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- Table of Contents
RPL23 / Large ribosomal subunit protein uL14 · Western blot design guide
Design a Western Blot for RPL23
Real validated RPL23 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RPL23 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
RPL23 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~14.9 kDa | |
| Observed band | ~17 kDa | |
| Gel | 5–20% (catalog A06155-2) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Smooth muscle (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
Real Curated RPL23 Western Blot Protocols
The A06155-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human PC-3, human THP-1, rat stomach (catalog A06155-2) |
| Gel % | 5–20% (catalog A06155-2) |
| Load | 30 ug; reducing conditions (catalog A06155-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A06155-2) |
| Membrane | nitrocellulose membrane (catalog A06155-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A06155-2) |
| Primary antibody | A06155-2 · 0.5 μg/mL (catalog A06155-2) |
| Primary incubation | overnight at 4°C (catalog A06155-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A06155-2) |
| Secondary incubation | 1.5 hour at RT (catalog A06155-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A06155-2) |
| Detection | ECL (catalog A06155-2) |
Section 2
What Is the Expected RPL23 Western Blot Band Size?
RPL23 is predicted at 14.9 kDa and observed near 17 kDa; the cause of the difference is not established.
| Band near 17 kDa | Matches the empirical RPL23 band; confirm identity with controls |
| Band near 14.9 kDa | Near the UniProt sequence-predicted mass; identity needs confirmation |
| Additional close band near the main band | Could reflect a different phosphorylation state involving Ser17 or Tyr38; separation is unproven |
| No band in whole-cell lysate | Unexpected for cytoplasmic RPL23; check detection and sample handling |
💡Expected RPL23 appearanceUniProt predicts 14.9 kDa, while antibody QC shows a band near 17 kDa in reducing whole-cell lysates; the cause of the difference is unestablished, so confirm identity with appropriate controls.
| UniProt predicted mass | 14.9 kDa from the protein sequence |
| Phosphoserine at Ser17 | May affect mobility, but no visible shift is established |
| Phosphotyrosine at Tyr38 | May affect mobility, but no visible shift is established |
| Phosphoprotein annotation | Modification-state effects on apparent size remain unquantified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | RPL23 is cytoplasmic, so absence suggests a sample or detection problem | Check lysate loading and a positive control |
| Band higher than expected | The observed 17 kDa band exceeds the 14.9 kDa prediction for an unestablished reason | Compare with the QC band and confirm identity by RPL23 depletion |
| Band lower than expected | A lower band has no established feature-based explanation | Check sample integrity and confirm identity by RPL23 depletion |
| Multiple bands | Phosphorylation states or unrelated antibody binding are possible; resolved forms are unproven | Compare with RPL23 depletion and consider a phosphatase-treated control |
| Weak or no signal | Insufficient detectable RPL23 or poor transfer may limit signal | Check loading, transfer, and a positive lysate control |
Experiment evidence
Western blot images showing RPL23 band
A06155-2 · RPL23
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RPL23 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RPL23 in Western blot, you can use appendix tissue, which shows high HPA expression.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Smooth muscle (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Smooth muscle offers a tissue negative, while several other tissues show high expression.
HPA tissue expression evidence for RPL23
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 → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Low | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Low | Protein (IHC) | HPA → |
| Soft tissue | chondrocytes | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
Section 3
Advanced RPL23 Western Blot Tips
Deeper troubleshooting and optimisation questions for RPL23, answered from its protein features.
How should RPL23 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.
Should different RPL23 bands be assigned to isoforms?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It provides no basis for assigning additional bands to RPL23 isoforms; confirm their identity before doing so.
Which RPL23 phosphorylation sites should a phospho-specific assay target?
PTM · UniProt lists phosphoserine at position 17 and phosphotyrosine at position 38. These are UniProt coordinates; check the numbering convention of any antibody or paper before matching sites. A total-RPL23 assay alone cannot establish phosphorylation at either site.
Phosphorylation is listed at Ser17 and Tyr38, but site presence alone does not demonstrate a resolvable band shift. Compare the band with a suitable phosphorylation-sensitive assay before attributing it to either modification.
Phosphorylation is listed at Ser17 and Tyr38, but site presence alone does not demonstrate a resolvable band shift. Compare the band with a suitable phosphorylation-sensitive assay before attributing it to either modification.
Does this guide establish induction of RPL23?
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 RPL23 Western blot?
Transfer · RPL23 is a small protein with a predicted mass of 14.9 kDa and an observed band near 17 kDa. Choose transfer conditions validated for retention of proteins in that size range, and check transfer before interpreting a weak or absent band.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A06155-2 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 RPL23 bands be quantified across samples?
Quantitation · Quantify the identified band near 17 kDa using comparable sample loading and exposures within the assay’s linear range. RPL23 is a cytoplasmic component of the large ribosomal subunit, so interpret changes in its signal in that biological context.
Why might RPL23 appear near 17 kDa instead of 14.9 kDa?
Interpretation · 14.9 kDa is the predicted mass, while the supplied Western blot observation is approximately 17 kDa. The listed phosphorylation sites do not establish the cause of this difference or demonstrate a visible shift. Use the observed position as a guide and confirm band identity experimentally.
The record lists one isoform, phosphorylation at Ser17 and Tyr38, and no signal peptide, propeptide, or glycosylation sites. None of these features alone identifies an extra band. Confirm unexpected bands independently before assigning them to RPL23 or a specific modification.
The record lists one isoform, phosphorylation at Ser17 and Tyr38, and no signal peptide, propeptide, or glycosylation sites. None of these features alone identifies an extra band. Confirm unexpected bands independently before assigning them to RPL23 or a specific modification.
Boster reagents
RPL23 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-RPL23 Antibody Picoband®
Cat # A06155-2
The catalog reports one anti-RPL23 antibody for Western blot, A06155-2, with stated human, mouse, and rat reactivity. Its WB image shows a band near the expected 17 kDa in human PC-3 and THP-1 cells and rat stomach tissue; mouse WB testing is not shown.
Which to pick: A06155-2 is the only listed option. Its WB image supports the reported human cell and rat tissue conditions. For mouse samples, reactivity is listed, but the supplied WB image does not show a mouse sample.
Source: BosterBio RPL23 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P62829.
- Human Protein Atlas. RPL23 tissue expression.
- PMC5443795 — target-verified WB comparison
- PMC7160537 — target-verified WB comparison
- PMC8677661 — target-verified WB comparison