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- Table of Contents
CEP290 / Centrosomal protein of 290 kDa · Western blot design guide
Design a Western Blot for CEP290
Real validated CEP290 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CEP290 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
CEP290 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~290.4 kDa | |
| Gel | 4–12% gradient (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Soft tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated CEP290 Western Blot Protocols
The A01894 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HCT116 (40ug), SGC7901 (40ug), COS-7 (40ug) (catalog A01894) |
| Gel % | 4–12% gradient (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A01894; use the WB datasheet starting dilution (standard starting point) |
| 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 CEP290 Western Blot Band Size?
CEP290 is predicted at 290.4 kDa; its listed features do not demonstrate different migration, and no empirical band size is supplied.
| Band near 290.4 kDa | Consistent with the predicted size of full-length CEP290; confirm its identity |
| Faint band near 290.4 kDa in whole-cell lysate | CEP290 may be poorly recovered from its centrosomal and cytoskeletal locations |
| No band in a soluble fraction | Centrosomal or cytoskeletal CEP290 may remain in the insoluble fraction |
| Stronger band near 290.4 kDa in a cytoskeletal fraction | Consistent with enrichment of localized CEP290; confirm its identity |
💡Expected CEP290 appearanceUniProt predicts full-length CEP290 at 290.4 kDa; no empirical band size is supplied, so confirm a band near that size with ordinary band-identity controls.
| UniProt predicted mass | Places full-length CEP290 at 290.4 kDa by sequence calculation |
| 290386 Da calculated mass | Corresponds to approximately 290.4 kDa for full-length CEP290 |
| Full-length sequence prediction | Provides a size reference without establishing electrophoretic migration |
| Single listed isoform | Provides no evidence for a second isoform band at a different size |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Centrosomal and cytoskeletal CEP290 may be poorly recovered in the tested fraction | Check fraction recovery and include a positive control |
| Band higher than expected | The supplied features do not establish the cause | Confirm identity with CEP290 depletion or a second antibody |
| Band lower than expected | The supplied features do not establish a cleavage product | Check sample integrity and confirm identity with a second antibody |
| Multiple bands | No distinct isoform masses or processing products are supplied to identify them | Check which bands disappear after CEP290 depletion |
| Weak or no signal | Recovery of localized CEP290 may be low | Assess extraction and loading with a positive control |
Experiment evidence
Western blot images showing CEP290 band
A01894 · CEP290
A01894-1 · CEP290
A01894-1 · CEP290
A30603 · CEP290
A30603 · CEP290
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CEP290 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CEP290 in Western blot, you can use adrenal gland tissue.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Soft tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside.
⚠️Feasibility: Soft tissue is HPA not detected, but its Western blot signal should be checked.
HPA tissue expression evidence for CEP290
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | ciliated cells (cell body) | High | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Soft tissue | chondrocytes | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Low | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Low | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced CEP290 Western Blot Tips
Deeper troubleshooting and optimisation questions for CEP290, answered from its protein features.
Where should the full-length CEP290 band appear?
Band shift · CEP290 is 2,479 amino acids long with a predicted mass of 290.4 kDa. Use that value to identify a candidate full-length band, but assess identity with appropriate controls; no empirical apparent band position is supplied.
Does UniProt support multiple CEP290 isoform bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no basis for assigning separate bands to annotated isoforms.
Could modification explain a shifted CEP290 band?
PTM · Ubl conjugation appears among the keywords, but the supplied features list no modified residues or modification coordinates. These features cannot establish whether a modification causes a visible shift or explain a difference from 290.4 kDa.
Does this guide establish induction of CEP290?
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.
What transfer method to use for CEP290 Western blot?
Transfer · CEP290 has a predicted mass of 290.4 kDa. Choose a transfer setup suitable for a protein of that size and check transfer efficiency near 290 kDa. The supplied features do not specify a membrane or transfer settings.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01894 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 CEP290 band intensity be quantified?
Quantitation · Quantify a consistently identified band near the expected full-length size and normalize it to a suitable loading measure. Because the supplied record gives no empirical band position, verify band identity before comparing intensities.
How should unexpected lower CEP290 bands be interpreted?
Interpretation · Do not assign lower bands to annotated isoforms: the supplied record lists only one isoform. Compare bands with an appropriate specificity control before attributing them to CEP290; the supplied features do not identify their cause.
Boster reagents
CEP290 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-CEP290 (F812) Antibody
Cat # A01894
Real WB data
Anti-Centrosomal protein of 290 kDa CEP290 Antibody
Cat # A01894-1
Real WB data
Anti-Centrosomal protein of 290 kDa CEP290 Antibody
Cat # A30603
Three the supplier anti-CEP290 polyclonal antibodies are listed with WB images. A01894 has a caption naming seven cell lysates, 40 µg per lane, and a 1:500 dilution; the other two captions describe only unspecified cells.
Which to pick: Choose A01894 if you need reported rat reactivity or a WB example with named lysates and conditions. A01894-1 and A30603 list human and mouse reactivity, but their WB captions give no specific samples or conditions.
Source: BosterBio CEP290 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O15078.
- Human Protein Atlas. CEP290 tissue expression.
- PMC6245950 — target-verified WB comparison
- PMC10187159 — target-verified WB comparison
- PMC10297203 — target-verified WB comparison