This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
RPS15 / Small ribosomal subunit protein uS19 · Western blot design guide
Design a Western Blot for RPS15
Source-linked RPS15 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RPS15 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
RPS15 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 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Breast (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 | Acetylated + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked RPS15 Western Blot Protocol Options
The A06538 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | SK-OVCAR3 (40ug), HEK293T (40ug), A549 (40ug), AML-12 (40ug), PC12 (40ug) (catalog A06538) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A06538; 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 RPS15 Western Blot Band Size?
RPS15 is predicted at 17 kDa; its N-terminal acetylation is documented, but a migration effect and an empirical band size are not established.
| Single band near 17 kDa | consistent with the predicted size of RPS15 |
| Band near 17 kDa in whole-cell lysate | consistent with its cytoplasmic location |
| Faint band near 17 kDa | possible RPS15 signal; confirm with an identity control |
| No band near 17 kDa | RPS15 is not detected under these conditions |
💡Expected RPS15 appearanceRPS15 has a predicted mass of 17 kDa, but no empirical band size is supplied; a band near 17 kDa is plausible and should be confirmed with an antibody specificity control.
| Predicted molecular mass | sets a 17 kDa reference for the unmodified protein |
| 145-residue sequence | underlies the predicted molecular mass |
| N-acetylalanine at residue 2 | is documented, but no visible migration effect is established |
| Small ribosomal subunit association | does not establish a different denaturing gel band size |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | RPS15 is not detected despite its cytoplasmic location | check lysate loading and antibody performance with a positive-control lysate |
| Band higher than expected | the identity of the higher band is unestablished | compare with a molecular-weight marker and confirm specificity by RPS15 depletion |
| Band lower than expected | the identity of the lower band is unestablished | check sample integrity and confirm specificity by RPS15 depletion |
| Multiple bands | a single isoform is listed, so the additional bands have no established RPS15 explanation | use RPS15 depletion to identify the specific band |
| Weak or no signal | insufficient detectable RPS15 or ineffective detection | check loading and detection with a positive-control lysate |
| Fragments below expected size | sample degradation is possible; no cleavage feature is listed | prepare fresh lysate with protease inhibitors and compare with an RPS15 identity control |
Experiment evidence
Western blot images showing RPS15 band
A06538 · RPS15
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RPS15 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RPS15 in Western blot, you can use breast tissue, which shows high HPA expression.
Positive control: Breast (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: Cytoplasmic RPS15 is suitable for whole-cell lysate, but the adipose tissue negative should be validated.
HPA tissue expression evidence for RPS15
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 |
|---|---|---|---|---|
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | GLUC cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Colon | endocrine 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 → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | Low | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
Section 3
Advanced RPS15 Western Blot Tips
Deeper troubleshooting and optimisation questions for RPS15, answered from its protein features.
Where should the RPS15 band appear?
Band shift · RPS15 has a predicted mass of 17 kDa. No observed Western blot band is supplied, so use 17 kDa as a starting reference rather than an exact apparent mass.
Could RPS15 isoforms produce multiple bands?
Isoforms · Only one isoform is supplied, with no alternative sequence. The supplied features therefore do not support assigning multiple bands to RPS15 isoforms.
Can acetylation explain a shifted RPS15 band?
PTM · UniProt lists N-acetylalanine at position 2. If another source uses different residue numbering, compare its convention before assigning the site. This feature alone does not establish a visible band shift or explain a difference from 17 kDa.
Does this guide establish induction of RPS15?
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 RPS15 Western blot?
Transfer · RPS15 is a small, 17-kDa protein. Use a setup suitable for retaining small proteins and check transfer near 17 kDa experimentally. The supplied features do not specify a membrane or transfer time.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A06538 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 RPS15 be quantified across samples?
Quantitation · RPS15 is cytoplasmic and a component of the small ribosomal subunit. Compare matched sample fractions, load comparable total protein, and measure signals within the assay's linear range. The supplied features do not establish that RPS15 abundance is constant across conditions.
How should unexpected higher RPS15 bands be interpreted?
Interpretation · The supplied features list N-acetylalanine at UniProt position 2 and include a Ubl conjugation keyword, but give no conjugation site or expected band size. Do not assign a higher band to a specific modification from these features alone; verify band identity experimentally.
Boster reagents
RPS15 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-40S ribosomal protein S15 RPS15 Antibody
Cat # A06538
A06538 is a polyclonal anti-RPS15 antibody listed as reactive with human, mouse, and rat. Its WB image shows 40 µg whole-cell lysates from SK-OVCAR3, HEK293T, A549, AML-12, and PC12 at 1:500. No further validation is supplied.
Which to pick: A06538 is the only listed antibody and has a WB image. Choose it as a starting point for the named sample contexts; confirm performance with your own samples and conditions.
Source: BosterBio RPS15 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P62841.
- Human Protein Atlas. RPS15 tissue expression.
- PMC5088344 — target-verified WB comparison
- PMC6917584 — target-verified WB comparison
- PMC10235050 — target-verified WB comparison