin IHC pattern? | HPA approves sperm mid-piece localization, with additional principal-piece and annulus localization, in ICC-IF images (HPA: subcellular ICC-IF). | Use it only as an IF localization reference; judge paraffin IHC against HPA's tissue cytoplasmic and cell-specific patterns (HPA: subcellular ICC-IF; tissue IHC). |
Experiment evidence
IHC & IF images showing CCT5 staining
PB9928 · IHCIHC analysis of CCT5 using anti-CCT5 antibody (PB9928). CCT5 was detected in paraffin-embedded section of human intestinal cancer tissues. H
PB9928 · IHCIHC analysis of CCT5 using anti-CCT5 antibody (PB9928). CCT5 was detected in paraffin-embedded section of mouse ovary tissues. Heat mediated
PB9928 · IFIF analysis of CCT5 using anti-CCT5 antibody (PB9928). CCT5 was detected in immunocytochemical section of U20S cell. Enzyme antigen retrieva
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000150753-CCT5/tissue/cerebral+cortex#imid_2048232
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000150753-CCT5/tissue/colon#imid_2048166
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000150753-CCT5/tissue/liver#imid_2063935
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000150753-CCT5/tissue/kidney#imid_2063944
HPA · ICC-IF SpermHuman Protein Atlas immunofluorescence, Sperm, antibody HPA002238; green = CCT5, blue = nucleus, red = microtubules.
HPA · ICC-IF SpermHuman Protein Atlas immunofluorescence, Sperm, antibody HPA002238; green = CCT5, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CCT5 IHC & IF
🧪Run lymph node first and look for staining in germinal center cells (HPA: High in lymph node germinal center cells). Use heart muscle cardiomyocytes as a negative tissue comparator (HPA: Not detected in cardiomyocytes); on the lymph node slide, compare germinal centers with cells that remain unstained, without assuming a named cell population is CCT5-negative.
Positive control tissue: Caudate (Glial cells, HPA High)
Negative control tissue: Esophagus (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CCT5 in Sperm, with annotated localisation: Mid piece (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and nonimmune rabbit IgG matched to the catalog primary antibody (selected-SKU caption: rabbit anti-CCT5); use CCT5 knockout material as a biological negative if available. In lymph node, check for endogenous peroxidase background before interpreting chromogenic signal (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption: fixative unreported). The caption reports heat retrieval in citrate buffer at pH 6 for 20 minutes, but does not establish whether CCT5 staining depends on retrieval (selected-SKU caption: citrate retrieval). No matched frozen-section or IF comparison establishes an easier option; for lymph node, distinguish specific germinal center staining from peroxidase background (HPA: High in germinal center cells; standard IHC practice).
HPA tissue IHC evidence for CCT5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Nuclear positivity in CNS viewed as off target and disregarded.). 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 |
|---|---|---|---|---|
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
| Placenta | Cytotrophoblasts | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Vagina | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CCT5 IHC Tips
Troubleshoot CCT5 staining in paraffin sections by checking retrieval, cytoplasmic localisation, detection controls and cell-specific scoring.
How should I adjust retrieval when CCT5 staining is weak in paraffin sections?
Start CCT5 IHC with heat-mediated citrate retrieval at pH 6 for 20 minutes (datasheet PB9928; caption PB9928). Keep the caption’s 1 µg/mL primary incubation overnight at 4°C while comparing adjacent sections (caption PB9928). If signal remains weak, vary retrieval duration while holding detection constant, and watch for section loss or edge enhancement (standard IHC practice). Document heating uniformity and cooling, since uneven retrieval can resemble heterogeneous expression (standard IHC practice). Score recovered cytoplasmic signal; CCT5 is cytoplasmic and has no transmembrane segment (UniProt P48643).
Can fixation explain variable CCT5 staining between paraffin blocks?
The PB9928 image identifies paraffin-embedded human intestinal cancer tissue but does not state its fixative, so CCT5-specific fixation sensitivity is unknown (caption PB9928). Record each block’s fixative, exposure time and processing history before attributing intensity differences to CCT5 expression (standard IHC practice). For a controlled comparison, start with citrate at pH 6 for 20 minutes and 1 µg/mL primary antibody overnight at 4°C (caption PB9928). Run a reference section alongside the samples, compare preserved regions, and exclude folds, crush and poorly preserved areas from scoring (standard IHC practice).
What CCT5 staining pattern should I expect, and when is nuclear staining suspect?
Expect predominantly cytoplasmic CCT5 staining in chromogenic tissue IHC rather than a nuclear or cell-surface pattern (HPA tissue IHC; UniProt P48643). CCT5 is also annotated at the cytoskeleton and centrosome, although routine DAB sections may not resolve those structures; interpret fine puncta cautiously (UniProt P48643; standard IHC practice). HPA reports its greatest general abundance in seminiferous duct cells and high staining in late spermatids, making cell morphology useful when checking a reference section (HPA tissue IHC). HPA disregarded nuclear positivity in the CNS as off target; if nuclei dominate, review antibody and detection controls before scoring (HPA tissue IHC; standard IHC practice).
Can this IHC assay distinguish CCT5 isoforms or modification-dependent staining?
CCT5 has 2 annotated isoforms, but the PB9928 caption does not map the antibody epitope, so staining cannot be assigned to either isoform (UniProt P48643; caption PB9928). The annotated chain spans residues 2–541, and reported modifications include phosphoserines at 26, 346 and 539 (UniProt P48643). Without epitope mapping, weak staining alone cannot establish an effect of splicing or phosphorylation (UniProt P48643; standard IHC practice). Compare adjacent sections under identical citrate pH 6 retrieval and detection, then use an independently mapped antibody if isoform discrimination is required (caption PB9928; standard IHC practice).
