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- Table of Contents
NOL11 / Nucleolar protein 11 · Western blot design guide
Design a Western Blot for NOL11
Source-linked NOL11 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NOL11 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
NOL11 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 | ~81.1 kDa | |
| Observed band | 90 kDa | |
| Gel | 5–20% (catalog A12538-1) | |
| 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 | Methylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked NOL11 Western Blot Protocol Options
The A12538-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human A431, human HepG2, human MCF-7 (catalog A12538-1) |
| Gel % | 5–20% (catalog A12538-1) |
| Load | 30 ug; reducing conditions (catalog A12538-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A12538-1) |
| Membrane | nitrocellulose membrane (catalog A12538-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A12538-1) |
| Primary antibody | A12538-1 · 0.5 μg/mL (catalog A12538-1) |
| Primary incubation | overnight at 4°C (catalog A12538-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A12538-1) |
| Secondary incubation | 1.5 hour at RT (catalog A12538-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A12538-1) |
| Detection | ECL (catalog A12538-1) |
Section 2
What Is the Expected NOL11 Western Blot Band Size?
NOL11 is predicted at 81.1 kDa and observed near 90 kDa; the cause of the difference is not established.
| Band near 90 kDa | Empirical NOL11 band in whole-cell lysates; confirm identity with appropriate controls |
| Band near 81.1 kDa | Near the predicted mass of NOL11; identity requires confirmation |
| Multiple bands at different sizes | Isoforms 1 and 2 are possible contributors, but distinct migration is unestablished |
| Weak band in cytosolic fraction | NOL11 is localized to the nucleus and nucleolus |
💡Expected NOL11 appearanceNOL11 has a predicted mass of 81.1 kDa, while antibody QC shows a band near 90 kDa in whole-cell lysates; the difference is unexplained, so confirm band identity with appropriate controls.
| Predicted NOL11 mass | 81.1 kDa by sequence; the observed whole-cell band is near 90 kDa, with no established cause for the difference |
| Isoform 1 | May differ in size from isoform 2; its individual migration is not supplied |
| Isoform 2 | May differ in size from isoform 1; its individual migration is not supplied |
| N6-methyllysine at residue 346 | A modification is annotated, but a visible size shift is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nucleolar NOL11 may be poorly recovered | Check nuclear protein recovery and include a positive lysate control |
| Band higher than expected | The observed 90 kDa band exceeds the 81.1 kDa prediction for an undetermined reason | Compare with a positive control and confirm identity by NOL11 depletion |
| Band lower than expected | Isoform identity or band specificity is uncertain | Check antibody epitope coverage and confirm with NOL11 depletion |
| Multiple bands | Isoforms 1 and 2 may contribute, but their migration is unknown | Compare isoform expression and confirm bands by NOL11 depletion |
| Weak or no signal | Nucleolar NOL11 may be underrepresented after extraction | Assess nuclear extraction and test a positive lysate control |
Experiment evidence
Western blot images showing NOL11 band
A12538-1 · NOL11
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NOL11 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for NOL11 in Western blot, you can use adrenal gland tissue, which HPA scores High.
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 alongside the samples.
⚠️Feasibility: NOL11 is nucleolar; adipose tissue is HPA Not detected, but verify its negative signal by Western blot.
HPA tissue expression evidence for NOL11
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 | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | glial cells | High | 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 → |
| Spleen | cells in red pulp | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Medium | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced NOL11 Western Blot Tips
Deeper troubleshooting and optimisation questions for NOL11, answered from its protein features.
How should NOL11 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 NOL11 isoforms produce bands of different sizes?
Isoforms · Yes. UniProt lists isoforms 1 and 2. In isoform 2, residues 154–169 are replaced and residues 170–719 are missing, so it is much shorter than the 719-residue canonical protein. A smaller band is possible, but the feature record does not establish that either isoform is detected on a given blot.
Check the epitope against the UniProt sequences. Isoform 2 lacks canonical residues 170–719 and has a different sequence at 154–169; an antibody targeting those canonical regions may miss it. UniProt coordinates may differ from antibody or paper numbering, so align sequences before comparing positions.
Check the epitope against the UniProt sequences. Isoform 2 lacks canonical residues 170–719 and has a different sequence at 154–169; an antibody targeting those canonical regions may miss it. UniProt coordinates may differ from antibody or paper numbering, so align sequences before comparing positions.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of NOL11?
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 NOL11 Western blot?
Transfer · The canonical protein is predicted at 81.1 kDa, with an observed band near 90 kDa. Choose transfer conditions that retain proteins in this size range, and check both membrane recovery and residual protein in the gel. The supplied features do not specify a uniquely required transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A12538-1 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 can NOL11 bands be quantified consistently?
Quantitation · Quantify the same verified band across samples, using the observed 90 kDa position as a starting reference. Keep isoform-specific bands separate if detected. NOL11 is nucleolar, so use comparable sample preparation and normalization across lanes; its transcription-dependent interaction with FBL does not by itself establish a change in NOL11 abundance.
Why might NOL11 appear at 90 kDa instead of 81.1 kDa?
Interpretation · The supplied blots show an apparent 90 kDa band, while the canonical sequence predicts 81.1 kDa. The listed methylation and isoform features do not establish the cause of that difference. Use the observed position as a reference and confirm band identity independently.
UniProt lists N6-methyllysine at position 346 of the canonical sequence. That residue is absent from isoform 2 because residues 170–719 are missing. The listed modification alone does not predict a resolvable shift or explain the apparent 90 kDa band. Check numbering conventions before comparing sites across sources.
Consider isoform 2 because it lacks canonical residues 170–719, but first check whether the antibody epitope is retained. The feature record alone cannot identify an unexpected band. Compare its position and detection with the expected canonical band near the observed 90 kDa position, then confirm identity independently.
UniProt lists N6-methyllysine at position 346 of the canonical sequence. That residue is absent from isoform 2 because residues 170–719 are missing. The listed modification alone does not predict a resolvable shift or explain the apparent 90 kDa band. Check numbering conventions before comparing sites across sources.
Consider isoform 2 because it lacks canonical residues 170–719, but first check whether the antibody epitope is retained. The feature record alone cannot identify an unexpected band. Compare its position and detection with the expected canonical band near the observed 90 kDa position, then confirm identity independently.
Boster reagents
NOL11 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-NOL11 Antibody Picoband®
Cat # A12538-1
The catalog reports one anti-NOL11 antibody for Western blotting, A12538-1. Its published product blot shows a band near 90 kDa in human HeLa, A431, HepG2, and MCF-7 whole-cell lysates; the expected size is 81 kDa. No independent validation is supplied.
Which to pick: A12538-1 is the only listed option. It is reported reactive with human samples and has a Western blot image from four human cell lines. Consider the observed 90 kDa band alongside the expected 81 kDa size when interpreting results.
Source: BosterBio NOL11 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9H8H0.
- Human Protein Atlas. NOL11 tissue expression.
- PMC12861072 — target-verified WB comparison
- PMC3420923 — target-verified WB comparison