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- Table of Contents
RBM15 / RNA-binding protein 15 · Western blot design guide
Design a Western Blot for RBM15
Real validated RBM15 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RBM15 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
RBM15 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 | ~107.2 kDa | |
| Observed band | ~107 kDa | |
| Gel | 8% (catalog A04395) | |
| Positive control | Cerebellum (IHC candidate; verify WB) +4 more | |
| Negative control | Soft 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 | 4 isoform(s) |
Section 1
Real Curated RBM15 Western Blot Protocols
The A04395 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human 293T, rat C6, mouse NIH/3T3 (catalog A04395) |
| Gel % | 8% (catalog A04395) |
| Load | 30 ug; reducing conditions (catalog A04395) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04395) |
| Membrane | nitrocellulose membrane (catalog A04395) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A04395) |
| Primary antibody | A04395 · 0.5 μg/mL (catalog A04395) |
| Primary incubation | overnight at 4°C (catalog A04395) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A04395) |
| Secondary incubation | 1.5 hour at RT (catalog A04395) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04395) |
| Detection | ECL (catalog A04395) |
Section 2
What Is the Expected RBM15 Western Blot Band Size?
RBM15 is predicted at 107.2 kDa and observed at ~107 kDa; the small difference has no established cause in the supplied evidence.
| Band at ~107 kDa | Matches the reported RBM15 band and its 107.2 kDa predicted mass. |
| Additional bands at different positions | Could reflect RBM15 isoforms 1, 2, 3, or 4; band identities require confirmation. |
| Close doublet near ~107 kDa | Could reflect differently phosphorylated RBM15; a visible shift is not established. |
| Weak band in a cytoplasmic fraction | Consistent with RBM15 localization mainly in nuclear compartments. |
💡Expected RBM15 appearanceRBM15 is predicted at 107.2 kDa, and antibody QC reports a ~107 kDa band in reducing whole-cell lysates; confirm identity with appropriate band-identity controls.
| UniProt predicted mass | The 107.2 kDa prediction closely matches the observed ~107 kDa band. |
| Isoforms 1 and 2 | Sequence differences could affect size, but their relative masses are unspecified. |
| Isoforms 1 and 3 | Sequence differences could affect size, but their relative masses are unspecified. |
| Isoforms 1 and 4 | Sequence differences could affect size, but their relative masses are unspecified. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear RBM15 may be poorly recovered during extraction. | Check nuclear extraction and compare with a nuclear-enriched fraction. |
| Band higher than expected | A documented RBM15 size-changing mechanism is not established. | Compare with the ~107 kDa reference band and verify identity by RBM15 depletion. |
| Band lower than expected | An isoform or protein breakdown could be responsible. | Use fresh lysate and test whether RBM15 depletion removes the band. |
| Multiple bands | RBM15 has four annotated isoforms, but distinct band positions are unverified. | Check which bands disappear after RBM15 depletion. |
| Weak or no signal | Nuclear RBM15 may be underrepresented in the sample. | Check loading and compare whole-cell with nuclear-enriched lysate. |
| Fragments below expected size | Protein breakdown during sample preparation is possible. | Prepare fresh lysate with protease inhibitors and verify fragment identity by RBM15 depletion. |
Experiment evidence
Western blot images showing RBM15 band
A04395 · RBM15
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RBM15 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RBM15 in Western blot, you can use cerebellum lysate, which has high HPA expression.
Positive control: Cerebellum (IHC candidate; verify WB)
Negative control: Soft tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: RBM15 is nuclear, so nuclear-enriched lysate may improve detection.
HPA tissue expression evidence for RBM15
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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
| Kidney | collecting ducts | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Soft tissue | fibroblasts | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Low | Protein (IHC) | HPA → |
| Breast | glandular cells | Low | Protein (IHC) | HPA → |
| Endometrium | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced RBM15 Western Blot Tips
Deeper troubleshooting and optimisation questions for RBM15, answered from its protein features.
Where should the main RBM15 band appear?
Band shift · RBM15 has a predicted mass of 107.2 kDa, and the supplied observed band is about 107 kDa. The listed modifications alone do not establish a visible shift or explain any difference between calculated and apparent mass.
Could RBM15 isoforms produce different bands?
Isoforms · Four isoforms are listed. Relative to the canonical sequence, isoform 4 lacks residues 1–44; isoform 3 replaces residues 955–977; and isoforms 2 and 4 replace residues 956–977. These changes could affect apparent size or antibody recognition. Check whether the antibody epitope is retained in the isoform of interest.
Which RBM15 phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphoserine at 109, 179, 208, 210, 253, 257, 259, 292, 294, 365, 622, 656, 670, 674, 700, 741, 765, 767, 781 and 935; phosphotyrosine at 266; and phosphothreonine at 568. These are UniProt canonical sequence coordinates, which may differ from antibody or paper numbering. Site presence does not establish a visible band shift.
UniProt lists N6-acetyllysine at canonical residue 450 and alternate asymmetric dimethylarginine or omega-N-methylarginine at canonical residue 578, both methylation forms attributed to PRMT1. If an antibody targets either region, check its stated modification specificity. These annotations do not establish a detectable mobility change.
UniProt lists N6-acetyllysine at canonical residue 450 and alternate asymmetric dimethylarginine or omega-N-methylarginine at canonical residue 578, both methylation forms attributed to PRMT1. If an antibody targets either region, check its stated modification specificity. These annotations do not establish a detectable mobility change.
Does this guide establish induction of RBM15?
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 RBM15?
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 A04395 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 RBM15 bands be quantified across samples?
Quantitation · Quantify the same defined band or bands across samples. RBM15 has four annotated isoforms and multiple modified residues, so document whether the measurement includes one band or several. Use the same fraction and antibody for each comparison; the feature list alone cannot identify every detected band.
Which fraction should be checked for RBM15?
Interpretation · RBM15 is listed in nuclear speckles, nucleoplasm, nuclear envelope and nuclear membrane. Include a nuclear fraction when assessing a weak or absent signal, and compare fractions consistently across samples.
First compare them with the approximately 107 kDa observed band and the four annotated isoforms. Isoform 4 lacks canonical residues 1–44, while isoforms 2, 3 and 4 have C-terminal sequence changes. Check whether the antibody epitope is present in each candidate isoform. The listed modifications alone cannot assign an unexpected band.
First compare them with the approximately 107 kDa observed band and the four annotated isoforms. Isoform 4 lacks canonical residues 1–44, while isoforms 2, 3 and 4 have C-terminal sequence changes. Check whether the antibody epitope is present in each candidate isoform. The listed modifications alone cannot assign an unexpected band.
Boster reagents
RBM15 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-RBM15 Antibody Picoband®
Cat # A04395
A04395 is a rabbit polyclonal anti-RBM15 antibody with a Western blot image showing an approximately 107 kDa band in human 293T, rat C6, and mouse NIH/3T3 whole-cell lysates. No publication evidence is supplied.
Which to pick: A04395 is the only listed option. Its reported reactivity covers human, mouse, and rat, and its WB image documents those three cell lines using 30 µg lysate per lane and 0.5 µg/mL primary antibody.
Source: BosterBio RBM15 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96T37.
- Human Protein Atlas. RBM15 tissue expression.
- PMC12147389 — target-verified WB comparison
- PMC12586852 — target-verified WB comparison
- PMC11214523 — target-verified WB comparison