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- Table of Contents
CEP164 / Centrosomal protein of 164 kDa · Western blot design guide
Design a Western Blot for CEP164
Source-linked CEP164 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CEP164 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
CEP164 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 | ~164.3 kDa | |
| Observed band | ~164 kDa | |
| Gel | 5–20% (catalog A05971-1) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Adrenal gland (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked CEP164 Western Blot Protocol Options
The A05971-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human K562, human SIHA (catalog A05971-1) |
| Gel % | 5–20% (catalog A05971-1) |
| Load | 30 ug; reducing conditions (catalog A05971-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05971-1) |
| Membrane | nitrocellulose membrane (catalog A05971-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05971-1) |
| Primary antibody | A05971-1 · 0.5 μg/mL (catalog A05971-1) |
| Primary incubation | overnight at 4°C (catalog A05971-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05971-1) |
| Secondary incubation | 1.5 hour at RT (catalog A05971-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05971-1) |
| Detection | ECL (catalog A05971-1) |
Section 2
What Is the Expected CEP164 Western Blot Band Size?
CEP164 is predicted at 164.3 kDa and observed at ~164 kDa; isoforms and phosphorylation are annotated, but distinct migration effects are unproven.
| Band at ~164 kDa | Matches the observed CEP164 band and its 164.3 kDa predicted mass |
| Additional band near ~164 kDa | Could reflect isoform 1 or 2, but their migration is not established |
| Band at a different position | Phosphorylation could affect mobility, but a visible shift is not established |
| Weak band in a whole-cell lysate | CEP164 localizes to centrioles and the nucleus; assess recovery and band identity |
💡Expected CEP164 appearanceCEP164 has a predicted mass of 164.3 kDa and an observed band at ~164 kDa; confirm identity with antibody and sample controls before assigning additional bands to isoforms or phosphorylation.
| UniProt predicted mass | Places the full-length protein at 164.3 kDa, consistent with the observed ~164 kDa band |
| Isoforms 1 and 2 | May differ in size, but their relative masses and migration are not supplied |
| Phosphoserine 186 by ATR and ATM | Could affect mobility; a visible shift is not established |
| Phosphoserine 201 | Could affect mobility; a visible shift is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Poor recovery of centriole-associated or nuclear CEP164 | Check extraction and fraction recovery with compartment markers |
| Band higher than expected | Phosphorylation could affect migration, but the cause is unverified | Compare phosphatase-treated and untreated samples and confirm band identity |
| Band lower than expected | An isoform or degradation is possible; isoform sizes are unknown | Use an independent epitope antibody and fresh lysate to check identity |
| Multiple bands | Isoforms 1 and 2 or phosphorylation are possible, but separation is unproven | Compare independent antibodies and phosphatase-treated samples |
| Weak or no signal | Limited recovery of localized CEP164 | Check loading and enrich the relevant cellular fraction |
Experiment evidence
Western blot images showing CEP164 band
A05971-1 · CEP164
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CEP164 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CEP164 in Western blot, you can use appendix tissue, which HPA lists as a positive sample.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Adrenal gland (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: CEP164 is centrosomal and HPA rates the listed positive tissues as medium, so whole-lysate signal may be modest.
HPA tissue expression evidence for CEP164
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 |
|---|---|---|---|---|
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Colon | glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CEP164 Western Blot Tips
Deeper troubleshooting and optimisation questions for CEP164, answered from its protein features.
How should CEP164 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 CEP164 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. In isoform 2, E at canonical position 469 becomes ERYH, and canonical residues 1242–1250 (REWWRQQRI) become L. These sequence changes could affect band interpretation, but the features do not establish that the isoforms resolve as separate bands.
Which CEP164 phosphorylation sites matter when interpreting a band?
PTM · UniProt lists phosphoserines at canonical positions 186, 201, 566, 1386, 1388, and 1443. ATR and ATM are specified for Ser186. Compare matched samples if testing phosphorylation effects; these sites alone do not predict a visible shift. Antibody or paper numbering may differ from these UniProt coordinates.
Does this guide establish induction of CEP164?
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 CEP164?
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 A05971-1 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 CEP164 signal be quantified across samples?
Quantitation · Compare the ~164 kDa band under matched sample preparation and loading conditions. CEP164 is annotated at centrioles and in the nucleus, so keep the sampled fraction consistent when comparing its signal. The supplied features do not establish that its abundance is constant across conditions.
Why does CEP164 run near 164 kDa?
Interpretation · The supplied apparent band is ~164 kDa, close to CEP164’s predicted 164.3 kDa. Its listed phosphorylation sites and isoforms do not, by themselves, establish a visible shift or explain any mass difference.
CEP164 has DNA damage and DNA repair annotations, interacts with ATR and ATM, and is phosphorylated at UniProt Ser186 by both enzymes. Matched treated and untreated samples can test for a change, but these features do not establish increased total CEP164 or a visible band shift.
Check the ~164 kDa band first. Isoform 2 has sequence changes at canonical positions 469 and 1242–1250, and CEP164 has six listed phosphoserines; neither feature alone identifies an unexpected band. Compare matched samples before assigning a band to an isoform or modification.
Yes. UniProt places CEP164 at the centrosome and centriole, specifically distal appendages of the mature centriole, and also lists the nucleus. Compare equivalent fractions across samples when evaluating band intensity; a change in sampled fraction could change how much CEP164 is recovered.
CEP164 has DNA damage and DNA repair annotations, interacts with ATR and ATM, and is phosphorylated at UniProt Ser186 by both enzymes. Matched treated and untreated samples can test for a change, but these features do not establish increased total CEP164 or a visible band shift.
Check the ~164 kDa band first. Isoform 2 has sequence changes at canonical positions 469 and 1242–1250, and CEP164 has six listed phosphoserines; neither feature alone identifies an unexpected band. Compare matched samples before assigning a band to an isoform or modification.
Yes. UniProt places CEP164 at the centrosome and centriole, specifically distal appendages of the mature centriole, and also lists the nucleus. Compare equivalent fractions across samples when evaluating band intensity; a change in sampled fraction could change how much CEP164 is recovered.
Boster reagents
CEP164 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-CEP164 Antibody Picoband®
Cat # A05971-1
A05971-1 is listed for human CEP164. Its Western blot image shows a band near the expected 164 kDa in human K562 and SIHA whole cell lysates. The supplied evidence covers these tested samples and conditions.
Which to pick: Only A05971-1 is listed. It has a Western blot image using human K562 and SIHA lysates; the supplied image does not establish performance in other specimens.
Source: BosterBio CEP164 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UPV0.
- Human Protein Atlas. CEP164 tissue expression.
- PMC5443223 — target-verified WB comparison