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- Table of Contents
NOP58 / Nucleolar protein 58 · Western blot design guide
Design a Western Blot for NOP58
Source-linked NOP58 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NOP58 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
NOP58 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 | ~59.6 kDa | |
| Observed band | ~70 kDa | |
| Gel | 5–20% (catalog A05070-1) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Ubl conjugation | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked NOP58 Western Blot Protocol Options
The A05070-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human K562, human HEL, human MOLT-4, human U251, human HepG2 (catalog A05070-1) |
| Gel % | 5–20% (catalog A05070-1) |
| Load | 30 ug; reducing conditions (catalog A05070-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05070-1) |
| Membrane | nitrocellulose membrane (catalog A05070-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05070-1) |
| Primary antibody | A05070-1 · 0.5 μg/mL (catalog A05070-1) |
| Primary incubation | overnight at 4°C (catalog A05070-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05070-1) |
| Secondary incubation | 1.5 hour at RT (catalog A05070-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05070-1) |
| Detection | ECL (catalog A05070-1) |
Section 2
What Is the Expected NOP58 Western Blot Band Size?
NOP58 is predicted at 59.6 kDa and observed near 70 kDa in reducing whole-cell lysates; the cause of the difference is not established.
| Band near 70 kDa in whole-cell lysate | Matches the empirical NOP58 band; confirm identity with controls |
| Band near 60 kDa | Near the 59.6 kDa predicted mass; identity requires confirmation |
| Close doublet near the main band | NOP58 has phosphorylation sites, but a mobility difference is unproven |
| Band enriched in a nuclear fraction | Consistent with NOP58 localization in the nucleolus and nucleoplasm |
💡Expected NOP58 appearanceNOP58 has a predicted mass of 59.6 kDa, while antibody QC shows a band near 70 kDa in reducing whole-cell lysates; the cause of the difference is unestablished, so confirm identity with appropriate controls.
| Predicted full-length mass | 59.6 kDa is the sequence-based reference; the observed band is near 70 kDa |
| Phosphothreonine at Thr34 | Present, with no demonstrated effect on apparent band size |
| Phosphoserine at Ser109 | Present, with no demonstrated effect on apparent band size |
| Phosphoserines at Ser304, Ser351, Ser483, Ser502 and Ser514 | Present, with no demonstrated effect on apparent band size |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Poor recovery of nuclear NOP58 | Check nuclear extraction and run a positive whole-cell lysate control |
| Band higher than expected | The observed NOP58 band is near 70 kDa despite a 59.6 kDa prediction; the cause is unestablished | Compare with the reported band and verify identity by NOP58 depletion |
| Band lower than expected | Possible degradation or nonspecific binding | Prepare fresh lysate with protease inhibitors and verify identity by NOP58 depletion |
| Multiple bands | Phosphorylation is annotated, but distinct migrating forms are unproven | Check which bands respond to NOP58 depletion |
| Weak or no signal | Insufficient recovery or loading of nuclear protein | Check loading and nuclear extraction against a positive lysate |
Experiment evidence
Western blot images showing NOP58 band
A05070-1 · NOP58
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NOP58 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for NOP58 in Western blot, you can use adrenal gland lysate, which HPA scores High.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: No tissue is reported as Not detected, so use siRNA knockdown or a KO line for a negative control.
HPA tissue expression evidence for NOP58
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 → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | basal cells | High | Protein (IHC) | HPA → |
| Caudate | neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in molecular layer | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Prostate | glandular cells | Low | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
| Soft tissue | chondrocytes | Low | Protein (IHC) | HPA → |
| Testis | Leydig cells | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
Section 3
Advanced NOP58 Western Blot Tips
Deeper troubleshooting and optimisation questions for NOP58, answered from its protein features.
How should NOP58 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.
Should I expect bands from NOP58 isoforms?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It provides no basis for assigning additional bands to annotated isoforms.
Which NOP58 phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphothreonine at position 34 and phosphoserine at positions 109, 304, 351, 483, 502, and 514. These are UniProt coordinates; antibody or paper numbering may differ. Their presence does not establish a visible shift.
Phosphorylation is documented at seven UniProt positions, but the supplied features do not show whether it changes migration detectably. Do not assign the approximately 70 kDa band to phosphorylation from this record alone.
Phosphorylation is documented at seven UniProt positions, but the supplied features do not show whether it changes migration detectably. Do not assign the approximately 70 kDa band to phosphorylation from this record alone.
Does this guide establish induction of NOP58?
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 NOP58?
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 A05070-1 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.
Which sample fraction is relevant for NOP58 quantitation?
Quantitation · NOP58 localizes mainly to the nucleolus, with a minor nucleoplasmic fraction. Keep the fraction and extraction approach consistent across samples when comparing signal, and state whether measurements represent whole cells or a nuclear fraction.
Why does NOP58 appear near 70 kDa instead of 59.6 kDa?
Interpretation · The reported apparent band is about 70 kDa, while the predicted mass is 59.6 kDa. UniProt lists phosphorylation and a Ubl conjugation keyword, but these features alone do not explain the difference. Confirm the band’s identity before assigning a cause.
The record lists one isoform, no signal peptide or propeptide, and no glycosylation sites. It also lists phosphorylation and Ubl conjugation, without establishing their effects on band migration. Treat additional bands as unidentified until independently verified.
The record lists one isoform, no signal peptide or propeptide, and no glycosylation sites. It also lists phosphorylation and Ubl conjugation, without establishing their effects on band migration. Treat additional bands as unidentified until independently verified.
Boster reagents
NOP58 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-NOP58 Antibody Picoband®
Cat # A05070-1
A05070-1 is a human-reactive anti-NOP58 antibody with a Western blot image from six human whole-cell lysates. The caption reports a band near 70 kDa, versus an expected 60 kDa; evidence for other species is not supplied.
Which to pick: A05070-1 is the only listed option. Its WB image includes HeLa, K562, HEL, MOLT-4, U251, and HepG2 lysates; consider the reported 70 kDa band when interpreting results.
Source: BosterBio NOP58 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9Y2X3.
- Human Protein Atlas. NOP58 tissue expression.
- PMC10612268 — target-verified WB comparison
- PMC13019138 — target-verified WB comparison
- PMC12076867 — target-verified WB comparison