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- Table of Contents
AURKB / Aurora kinase B · Western blot design guide
Design a Western Blot for AURKB
Real validated AURKB Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-AURKB 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
AURKB 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 | ~39.3 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (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 Hallmark membership | |
| Isoform | 5 isoform(s) |
Section 1
Real Curated AURKB Western Blot Protocols
The A00762 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | H1792 (40ug), HCT116 (40ug), LOVO (40ug) (catalog A00762) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A00762; 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 AURKB Western Blot Band Size?
AURKB is predicted at 39.3 kDa; isoforms could affect migration, but no empirical band or distinct isoform pattern is supplied.
| Band near 39.3 kDa | Consistent with the predicted AURKB mass; confirm identity with controls |
| Several bands at different positions | Could reflect AURKB splice isoforms, though distinct migration is unproven |
| Band below 39.3 kDa | Could be an AURKB isoform; its mass is not supplied |
| Band above 39.3 kDa | Could be an AURKB isoform; its mass is not supplied |
💡Expected AURKB appearanceUniProt predicts AURKB at 39.3 kDa, but no empirical band size or isoform migration pattern is supplied; verify any candidate band with identity controls.
| UniProt predicted mass | Places the reference band near 39.3 kDa |
| Splice isoform 1 | Its individual mass and migration relative to other isoforms are unknown |
| Splice isoforms 2 and 3 | Their individual masses and migration relative to other isoforms are unknown |
| Splice isoforms 4 and 5 | Their individual masses and migration relative to other isoforms are unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear and chromosome-associated AURKB may be poorly recovered | Check extraction of the nuclear fraction and use a positive-control lysate |
| Band higher than expected | Isoform identity or nonspecific binding is unresolved | Compare with an AURKB depletion control |
| Band lower than expected | An isoform or degradation could account for the band | Check sample integrity and compare with an AURKB depletion control |
| Multiple bands | Five annotated isoforms exist, but their migration is unknown | Identify AURKB-dependent bands with a depletion control |
| Weak or no signal | AURKB may be poorly recovered from nuclear material | Verify nuclear extraction and test a positive-control lysate |
Experiment evidence
Western blot images showing AURKB band
A00762 · AURKB
A00762T232 · AURKB
M00762-1 · AURKB
A00762-1 · AURKB
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for AURKB Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for AURKB in Western blot, you can use appendix tissue lysate, which HPA scores High.
Positive control: Appendix (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: AURKB is intracellular, and HPA scores adipose tissue Not detected, though a negative lysate may still need validation.
HPA tissue expression evidence for AURKB
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 | endocrine cells | High | Protein (IHC) | HPA → |
| Colon | endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | endocrine cells | High | Protein (IHC) | HPA → |
| Rectum | endocrine cells | High | Protein (IHC) | HPA → |
| Skin | cells in basal layer | 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 → |
| Caudate | glial cells | 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 AURKB Western Blot Tips
Deeper troubleshooting and optimisation questions for AURKB, answered from its protein features.
Where should the main AURKB band appear?
Band shift · The canonical sequence predicts 39.3 kDa. No observed band position is supplied, so use 39.3 kDa as a starting reference, not a confirmed migration position. The listed modifications alone do not establish a visible shift.
Could AURKB isoforms produce additional bands?
Isoforms · Five isoforms are listed. Isoform 4 lacks canonical residues 1–41, isoform 2 replaces residues 86–134, and isoform 3 lacks residues 142–344. These sequence differences could affect band size or antibody recognition; the supplied features do not establish which isoforms are present in a sample.
Which phosphorylation sites should a phospho-AURKB assay consider?
PTM · UniProt lists phosphothreonine at 35, 64 and 232, and phosphoserine at 62 and 227. Thr232 is annotated as autophosphorylated. Match the antibody’s stated site to these UniProt coordinates, checking its numbering convention before interpreting a phospho-specific band.
Yes. UniProt lists N6-acetyllysine at position 215. If measuring this modification, use an assay that distinguishes acetylated AURKB from total AURKB; the feature alone does not establish its occupancy or a visible mass shift.
Measure the phospho-specific signal and total AURKB signal separately, then compare phospho signal with total AURKB for the same samples. UniProt lists five phosphorylation sites, but their presence does not show how much protein is phosphorylated in a given sample.
Yes. UniProt lists N6-acetyllysine at position 215. If measuring this modification, use an assay that distinguishes acetylated AURKB from total AURKB; the feature alone does not establish its occupancy or a visible mass shift.
Measure the phospho-specific signal and total AURKB signal separately, then compare phospho signal with total AURKB for the same samples. UniProt lists five phosphorylation sites, but their presence does not show how much protein is phosphorylated in a given sample.
Does this guide establish induction of AURKB?
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 AURKB?
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 A00762 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 AURKB be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
How should unexpected AURKB bands be interpreted?
Interpretation · First compare them with the 39.3 kDa canonical prediction and the listed isoform sequence changes. A band difference could warrant checking which isoforms an antibody can recognize. Phosphorylation and Lys215 acetylation are annotated, but those features alone cannot identify an unexpected band or explain an apparent mass difference.
Boster reagents
AURKB 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-AURKB/C (K197) Antibody
Cat # A00762
Real WB data
Anti-Phospho-ARK-2 (T232) AURKB Antibody
Cat # A00762T232
Real WB data
Anti-Aurora B AURKB Rabbit Monoclonal Antibody
Cat # M00762-1
Real WB data
Anti-AURKB/C (K197) Antibody
Cat # A00762-1
All four listed antibodies have WB images. A00762 and A00762-1 show AURKB/C (K197) blots of named whole-cell lysates. M00762-1 shows an approximately 39 kDa band in human lysates. A00762T232 shows phospho-Thr232 detection in treated COS7 lysate with phosphopeptide blocking. Evidence is limited to the shown conditions.
Which to pick: For phospho-Thr232, choose A00762T232. For Aurora B in human lysates, M00762-1 has a documented 39 kDa band. A00762 and A00762-1 list human, mouse, and rat reactivity; compare their pictured lysates with your planned sample.
Source: BosterBio AURKB gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96GD4.
- Human Protein Atlas. AURKB tissue expression.
- PMC12379206 — target-verified WB comparison
- PMC8866333 — target-verified WB comparison
- PMC11132018 — target-verified WB comparison