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- Table of Contents
RBPMS / RNA-binding protein with multiple splicing · Western blot design guide
Design a Western Blot for RBPMS
Source-linked RBPMS Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RBPMS WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
RBPMS 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 | ~21.8 kDa | |
| Observed band | ~24 and 28 kDa | |
| Gel | 12% (catalog A07130-2) | |
| Positive control | Colon (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 | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 5 isoform(s) |
Section 1
Source-Linked RBPMS Western Blot Protocol Options
The A07130-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Caco-2, human A549, human RT4, human HepG2, rat heart, mouse heart (catalog A07130-2) |
| Gel % | 12% (catalog A07130-2) |
| Load | 30 ug; reducing conditions (catalog A07130-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A07130-2) |
| Membrane | nitrocellulose membrane (catalog A07130-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A07130-2) |
| Primary antibody | A07130-2 · 0.5 μg/mL (catalog A07130-2) |
| Primary incubation | overnight at 4°C (catalog A07130-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A07130-2) |
| Secondary incubation | 1.5 hour at RT (catalog A07130-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A07130-2) |
| Detection | ECL (catalog A07130-2) |
Section 2
What Is the Expected RBPMS Western Blot Band Size?
RBPMS has a predicted mass of 21.8 kDa and empirical bands at approximately 24 and 28 kDa; the cause of the difference is not established.
| Bands at approximately 24 and 28 kDa | Empirical RBPMS bands in reducing whole-cell and tissue lysates; their difference is unexplained |
| Band near 21.8 kDa | Near the predicted mass of the supplied sequence |
| Several bands at different positions | Could reflect isoforms A, B, C, D or E; their migration is not established |
| Band near 44 kDa after incomplete denaturation | Could reflect retained RBPMS homodimer |
💡Expected RBPMS appearanceUniProt predicts 21.8 kDa, while antibody QC shows bands at approximately 24 and 28 kDa; their identities and the reason for the difference need band-identity controls.
| Predicted 21.8 kDa sequence mass | Provides a reference for the approximately 24 and 28 kDa empirical bands, without explaining their migration |
| Isoforms A, B, C, D and E | May differ in size, but their masses and separation on a blot are not supplied |
| Homodimer formation | Could yield a band near twice the monomer mass if the complex survives sample preparation |
| Phosphothreonine at residue 12 | May affect migration; no visible size shift is established |
| Phosphothreonine at residue 113 | May affect migration; no visible size shift is established |
| N-acetylmethionine at residue 1 | Is a documented modification; no visible size shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | The observed 24 and 28 kDa bands exceed the 21.8 kDa sequence mass; the cause is unknown | Compare with a validated RBPMS antibody and RBPMS depletion control |
| Band lower than expected | An unverified band may be an isoform or a fragment | Check antibody epitope coverage and test RBPMS depletion |
| Multiple bands | Isoforms A through E exist, but their band positions are unknown | Use RBPMS depletion and isoform-specific controls to assign bands |
| Weak or no signal | RBPMS distribution between nucleus and cytoplasm may affect the sampled fraction | Check nuclear and cytoplasmic fractions and verify extraction and loading |
| Fragments below expected size | Protein breakdown during sample preparation is possible; no fragment mass is supplied | Prepare fresh lysate with protease inhibitors and check RBPMS dependence |
Experiment evidence
Western blot images showing RBPMS band
A07130-2 · RBPMS
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RBPMS Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RBPMS in Western blot, you can use colon tissue, which HPA scores High.
Positive control: Colon (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: A tissue control pair is feasible, but lysates should retain both nuclear and cytoplasmic proteins.
HPA tissue expression evidence for RBPMS
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 |
|---|---|---|---|---|
| Colon | endothelial cells | High | Protein (IHC) | HPA → |
| Kidney | cells in glomeruli | High | Protein (IHC) | HPA → |
| Lung | alveolar cells type II | High | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | High | Protein (IHC) | HPA → |
| Urinary bladder | urothelial cells | 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 → |
| 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 → |
| Cerebral cortex | endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RBPMS Western Blot Tips
Deeper troubleshooting and optimisation questions for RBPMS, answered from its protein features.
