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- Table of Contents
RBM3 / RNA-binding protein 3 · Western blot design guide
Design a Western Blot for RBM3
Source-linked RBM3 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RBM3 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
RBM3 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 | ~17.2 kDa | |
| Observed band | ~22 kDa | |
| Gel | 12% (catalog A03104-2) | |
| Positive control | Nasopharynx (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 + Methylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked RBM3 Western Blot Protocol Options
The A03104-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human MCF-7, human Raji (catalog A03104-2) |
| Gel % | 12% (catalog A03104-2) |
| Load | 30 ug; reducing conditions (catalog A03104-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A03104-2) |
| Membrane | nitrocellulose membrane (catalog A03104-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A03104-2) |
| Primary antibody | A03104-2 · 0.5 μg/mL (catalog A03104-2) |
| Primary incubation | overnight at 4°C (catalog A03104-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A03104-2) |
| Secondary incubation | 1.5 hour at RT (catalog A03104-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A03104-2) |
| Detection | ECL (catalog A03104-2) |
Section 2
What Is the Expected RBM3 Western Blot Band Size?
RBM3 is predicted at 17.2 kDa and observed near 22 kDa; the cause of the difference is not established.
| Band near 22 kDa | Empirical RBM3 band in whole-cell lysates; confirm identity with controls |
| Band near 17.2 kDa | Near the predicted RBM3 mass; identity still requires confirmation |
| Close doublet near the RBM3 band | May reflect different modification states, but visible separation is unproven |
| No band in a cytoplasmic fraction | RBM3 may be present in the nucleus |
💡Expected RBM3 appearanceRBM3 has a predicted mass of 17.2 kDa and an observed band near 22 kDa in whole-cell lysates; the difference is unexplained, so use ordinary band-identity controls.
| UniProt predicted mass | 17.2 kDa is the sequence-based reference; the observed band is near 22 kDa |
| Arginine methylation at residue 47 | is annotated, but a detectable migration shift is not established |
| Alternative arginine methylation at residue 105 | is annotated, but its effect on band position is not established |
| Phosphoserine at residue 147 | is annotated, but a distinct band is not established |
| Phosphotyrosine at residue 155 | is annotated, but a distinct band is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | RBM3 may be below detection in the sample | Check loading and antibody performance against a positive whole-cell lysate |
| Band higher than expected | The observed RBM3 band is near 22 kDa versus a predicted 17.2 kDa; the cause is unestablished | Compare with the documented band and confirm identity by knockdown or another antibody |
| Band lower than expected | A smaller band has no identified processing explanation in the supplied features | Check sample integrity and confirm band identity |
| Multiple bands | Annotated modification states could contribute, but distinct migration is unproven | Confirm which bands track with RBM3 using knockdown or another antibody |
| Weak or no signal | RBM3 may be missed in a fraction that excludes its nuclear or cytoplasmic pool | Compare nuclear, cytoplasmic, and whole-cell samples with loading controls |
Experiment evidence
Western blot images showing RBM3 band
A03104-2 · RBM3
M03104 · RBM3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RBM3 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RBM3 in Western blot, you can use nasopharynx tissue lysate, which has high HPA expression.
Positive control: Nasopharynx (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: Nasopharynx and adipose tissue offer a feasible positive–negative pair, but confirm the contrast by Western blot.
HPA tissue expression evidence for RBM3
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 |
|---|---|---|---|---|
| Nasopharynx | basal cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Medium | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Medium | 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RBM3 Western Blot Tips
Deeper troubleshooting and optimisation questions for RBM3, answered from its protein features.
How should RBM3 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 RBM3 isoforms explain multiple bands?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands. Assess additional bands separately rather than assigning them to an unlisted RBM3 isoform.
Which phosphorylation sites are annotated for RBM3?
PTM · UniProt lists phosphoserine at position 147 and phosphotyrosine at position 155. These are UniProt coordinates; check the numbering convention before comparing them with an antibody or paper. Their presence does not demonstrate a resolvable shift on a Western blot.
Does this guide establish induction of RBM3?
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 RBM3 Western blot?
Transfer · RBM3 is predicted to be 17.2 kDa, so check that your transfer conditions retain a protein of this size. The supplied features do not specify a membrane or transfer protocol. Confirm recovery near the observed 22 kDa band when optimizing transfer.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03104-2 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 RBM3 be quantified across cell fractions?
Quantitation · UniProt places RBM3 in the nucleus and cytoplasm and notes localization in dendritic mRNA granules. Compare like fractions and use an appropriate loading reference for each fraction. A change in one fraction alone need not represent a change in total RBM3.
Why does RBM3 appear near 22 kDa instead of 17.2 kDa?
Interpretation · The supplied Western blots show an apparent band near 22 kDa, versus a predicted mass of 17.2 kDa. RBM3 has annotated methylation and phosphorylation, but those features alone do not establish the cause of the difference. Use the observed band position when evaluating RBM3 signals.
UniProt annotates methylarginine at positions 47, 105, 121, and 131. Position 105 has alternate methylation annotations, including dimethylation reported in A2780 ovarian carcinoma cells. These are UniProt coordinates; antibody or paper numbering may differ. The annotations do not establish a visible band shift.
Start with the observed band near 22 kDa and assess other bands independently. The record lists one isoform, methylation, and phosphorylation, but does not establish that any modification produces a separate visible band. RBM3 also associates with RPL4 and 60S ribosomal subunits; that association alone does not identify an extra band.
UniProt annotates methylarginine at positions 47, 105, 121, and 131. Position 105 has alternate methylation annotations, including dimethylation reported in A2780 ovarian carcinoma cells. These are UniProt coordinates; antibody or paper numbering may differ. The annotations do not establish a visible band shift.
Start with the observed band near 22 kDa and assess other bands independently. The record lists one isoform, methylation, and phosphorylation, but does not establish that any modification produces a separate visible band. RBM3 also associates with RPL4 and 60S ribosomal subunits; that association alone does not identify an extra band.
Boster reagents
RBM3 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-RBM3 Antibody Picoband®
Cat # A03104-2
Real WB data
Anti-RBM3 Rabbit Monoclonal Antibody
Cat # M03104
Both the supplier anti-RBM3 antibodies have Western blot images from human cell lysates. A03104-2 shows a band near 22 kDa, above the expected 17 kDa; M03104 shows a band near 17 kDa. The supplied images do not show mouse samples.
Which to pick: For human samples, M03104 has an imaged band at the expected 17 kDa; A03104-2 shows a band near 22 kDa. M03104 also lists mouse reactivity, but its supplied Western blot image tests human lysates only.
Source: BosterBio RBM3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P98179.
- Human Protein Atlas. RBM3 tissue expression.
- PMC4570027 — target-verified WB comparison
- PMC5747149 — target-verified WB comparison
- PMC13079030 — target-verified WB comparison