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- Table of Contents
CCT5 / T-complex protein 1 subunit epsilon · Western blot design guide
Design a Western Blot for CCT5
Source-linked CCT5 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CCT5 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
CCT5 Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~59.7 kDa | |
| Observed band | ~60 kDa | |
| Gel | 5–20% (catalog PB9928) | |
| Positive control | Caudate (IHC candidate; verify WB) +4 more | |
| Negative control | Esophagus (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked CCT5 Western Blot Protocol Options
The PB9928 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human MCF-7, human SiHa, human A549, human 293T, human CACO-2, human PC-3, human U20S, rat testis, mouse testis, mouse liver (catalog PB9928) |
| Gel % | 5–20% (catalog PB9928) |
| Load | 30 ug; reducing conditions (catalog PB9928) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog PB9928) |
| Membrane | nitrocellulose membrane (catalog PB9928) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog PB9928) |
| Primary antibody | PB9928 · 0.5 μg/mL (catalog PB9928) |
| Primary incubation | overnight at 4°C (catalog PB9928) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog PB9928) |
| Secondary incubation | 1.5 hour at RT (catalog PB9928) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog PB9928) |
| Detection | ECL (catalog PB9928) |
Section 2
What Is the Expected CCT5 Western Blot Band Size?
CCT5 is predicted at 59.7 kDa and observed at ~60 kDa; annotated isoforms have no established distinct band sizes.
| Single band at ~60 kDa | Consistent with the observed CCT5 band and its 59.7 kDa predicted mass |
| Single band despite two annotated isoforms | Isoforms 1 and 2 need not form resolvable bands |
| Additional band at a different mass | Could reflect an isoform, but its identity requires validation |
| Higher band under insufficiently denaturing conditions | Could reflect retained TRiC assembly rather than a CCT5 monomer |
💡Expected CCT5 appearanceCCT5 has a predicted mass of 59.7 kDa and an observed band at ~60 kDa; confirm band identity with appropriate controls, especially if additional bands appear.
| UniProt predicted mass of 59.7 kDa | Provides the reference size for CCT5 |
| UniProt sequence mass of 59,671 Da | Corresponds to the approximately 60 kDa monomer reference |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 2 | Its size relative to isoform 1 is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | CCT5 signal may be below detection in the tested cytoplasmic sample | Check lysate loading and use a CCT5-positive lysate control |
| Band higher than expected | TRiC assembly may be incompletely dissociated | Check denaturation conditions and confirm band identity |
| Band lower than expected | The listed isoforms have no supplied migration assignments | Confirm identity with an independent antibody or CCT5 depletion |
| Multiple bands | Isoforms 1 and 2 are annotated, but distinct bands are not established | Validate each band with CCT5 depletion or an independent antibody |
| Weak or no signal | CCT5 abundance or recovery may be low in the tested sample | Check loading, transfer and a CCT5-positive control |
Experiment evidence
Western blot images showing CCT5 band
PB9928 · CCT5
PB9928 · CCT5
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CCT5 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CCT5 in Western blot, you can use caudate tissue, which has high HPA expression.
Positive control: Caudate (IHC candidate; verify WB)
Negative control: Esophagus (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: CCT5 is cytoplasmic, so tissue lysates are suitable; HPA reports esophagus as not detected, although a weak signal remains possible.
HPA tissue expression evidence for CCT5
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| 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 → |
Lower expression tissues · IHC evidence, not confirmed WB-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 Western Blot Tips
Deeper troubleshooting and optimisation questions for CCT5, answered from its protein features.
How should CCT5 band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Could CCT5 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 1–38 and 56–110 of the canonical sequence, so it could migrate differently. Check whether the antibody recognizes a region retained in isoform 2 before interpreting an additional band.
Which CCT5 phosphorylation sites matter for band interpretation?
PTM · UniProt lists phosphoserine at canonical positions 26, 346, and 539. Position 26 falls within a region missing from isoform 2; positions 346 and 539 are retained, though their numbering changes in that isoform. These sites do not establish that a visible band shift will occur.
UniProt lists N-acetylalanine at canonical position 2, within the region missing from isoform 2. The feature alone does not show that acetylation produces a separate detectable band.
UniProt lists N-acetylalanine at canonical position 2, within the region missing from isoform 2. The feature alone does not show that acetylation produces a separate detectable band.
Does this guide establish induction of CCT5?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
How should transfer be checked for CCT5?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the PB9928 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should CCT5 be quantified across cell fractions?
Quantitation · CCT5 is listed in the cytoplasm and at the centrosome. Compare like fractions across samples and keep fraction preparation consistent; a change in a fraction's CCT5 signal need not represent the same change in total cellular CCT5.
Why is the CCT5 band near 60 kDa?
Interpretation · The observed band near 60 kDa is close to the 59.7 kDa predicted mass. The listed modifications alone do not establish a visible shift or explain any small difference between apparent and calculated mass.
CCT5 is a component of the hexadecameric TRiC complex. A high-mass band alone cannot establish that intact TRiC was detected. Interpret it alongside the expected CCT5 band near 60 kDa and the sample preparation conditions.
Check whether a lower band is consistent with isoform 2, which lacks canonical residues 1–38 and 56–110. The antibody must recognize a retained region for that interpretation. The supplied features do not identify any cleavage product.
CCT5 is a component of the hexadecameric TRiC complex. A high-mass band alone cannot establish that intact TRiC was detected. Interpret it alongside the expected CCT5 band near 60 kDa and the sample preparation conditions.
Check whether a lower band is consistent with isoform 2, which lacks canonical residues 1–38 and 56–110. The antibody must recognize a retained region for that interpretation. The supplied features do not identify any cleavage product.
Boster reagents
CCT5 Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-TCP1 epsilon/CCT5 Antibody Picoband®
Cat # PB9928
PB9928 is a rabbit polyclonal anti-CCT5 antibody with reported human, mouse, and rat reactivity. Its Western blot caption reports an approximately 60 kDa band in tested human cell lysates and rat and mouse tissue lysates. Evidence is limited to the supplied product caption.
Which to pick: PB9928 is the only listed option and has a Western blot image. Its caption includes human cell lysates, rat testis, and mouse testis and liver, with a reported band near 60 kDa. Match your sample and conditions to those examples.
Source: BosterBio CCT5 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P48643.
- Human Protein Atlas. CCT5 tissue expression.
- PMC11409311 — target-verified WB comparison
- PMC5609992 — target-verified WB comparison
- PMC9043132 — target-verified WB comparison