Why might RBPMS bands appear near 24 and 28 kDa?
Band shift · The canonical sequence predicts 21.8 kDa, while the supplied Western blots show bands near 24 and 28 kDa. RBPMS has alternative isoforms and listed modifications, but those features alone do not establish the cause of either apparent mass. Check which isoforms the antibody recognizes before assigning the bands.
Could RBPMS isoforms produce different bands?
Isoforms · UniProt lists isoforms A–E. Isoform D replaces residues 133–196, E lacks 133–176, and B and C have different sequences at 177–196. These changes could affect band positions or antibody recognition. Check the antibody epitope against each isoform before interpreting multiple bands.
Which RBPMS modifications should I consider when interpreting bands?
PTM · UniProt lists N-acetylmethionine at position 1 and phosphothreonine at positions 12 and 113. These are UniProt coordinates, which may differ from paper or antibody numbering. Their presence does not by itself demonstrate a visible band shift; use a phosphatase comparison if testing whether phosphorylation contributes to a band difference.
Does this guide establish induction of RBPMS?
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 RBPMS Western blot?
Transfer · The canonical RBPMS sequence predicts a 21.8 kDa protein. Choose transfer conditions that retain and detect proteins around that size, and verify transfer with a suitable molecular-weight marker and membrane stain. The supplied UniProt features do not establish one preferred transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A07130-2 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 RBPMS 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 I compare RBPMS amounts across samples?
Interpretation · RBPMS is listed in both nucleus and cytoplasm, including stress granules and P-bodies. Keep fractionation and loading consistent across samples, and quantify the same validated band or set of bands each time. If the antibody recognizes multiple isoforms, state whether the measurement represents one band or their combined signal.
First check whether the antibody epitope is present in isoforms A–E, particularly across the altered region spanning residues 133–196. Compare bands with the supplied 24 and 28 kDa observations, but do not assign identity from apparent mass alone. The listed modifications and homodimerization also do not, by themselves, identify an unexpected Western-blot band.
UniProt places RBPMS in the nucleus and cytoplasm and also lists cytoplasmic stress granules and P-bodies. Use the same extraction approach across samples when comparing signal. If testing subcellular distribution, analyze fractions separately and verify their purity before interpreting differences in RBPMS bands.
First check whether the antibody epitope is present in isoforms A–E, particularly across the altered region spanning residues 133–196. Compare bands with the supplied 24 and 28 kDa observations, but do not assign identity from apparent mass alone. The listed modifications and homodimerization also do not, by themselves, identify an unexpected Western-blot band.
UniProt places RBPMS in the nucleus and cytoplasm and also lists cytoplasmic stress granules and P-bodies. Use the same extraction approach across samples when comparing signal. If testing subcellular distribution, analyze fractions separately and verify their purity before interpreting differences in RBPMS bands.
Boster reagents
RBPMS 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-RBPMS Antibody Picoband®
Cat # A07130-2
A07130-2 is a rabbit polyclonal anti-RBPMS antibody listed for human, mouse, and rat. Its Western blot image uses four human cell lysates plus rat and mouse heart lysates. Reported bands at approximately 24 and 28 kDa differ from the expected 22 kDa.
Which to pick: A07130-2 is the only listed option and has a Western blot image. Choose it when the reported human cell or rodent heart samples fit your experiment; the image does not establish performance in every tissue or condition.
Source: BosterBio RBPMS gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q93062.
- Human Protein Atlas. RBPMS tissue expression.
- PMC8745614 — target-verified WB comparison
- PMC9116735 — target-verified WB comparison
- PMC3959221 — target-verified WB comparison