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- Table of Contents
RUVBL2 / RuvB-like 2 · Western blot design guide
Design a Western Blot for RUVBL2
Source-linked RUVBL2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RUVBL2 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
RUVBL2 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 | ~51.2 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Bronchus (IHC candidate; verify WB) +4 more | |
| Negative control | Adrenal gland (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked RUVBL2 Western Blot Protocol Options
The M02246-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | (1) HeLa cell lysate; (2) NIH/3T3 cell lysate (catalog M02246-1) |
| 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 | M02246-1; 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 RUVBL2 Western Blot Band Size?
RUVBL2 is predicted at 51.2 kDa; its isoforms and homohexameric assembly may affect patterns, but no migration effect or empirical band is supplied.
| Band near 51.2 kDa | Consistent with the predicted RUVBL2 monomer; confirm identity with antibody controls |
| Several bands near the monomer region | Isoforms 1 and 2 are annotated, but distinct migration is unproven |
| High-mass band under native conditions | Could reflect annotated homohexameric rings |
| Shifted band of uncertain size | Ser437 phosphorylation is annotated, but a mobility shift is unproven |
💡Expected RUVBL2 appearanceRUVBL2 has a predicted monomer mass of 51.2 kDa, with no empirical band supplied; confirm any band near that size using antibody specificity and appropriate controls.
| Predicted monomer mass | Places the reference band near 51.2 kDa |
| Isoform 1 | Its size relative to isoform 2 is not supplied |
| Isoform 2 | Its size relative to isoform 1 is not supplied |
| Homohexameric rings | May appear at higher mass under native conditions; SDS migration is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | RUVBL2 is mainly nuclear, so extraction may be insufficient | Check nuclear extraction and a positive-control lysate |
| Band higher than expected | Native RUVBL2 can form homohexameric rings; the identity of this band is uncertain | Compare native and denaturing samples and verify antibody specificity |
| Band lower than expected | An alternative isoform is possible, but its mass is unknown | Use isoform-aware controls and verify band identity |
| Multiple bands | Isoforms 1 and 2 are annotated; distinct migration is unproven | Check antibody specificity and compare isoform expression |
| Weak or no signal | The sampled fraction may contain little mainly nuclear RUVBL2 | Compare nuclear and whole-cell fractions with loading controls |
Experiment evidence
Western blot images showing RUVBL2 band
M02246-1 · RUVBL2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RUVBL2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RUVBL2 in Western blot, you can use bronchus tissue, which HPA scores High.
Positive control: Bronchus (IHC candidate; verify WB)
Negative control: Adrenal gland (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: An intracellular signal should be feasible in tissue lysate; HPA reports no detection in adrenal gland for a negative control.
HPA tissue expression evidence for RUVBL2
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 |
|---|---|---|---|---|
| Bronchus | ciliated cells (cell body) | High | Protein (IHC) | HPA → |
| Placenta | cytotrophoblasts | High | Protein (IHC) | HPA → |
| Testis | pachytene spermatocytes | High | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Cervix | glandular cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Not detected | Protein (IHC) | HPA → |
| Colon | endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RUVBL2 Western Blot Tips
Deeper troubleshooting and optimisation questions for RUVBL2, answered from its protein features.
What band size should I expect for RUVBL2?
Band shift · The supplied predicted mass is 51.2 kDa for the 463-residue canonical sequence. No observed band size is available, so use 51.2 kDa as a reference, not a guaranteed migration position.
Could RUVBL2 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 1–45 of the canonical sequence, so it may migrate differently. Whether both appear depends on their abundance and whether the antibody recognizes the sequence retained in isoform 2.
Which modifications matter when interpreting RUVBL2 bands?
PTM · The supplied features list N-acetylalanine at canonical position 2 and phosphoserine at canonical position 437. Neither feature alone establishes a visible shift or explains a difference from 51.2 kDa. Check which isoform the antibody detects before comparing residue numbers across sources.
Phosphoserine is listed at canonical residue 437, but the feature alone does not show that phosphorylation changes migration detectably. If investigating a shift, compare conditions with an appropriate phosphatase-treated sample and keep the canonical numbering explicit.
Phosphoserine is listed at canonical residue 437, but the feature alone does not show that phosphorylation changes migration detectably. If investigating a shift, compare conditions with an appropriate phosphatase-treated sample and keep the canonical numbering explicit.
Does this guide establish induction of RUVBL2?
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 RUVBL2?
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 M02246-1 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 should I quantify RUVBL2 across samples?
Quantitation · Decide whether the measurement is total RUVBL2 or an individual band. UniProt lists two isoforms and places RUVBL2 mainly in the nucleus, with additional cytoplasmic and membrane localization. Use comparable sample fractions and an antibody whose isoform coverage is known.
How should I interpret unexpected RUVBL2 bands?
Interpretation · Consider the isoform 2 deletion of canonical residues 1–45 when assessing a lower band. RUVBL2 also forms homohexameric rings and can assemble with RUVBL1, but these features alone do not identify an extra band. Compare antibody recognition and sample preparation before assigning its identity.
Boster reagents
RUVBL2 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-Reptin / RUVBL2 Rabbit Monoclonal Antibody
Cat # M02246-1
The catalog reports one anti-RUVBL2 rabbit monoclonal antibody for Western blot, M02246-1, with reported human and mouse reactivity. Its WB image uses HeLa and NIH/3T3 cell lysates; broader validation evidence is not supplied.
Which to pick: M02246-1 is the only listed option. Its WB image includes HeLa and NIH/3T3 lysates. Check the reported reactivity and these tested samples against your planned experiment.
Source: BosterBio RUVBL2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9Y230.
- Human Protein Atlas. RUVBL2 tissue expression.
- PMC11066559 — target-verified WB comparison
- PMC11629685 — target-verified WB comparison
- PMC6764127 — target-verified WB comparison