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RPS26 / Small ribosomal subunit protein eS26 · Western blot design guide
Design a Western Blot for RPS26
Source-linked RPS26 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RPS26 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
RPS26 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 | ~13 kDa | |
| Observed band | ~18 kDa | |
| Gel | 8% (catalog A06454) | |
| Positive control | Gallbladder (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 | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked RPS26 Western Blot Protocol Options
The A06454 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human 293T, human HEL, human Caco-2, human PC-3, rat lung, rat ovary, mouse lung, mouse ovary (catalog A06454) |
| Gel % | 8% (catalog A06454) |
| Load | 30 ug; reducing conditions (catalog A06454) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A06454) |
| Membrane | nitrocellulose membrane (catalog A06454) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A06454) |
| Primary antibody | A06454 · 1:1000 (catalog A06454) |
| Primary incubation | overnight at 4°C (catalog A06454) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A06454) |
| Secondary incubation | 1.5 hour at RT (catalog A06454) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A06454) |
| Detection | ECL (catalog A06454) |
Section 2
What Is the Expected RPS26 Western Blot Band Size?
RPS26 is predicted at 13 kDa and observed near 18 kDa in the supplied blots; the cause of the difference is not established.
| Band near 18 kDa | Matches the empirical RPS26 band in the supplied whole-cell blots; the cause of its migration is unknown |
| Band near 13 kDa | Matches the predicted RPS26 mass; confirm its identity |
| Little signal in a soluble fraction | Ribosome-associated RPS26 may have been lost during fractionation |
| Closely spaced bands | Ser54 phosphorylation is possible, but distinct migration is unproven |
💡Expected RPS26 appearanceRPS26 is predicted at 13 kDa, while the supplied whole-cell blots show a band near 18 kDa; confirm band identity because the cause of the difference is not established.
| Predicted RPS26 mass | 13 kDa from UniProt, compared with the approximately 18 kDa empirical band |
| 115-residue sequence | Defines the predicted full-length protein; no cleavage is listed |
| Ser54 phosphorylation | May affect migration, but no visible size shift is demonstrated |
| 40S ribosomal subunit association | Can affect recovery during fractionation; no change in SDS-PAGE monomer size is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Ribosome-associated RPS26 may have been lost during fractionation | Check the retained pellet and whole-cell lysate |
| Band higher than expected | The supplied 18 kDa band exceeds the 13 kDa prediction for an unknown reason | Compare with a validated RPS26 control and confirm antibody specificity |
| Band lower than expected | The supplied features do not establish a smaller RPS26 species | Check sample integrity and confirm band identity |
| Multiple bands | Ser54 phosphorylation is documented, but distinct bands are not established | Confirm bands with an independent RPS26 antibody or depletion control |
| Weak or no signal | Ribosome-associated RPS26 may be depleted from the prepared fraction | Compare whole-cell lysate with the discarded fraction |
Experiment evidence
Western blot images showing RPS26 band
A06454 · RPS26
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RPS26 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RPS26 in Western blot, you can use gallbladder lysate, which has high HPA expression.
Positive control: Gallbladder (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: Gallbladder is a strong positive candidate, but verify that adipose tissue lysate lacks detectable RPS26 signal.
HPA tissue expression evidence for RPS26
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 |
|---|---|---|---|---|
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
| Liver | cholangiocytes | High | Protein (IHC) | HPA → |
| Pancreas | exocrine glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular 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 → |
| Epididymis | glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Kidney | cells in glomeruli | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RPS26 Western Blot Tips
Deeper troubleshooting and optimisation questions for RPS26, answered from its protein features.
How should RPS26 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 different RPS26 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. Check band specificity before labeling additional bands as RPS26.
Which RPS26 phosphorylation site should I consider?
PTM · UniProt lists phosphoserine at position 54, using UniProt sequence numbering. Antibody or paper numbering may differ, so align sequences before comparing site labels. The listed modification does not by itself predict a resolvable Western-blot band shift.
Does this guide establish induction of RPS26?
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 RPS26 Western blot?
Transfer · RPS26 is a small, 115-residue protein with a predicted mass of 13 kDa. Choose transfer conditions and a membrane that retain small proteins, then check the membrane and gel after transfer to assess retention and completeness.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A06454 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 RPS26 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.
Why might RPS26 appear near 18 kDa instead of 13 kDa?
Interpretation · The reported apparent band is about 18 kDa, while the predicted mass is 13 kDa. UniProt lists phosphoserine at position 54, but that feature alone does not establish a visible shift or explain the difference. Identify the band with an independent specificity check before assigning its cause.
RPS26 is a component of the 40S subunit and is detected on cytosolic polysomes. Compare equivalent sample fractions and equal protein loads; differences in ribosome content or fraction preparation can affect the RPS26 signal.
The record lists one isoform, no signal or propeptide, no glycosylation sites, and phosphoserine at UniProt position 54. These features do not establish the identity of an extra band. Verify antibody specificity and compare the band across consistent cytosolic or ribosome-containing preparations.
RPS26 is a component of the 40S subunit and is detected on cytosolic polysomes. Compare equivalent sample fractions and equal protein loads; differences in ribosome content or fraction preparation can affect the RPS26 signal.
The record lists one isoform, no signal or propeptide, no glycosylation sites, and phosphoserine at UniProt position 54. These features do not establish the identity of an extra band. Verify antibody specificity and compare the band across consistent cytosolic or ribosome-containing preparations.
Boster reagents
RPS26 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-RPS26 Antibody
Cat # A06454
The catalog reports one anti-RPS26 antibody, A06454, with a Western blot image from human cell, rat tissue, and mouse tissue lysates. The reported band is approximately 18 kDa, versus an expected 13 kDa; the caption does not resolve this discrepancy.
Which to pick: A06454 is the only listed option. Its stated reactivity covers human, mouse, and rat, and its WB image includes human 293T, HEL, Caco-2, and PC-3 cells plus rat and mouse lung and ovary tissues. Check the 18 kDa band against your expected result.
Source: BosterBio RPS26 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P62854.
- Human Protein Atlas. RPS26 tissue expression.
- PMC12084008 — target-verified WB comparison
- PMC6242890 — target-verified WB comparison
- PMC10201367 — target-verified WB comparison