How can IF help check which cells carry a CCT5 IHC signal?
For a testis IHC finding, multiplex CCT5 with a validated marker of late spermatids and assess signal within the marked cells (HPA tissue IHC; standard IF practice). Inspect tissue autofluorescence before choosing spectrally separated fluorophores, and use single-stain controls to detect channel bleed-through (standard IF practice). CCT5 is cytoplasmic with no transmembrane segment, so an intracellular epitope calls for compatible permeabilisation; the PB9928 epitope itself is unmapped in the supplied caption (UniProt P48643; caption PB9928; standard IF practice). Optimise IF fixation and permeabilisation against preserved morphology, since the reported citrate pH 6, 20-minute retrieval comes from a paraffin-section IHC image (caption PB9928; standard IF practice).
How do I separate weak cytoplasmic CCT5 staining from DAB background?
The PB9928 image used a biotinylated secondary, streptavidin–biotin complex and DAB, so check each detection stage when diffuse brown signal obscures cells (caption PB9928; standard IHC practice). Include a no-primary section and apply a peroxidase block to assess enzyme-driven DAB deposition (standard IHC practice). If background persists, assess endogenous biotin with an appropriate detection control, especially before changing the primary concentration (standard IHC practice; caption PB9928). Compare blocked and unblocked reference sections, titrate the primary around the reported 1 µg/mL, and shorten chromogen development if signal obscures cytoplasm (caption PB9928; standard IHC practice).
How should I score CCT5 IHC across samples with different cell mixtures? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue compartment before scoring, because HPA reports differing CCT5 staining across cell types (HPA tissue IHC; standard IHC practice). For chromogenic sections, record the percentage of positive cells and intensity grades, or calculate an H-score on a 0–300 scale from percentages at each intensity grade (standard IHC practice). Normalise counts to the number of eligible cells or stained area to the measured viable tissue area, rather than whole-section area when necrosis or empty space varies (standard IHC practice). Use the same retrieval, imaging and threshold settings across slides, with a shared reference section to monitor run variation (standard IHC practice).
Which findings support genuine CCT5 positivity rather than tissue or detection artefact?
A credible result shows cellular cytoplasmic signal in preserved tissue, consistent with CCT5 localisation and HPA’s general tissue profile (UniProt P48643; HPA tissue IHC). Check the identity of stained cells: HPA reports high staining in late spermatids and germinal center cells, while some squamous epithelial populations are undetected (HPA tissue IHC). Treat dominant nuclear CNS staining cautiously because HPA disregarded it as off target (HPA tissue IHC). Exclude necrotic regions and section edges with disproportionate colour, then inspect no-primary and peroxidase-control sections for detection artefacts before assigning a positive score (standard IHC practice).
Boster reagents
Best CCT5 / T-complex protein 1 subunit epsilon IHC Antibodies
PB9928 has IHC images from human intestinal cancer, mouse ovary and rat ovary paraffin sections, plus an IF image from U20S cells (PB9928 image captions).
Real IHC data
Anti-TCP1 epsilon/CCT5 Antibody ®
Cat # PB9928
PB9928 has documented tissue IHC in human intestinal cancer, mouse ovary and rat ovary paraffin sections (PB9928 IHC captions). PB9928 also has documented IF in U20S cells (PB9928 IF caption).
Which to pick: For tissue IHC, choose PB9928: its own captions document paraffin sections and a primary antibody concentration of 1 μg/ml; the fixative is unreported (PB9928 IHC captions). For IF/ICC, PB9928 is also listed for both applications and has a U20S cell IF image using 2 μg/ml primary antibody (PB9928 catalog applications; PB9928 IF caption). For work across species, PB9928 lists human, mouse and rat reactivity and shows IHC in all three; it is rabbit hosted, while clonality is unreported (PB9928 catalog reactivity and host; PB9928 IHC captions; PB9928 catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P48643 (TCPE_HUMAN, T-complex protein 1 subunit epsilon).
- Human Protein Atlas. CCT5 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CCT5 subcellular location (ICC-IF): Mainly localized to the mid piece. In addition localized to the principal piece and annulus..
- Human Protein Atlas. CCT5 antibody validation summary (3 antibodies).
- Chaperonin containing TCP1 subunit 5 is a tumor associated antigen of non-small cell lung cancer. Oncotarget 2017 — PMC5609992.
- Increased CCT5 expression is a potential unfavourable factor promoting the growth of nasopharyngeal carcinoma. The Journal of international medical research 2024 — PMC11409311.
- CCT5 maintains mitotic fidelity and promotes early colorectal tumorigenesis. iScience 2026 — PMC13014677.
- Integrated bioinformatics and in vitro validation reveal the role of CCT5 in immune infiltration and the Wnt pathway in colon cancer. Discover oncology 2026 — PMC13260463.
- PubMed PMID:7788527 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15372022 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CCT5 (T-complex protein 1 subunit epsilon). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet PB9928). Validated IHC antibody: PB9928 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Glial cells); negative Esophagus. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human intestinal cancer tissues; fixative not specified (datasheet PB9928); 2. Fixation: Image fixative and duration unreported (datasheet PB9928); 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: Citrate pH 6 , 20 min (datasheet PB9928); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9928); 8. Primary antibody: Rabbit anti-CCT5, 0.5-1μg/ml (datasheet PB9928); 9. Primary incubation: Overnight at 4 °C (datasheet PB9928). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9928); Caveat: CNS nuclear positivity is considered off target (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope impact is undetermined (UniProt); Regulation: Stimulus-dependent regulation is not annotated (UniProt). Evidence: UniProt P48643, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the mid piece. In addition localized to the…, Datasheet PB9928, PMC5609992, PMC11409311. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CCT5/immunohistochemistry-ihc-guide.