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- Table of Contents
RBM39 / RNA-binding protein 39 · Western blot design guide
Design a Western Blot for RBM39
Source-linked RBM39 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RBM39 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
RBM39 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 | ~59.4 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Liver (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 | 3 isoform(s) |
Section 1
Source-Linked RBM39 Western Blot Protocol Options
The A07137 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A07137) |
| 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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A07137) |
| Primary antibody | A07137 · 1:1000 (catalog A07137) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A07137) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A07137) |
Section 2
What Is the Expected RBM39 Western Blot Band Size?
RBM39 is predicted at 59.4 kDa; isoforms and phosphorylation could affect migration, but no empirical band size or visible effect is established.
| Band near 59.4 kDa | consistent with the predicted RBM39 mass; identity needs confirmation |
| Slightly shifted band or doublet | could reflect annotated phosphorylation; a visible shift is unproven |
| Several bands at different positions | could include RBM39 isoforms 1, 2, and 3; distinct migration is unproven |
| Weak band in whole-cell lysate | RBM39 is concentrated in nuclear speckles |
💡Expected RBM39 appearanceRBM39 has a predicted mass of 59.4 kDa, but no empirical band size is supplied; confirm a candidate band with an RBM39 depletion or independent antibody control.
| Predicted RBM39 mass | 59.4 kDa is the sequence-based reference, not a measured band position |
| Isoforms 1, 2, and 3 | alternative splicing could change size, but their relative masses and migration are unspecified |
| Phosphotyrosine at residue 95 | could affect migration; no visible shift is established |
| Phosphoserine at residues 97 and 100 | could affect migration; no visible shift is established |
| N-acetylalanine at residue 2 | adds a small modification without an established visible shift |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | RBM39 is concentrated in nuclear speckles, and the sample may contain too little detectable RBM39 | check a nuclear-enriched fraction and a positive cell lysate |
| Band higher than expected | annotated phosphorylation could affect migration, but its effect is unverified | compare phosphatase-treated and untreated samples and confirm band identity by RBM39 depletion |
| Band lower than expected | an RBM39 isoform may differ in size, but isoform masses are unavailable | check antibody epitope coverage and confirm the band by RBM39 depletion |
| Multiple bands | isoforms or differing phosphorylation states are possible; distinct bands are not established | compare phosphatase treatment and identify RBM39-dependent bands with depletion |
| Weak or no signal | a whole-cell sample may dilute nuclear-speckle RBM39 | test a nuclear-enriched fraction and verify antibody performance with a positive control |
Experiment evidence
Western blot images showing RBM39 band
A07137 · RBM39
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RBM39 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RBM39 in Western blot, you can use appendix tissue lysate, which HPA rates as high.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Liver (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA lists liver as not detected, but confirm its background signal in your assay.
HPA tissue expression evidence for RBM39
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 | glandular cells | High | Protein (IHC) | HPA → |
| Caudate | neuronal cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Thyroid gland | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced RBM39 Western Blot Tips
Deeper troubleshooting and optimisation questions for RBM39, answered from its protein features.
How should RBM39 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 RBM39 isoforms produce different bands?
Isoforms · UniProt lists three isoforms. Relative to the canonical sequence, isoform 3 lacks residues 100–121 and isoform 2 lacks residues 392–397. These deletions could affect band position, but the features do not establish which isoforms are present in your sample.
Isoform 3 lacks canonical residues 100–121, including the annotated phosphoserines at 100, 117, and 121. Check whether a phosphosite antibody targets that region before interpreting an isoform 3 signal. The positions given here use UniProt canonical numbering; antibody or paper numbering may differ.
Isoform 3 lacks canonical residues 100–121, including the annotated phosphoserines at 100, 117, and 121. Check whether a phosphosite antibody targets that region before interpreting an isoform 3 signal. The positions given here use UniProt canonical numbering; antibody or paper numbering may differ.
Which RBM39 modifications matter when interpreting a blot?
PTM · In UniProt canonical numbering, RBM39 has N-acetylalanine at position 2; phosphotyrosine at 95; phosphoserine at 97, 100, 117, 121, 136, 334, 337, and 341; and phosphothreonine at 146. Compare phosphosite labels using the same numbering convention. These annotations alone do not predict a visible band shift.
Does this guide establish induction of RBM39?
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 RBM39?
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 A07137 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 RBM39 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.
Should RBM39 migrate at its predicted 59.4 kDa?
Interpretation · Use 59.4 kDa as the predicted reference. No observed band size is supplied, and the listed modifications do not establish a visible shift or explain any difference from the predicted mass.
RBM39 is concentrated in nuclear speckles. Keep sample preparation and nuclear enrichment consistent across lanes, then compare signals within the same validated band or isoform. If multiple bands appear, measure them separately until their identities are established.
Check whether bands are compatible with the isoform 3 deletion of canonical residues 100–121 or the isoform 2 deletion of 392–397. The supplied features do not establish an observed band size or prove that either deletion or a modification causes a particular band. Verify band identity before assigning it to RBM39.
RBM39 is concentrated in nuclear speckles. Keep sample preparation and nuclear enrichment consistent across lanes, then compare signals within the same validated band or isoform. If multiple bands appear, measure them separately until their identities are established.
Check whether bands are compatible with the isoform 3 deletion of canonical residues 100–121 or the isoform 2 deletion of 392–397. The supplied features do not establish an observed band size or prove that either deletion or a modification causes a particular band. Verify band identity before assigning it to RBM39.
Boster reagents
RBM39 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-RNA-binding protein 39 RBM39 Antibody
Cat # A07137
A07137 is listed for RBM39 with reported Human, Mouse, and Rat reactivity and a Western blot image. Its caption describes cell-line extracts tested at 1:1000 with 25 µg per lane; the cell lines are unnamed.
Which to pick: A07137 is the only listed antibody. Consider it for samples within its reported reactivity, using the pictured Western blot conditions as a starting point. The caption does not identify which cell lines or species were tested.
Source: BosterBio RBM39 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q14498.
- Human Protein Atlas. RBM39 tissue expression.
- PMC11995665 — target-verified WB comparison
- PMC11609384 — target-verified WB comparison
- PMC9095732 — target-verified WB comparison