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- Table of Contents
TTC5 / Tetratricopeptide repeat protein 5 · Western blot design guide
Design a Western Blot for TTC5
Source-linked TTC5 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TTC5 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
TTC5 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 | ~48.9 kDa | |
| Observed band | 68 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Kidney (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Band identity controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked TTC5 Western Blot Protocol Options
The A13808 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | RAW264.7 cell lysate (catalog A13808) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A13808 · (A) 0.5, (B) 1 and (C) 2 μg/mL (catalog A13808) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected TTC5 Western Blot Band Size?
TTC5 is predicted at 48.9 kDa, while antibody QC reports 68 kDa; the cause of the difference is not established.
| Band near 68 kDa | Empirical TTC5 antibody QC band; confirm identity with a knockdown control |
| Band near 48.9 kDa | Position predicted from the full-length TTC5 sequence |
| Closely spaced doublet | Could reflect phosphorylation at Ser-203 or Ser-221; test with phosphatase |
| Weak nuclear band with a stronger cytoplasmic band | Could reflect the reported localization of unphosphorylated TTC5 |
💡Expected TTC5 appearanceTTC5 has a predicted mass of 48.9 kDa, while antibody QC reports a 68 kDa band in cell lysate; the cause of the difference is unestablished, so confirm identity by knockdown.
| Predicted full-length mass | Places unmodified TTC5 near 48.9 kDa by sequence calculation |
| Phosphoserine at Ser-203 | May alter mobility, but no band shift is demonstrated |
| Phosphoserine at Ser-221 | May alter mobility, but no band shift is demonstrated |
| Phosphorylation at Ser-203 and Ser-221 | Could contribute to closely spaced forms; it does not establish the cause of the 68 kDa band |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | The reported 68 kDa band exceeds the 48.9 kDa predicted mass for an unestablished reason | Confirm TTC5 identity by knockdown and compare with the reported 68 kDa band |
| Band lower than expected | Identity or sample integrity is uncertain; no cleavage feature is listed | Compare with a TTC5 knockdown and check sample integrity |
| Multiple bands | Phosphorylation states are possible, but distinct bands are not demonstrated | Compare phosphatase-treated and untreated samples and use a TTC5 knockdown |
| Weak or no signal | TTC5 distribution includes nucleus, cytoplasm, vesicles and mitochondrial matrix | Check whole-cell lysate and relevant fractions with a loading control |
| No band in lysate | The sampled fraction may contain little TTC5 | Test whole-cell lysate and relevant subcellular fractions |
Experiment evidence
Western blot images showing TTC5 band
A13808 · TTC5
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TTC5 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TTC5 in Western blot, you can use kidney tissue, which HPA scores as high for TTC5.
Positive control: Kidney (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside TTC5.
⚠️Feasibility: HPA reports no detectable TTC5 in adipose tissue, but confirm its absence in your lysate.
HPA tissue expression evidence for TTC5
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 |
|---|---|---|---|---|
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TTC5 Western Blot Tips
Deeper troubleshooting and optimisation questions for TTC5, answered from its protein features.
How should TTC5 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 annotated TTC5 isoforms explain multiple bands?
Isoforms · Only one isoform is supplied, with no alternative sequence. The supplied features therefore provide no isoform-based explanation for multiple bands; verify additional bands before assigning them to TTC5.
Which TTC5 phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphoserine at Ser-203 by ATM and Ser-221 by CHEK2. These are UniProt sequence coordinates; antibody or paper numbering may differ. The listed sites alone do not establish a visible band shift.
The supplied localization note links phosphorylation at UniProt Ser-203 to nuclear localization. Compare equivalent fractions when assessing condition-dependent signal: a change in nuclear TTC5 may reflect redistribution rather than a change in total abundance.
The supplied localization note links phosphorylation at UniProt Ser-203 to nuclear localization. Compare equivalent fractions when assessing condition-dependent signal: a change in nuclear TTC5 may reflect redistribution rather than a change in total abundance.
Does this guide establish induction of TTC5?
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 TTC5?
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 A13808 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.
What should be controlled when quantifying TTC5 across conditions?
Quantitation · Keep the sampled compartment consistent. TTC5 is listed in the nucleus and cytoplasm, and phosphorylation at UniProt Ser-203 is linked to nuclear localization. A nuclear-only measurement may change even if total TTC5 does not.
Why might TTC5 appear at 68 kDa instead of 48.9 kDa?
Interpretation · TTC5 has a predicted mass of 48.9 kDa, while the supplied Western blot observation is 68 kDa. Its listed phosphorylation sites do not establish the cause of this difference. Use the observed band as a reference, but verify its identity before attributing the difference to a modification.
The supplied interaction annotation says TTC5 augments nuclear JMY–p300/EP300 association in response to DNA damage, based on similarity. It does not establish increased TTC5 abundance. Compare TTC5 signal across conditions without assuming that DNA damage induces expression.
Start with the supplied 48.9 kDa predicted mass and 68 kDa observed band. One isoform and two phosphorylation sites are listed, but these features do not establish the identity or cause of another band. Verify band identity before assigning it to an isoform or modification.
The supplied interaction annotation says TTC5 augments nuclear JMY–p300/EP300 association in response to DNA damage, based on similarity. It does not establish increased TTC5 abundance. Compare TTC5 signal across conditions without assuming that DNA damage induces expression.
Start with the supplied 48.9 kDa predicted mass and 68 kDa observed band. One isoform and two phosphorylation sites are listed, but these features do not establish the identity or cause of another band. Verify band identity before assigning it to an isoform or modification.
Boster reagents
TTC5 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-TTC5 Antibody
Cat # A13808
The catalog reports A13808, an anti-TTC5 antibody with stated human and mouse reactivity. Its Western blot image shows RAW264.7 cell lysate tested at 0.5, 1, and 2 μg/mL. No publication evidence or human sample image is supplied.
Which to pick: A13808 is the only listed option. Its RAW264.7 Western blot image provides a tested sample and concentration range; human reactivity is listed, but no human Western blot example is supplied.
Source: BosterBio TTC5 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8N0Z6.
- Human Protein Atlas. TTC5 tissue expression.
- PMC13370610 — target-verified WB comparison
- PMC5335694 — target-verified WB comparison
- PMC11570694 — target-verified WB comparison