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NUP133 / Nuclear pore complex protein Nup133 · Western blot design guide
Design a Western Blot for NUP133
Source-linked NUP133 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NUP133 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
NUP133 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 | ~129 kDa | |
| Observed band | ~129 kDa | |
| Gel | 5–20% (catalog A05327-2) | |
| Positive control | Adrenal gland (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 + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked NUP133 Western Blot Protocol Options
The A05327-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human K562, human A549, human MCF-7, rat C6, rat NRK, mouse Neuro-2a, mouse NIH/3T3 (catalog A05327-2) |
| Gel % | 5–20% (catalog A05327-2) |
| Load | 30 ug; reducing conditions (catalog A05327-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05327-2) |
| Membrane | nitrocellulose membrane (catalog A05327-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05327-2) |
| Primary antibody | A05327-2 · 0.5 μg/mL (catalog A05327-2) |
| Primary incubation | overnight at 4°C (catalog A05327-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05327-2) |
| Secondary incubation | 1.5 hour at RT (catalog A05327-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05327-2) |
| Detection | ECL (catalog A05327-2) |
Section 2
What Is the Expected NUP133 Western Blot Band Size?
NUP133 is predicted at 129 kDa and observed at approximately 129 kDa; the supplied evidence establishes no difference between these values.
| Single band near 129 kDa | Matches the predicted mass and the empirical NUP133 band |
| Faint 129 kDa band in whole-cell lysate | Nuclear pore localization may limit recovery in the sampled fraction |
| Doublet near 129 kDa | May reflect phosphorylation at annotated sites; band identity needs confirmation |
| Band near 129 kDa in a nuclear fraction | Consistent with NUP133 localization at the nuclear pore |
💡Expected NUP133 appearanceFull-length NUP133 is predicted at 129 kDa and observed at approximately 129 kDa; confirm band identity with an appropriate control such as NUP133 depletion or immunoprecipitation.
| Predicted full-length mass | Places the expected band near 129 kDa, matching the observed band |
| Phosphoserine at residue 7 | Could affect mobility, but no shift is demonstrated |
| Phosphoserine at residue 15 | Could affect mobility, but no shift is demonstrated |
| Phosphothreonine at residue 28 | Could affect mobility, but no shift is demonstrated |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear pore protein may be poorly recovered in the sampled lysate | Check nuclear fraction recovery and include a positive-control lysate |
| Band higher than expected | Band identity or altered mobility is uncertain | Compare with the 129 kDa reference and test NUP133 depletion |
| Band lower than expected | Possible sample degradation or nonspecific signal | Use protease inhibitors and test NUP133 depletion |
| Multiple bands | Annotated phosphorylation could contribute, but the identities are unproven | Compare phosphatase-treated samples and test NUP133 depletion |
| Weak or no signal | Limited recovery of nuclear pore-associated NUP133 | Check nuclear protein recovery and antibody performance with a positive control |
| Fragments below expected size | Possible sample proteolysis | Prepare samples with protease inhibitors and compare fresh lysate |
Experiment evidence
Western blot images showing NUP133 band
A05327-2 · NUP133
A05327-2 · NUP133
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NUP133 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for NUP133 in Western blot, you can use adrenal gland tissue.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: Adipose tissue is listed as not detected by HPA, but confirm its suitability as a negative control in your lysate.
HPA tissue expression evidence for NUP133
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 | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal 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 → |
| Endometrium | cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Lung | alveolar cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NUP133 Western Blot Tips
Deeper troubleshooting and optimisation questions for NUP133, answered from its protein features.
How should NUP133 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 NUP133 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no isoform-based explanation for multiple bands; check band specificity before assigning additional bands to NUP133.
Which NUP133 modifications matter when interpreting band changes?
PTM · UniProt lists phosphorylation, methylation and acetylation sites, including phosphoserines 480, 489, 493 and 501. These are UniProt sequence coordinates and may differ from antibody or paper numbering. Site presence alone does not show that a modification changes mobility.
Does this guide establish induction of NUP133?
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 NUP133 Western blot?
Transfer · For a protein near 129 kDa, start with wet transfer and verify recovery around 129 kDa using a total-protein stain or marker. Adjust transfer conditions if protein remains in the gel; the supplied features do not specify a validated NUP133 transfer protocol.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05327-2 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 NUP133 bands be quantified?
Quantitation · Measure the band near 129 kDa within a linear exposure range and normalize across matched lanes. Compare equivalent sample fractions: NUP133 is associated with nuclear pores and also localizes to kinetochores during mitosis, so a fraction-specific signal may change without establishing a change in total abundance.
Should NUP133 migrate above its predicted mass?
Interpretation · NUP133 has a predicted mass of 129 kDa, and the supplied Western blots show a band near 129 kDa. Its listed modifications do not, by themselves, establish a visible shift or explain any future difference in apparent mass.
Compare it with the expected band near 129 kDa and check antibody specificity. The record lists one isoform, no signal peptide or propeptide, and no glycosylation sites. Its listed modifications alone do not identify an unexpected band or establish a visible shift.
NUP133 is listed at nuclear pores and, during mitosis, at kinetochores. Keep cell-cycle state and fractionation consistent when comparing blots. A change in a nuclear pore or chromosome-enriched fraction may reflect localization rather than total NUP133 abundance.
Compare it with the expected band near 129 kDa and check antibody specificity. The record lists one isoform, no signal peptide or propeptide, and no glycosylation sites. Its listed modifications alone do not identify an unexpected band or establish a visible shift.
NUP133 is listed at nuclear pores and, during mitosis, at kinetochores. Keep cell-cycle state and fractionation consistent when comparing blots. A change in a nuclear pore or chromosome-enriched fraction may reflect localization rather than total NUP133 abundance.
Boster reagents
NUP133 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-NUP133 Antibody Picoband®
Cat # A05327-2
The catalog reports one anti-NUP133 antibody, A05327-2, for Western blotting. Its product image reports a band near the expected 129 kDa in human, rat, and mouse cell lysates. No publication or independent validation evidence is supplied.
Which to pick: A05327-2 is the only listed option. Its stated reactivity is human and rat; the WB image includes human K562, A549, and MCF-7, rat C6 and NRK, and mouse Neuro-2a and NIH/3T3 lysates. Mouse appears in the image but is not listed under reactivity.
Source: BosterBio NUP133 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8WUM0.
- Human Protein Atlas. NUP133 tissue expression.
- PMC9025798 — target-verified WB comparison
- PMC7447710 — target-verified WB comparison
- PMC8234485 — target-verified WB comparison