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- Table of Contents
RPL14 / Large ribosomal subunit protein eL14 · Western blot design guide
Design a Western Blot for RPL14
Source-linked RPL14 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RPL14 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
RPL14 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 | ~23.4 kDa | |
| Observed band | ~26 kDa | |
| Gel | 5–20% (catalog A07781-2) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
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 | 1 isoform(s) |
Section 1
Source-Linked RPL14 Western Blot Protocol Options
The A07781-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human 293T, human HepG2, human Hela, human MCF-7, rat liver, mouse liver, mouse NIH/3T3 (catalog A07781-2) |
| Gel % | 5–20% (catalog A07781-2) |
| Load | 30 ug; reducing conditions (catalog A07781-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A07781-2) |
| Membrane | nitrocellulose membrane (catalog A07781-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A07781-2) |
| Primary antibody | A07781-2 · 0.5 μg/mL (catalog A07781-2) |
| Primary incubation | overnight at 4°C (catalog A07781-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A07781-2) |
| Secondary incubation | 1.5 hour at RT (catalog A07781-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A07781-2) |
| Detection | ECL (catalog A07781-2) |
Section 2
What Is the Expected RPL14 Western Blot Band Size?
RPL14 is predicted at 23.4 kDa and observed near 26 kDa in reducing whole-cell lysates; the cause of the difference is not established.
| Band near 26 kDa | Empirical RPL14 band in reducing whole-cell lysates; confirm identity with antibody controls |
| Band near 23.4 kDa | Near the predicted RPL14 mass; identity needs confirmation |
| Close doublet near the main band | Modified RPL14 species are possible, but distinct migration is unproven |
| Additional higher band | A ubiquitin-like conjugated species is possible, but its migration is unestablished |
💡Expected RPL14 appearanceRPL14 has a predicted mass of 23.4 kDa and an empirical band near 26 kDa in reducing whole-cell lysates; the cause of the difference is unestablished, so confirm band identity with controls.
| Predicted RPL14 mass | 23.4 kDa from the UniProt sequence; the empirical band is near 26 kDa |
| Acetyllysine at residues 79 and 85 | May alter migration, but a visible shift is not established |
| Succinyllysine at residues 85 and 204 | May alter migration, but a visible shift is not established |
| Phosphoserine at residue 139 | May alter migration, but a visible shift is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytoplasmic RPL14 should be available in whole-cell lysate; loss during preparation or detection is possible | Check lysate loading and transfer against a known positive whole-cell lysate |
| Band higher than expected | The empirical RPL14 band is near 26 kDa despite the 23.4 kDa predicted mass; the cause is unestablished | Compare with the reported positive lysates and confirm antibody specificity |
| Band lower than expected | A band near 23.4 kDa could match the predicted mass, but its identity is unconfirmed | Compare with the 26 kDa reference band and verify specificity |
| Multiple bands | Modified RPL14 species are possible, but distinct bands are unproven | Use antibody specificity controls to identify the RPL14 band |
| Weak or no signal | Cytoplasmic RPL14 may be lost through insufficient lysate loading or transfer | Check protein loading, transfer, and a positive whole-cell lysate |
Experiment evidence
Western blot images showing RPL14 band
A07781-2 · RPL14
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RPL14 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RPL14 in Western blot, you can use appendix tissue lysate, which has high HPA expression.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: No tissue is reported as not detected, so use siRNA knockdown or a KO line for a negative control.
HPA tissue expression evidence for RPL14
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 → |
| Bone marrow | hematopoietic 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 → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Medium | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Medium | Protein (IHC) | HPA → |
Section 3
Advanced RPL14 Western Blot Tips
Deeper troubleshooting and optimisation questions for RPL14, answered from its protein features.
How should RPL14 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 RPL14 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore offers no isoform-based explanation for multiple bands. Investigate band identity before assigning additional bands to RPL14.
Which RPL14 modifications should I consider when interpreting bands?
PTM · UniProt lists acetyllysine at residue 79, alternate acetyllysine or succinyllysine at residue 85, phosphoserine at residue 139, and succinyllysine at residue 204. These are UniProt sequence coordinates; antibody or paper numbering may differ. Their presence does not establish a visible band shift.
UniProt marks N6-acetyllysine and N6-succinyllysine at residue 85 as alternate modifications. Interpret them as alternatives at that UniProt coordinate, not simultaneous modifications of one molecule.
UniProt marks N6-acetyllysine and N6-succinyllysine at residue 85 as alternate modifications. Interpret them as alternatives at that UniProt coordinate, not simultaneous modifications of one molecule.
Does this guide establish induction of RPL14?
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 RPL14?
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 A07781-2 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.
Which band should be quantified for RPL14 comparisons?
Quantitation · Quantify the same candidate RPL14 band across comparable samples, using the supplied approximately 26 kDa observation as a guide. The 23.4 kDa predicted mass alone does not establish which detected band is RPL14; verify band identity before including additional bands.
Why might RPL14 appear near 26 kDa instead of 23.4 kDa?
Interpretation · RPL14 has a predicted mass of 23.4 kDa, while the supplied Western blot observation is approximately 26 kDa. Its listed modifications do not establish the cause of that difference. Use the observed position to identify a candidate band, then verify its identity experimentally.
The supplied record lists no signal peptide or propeptide, so it provides no documented cleavage event that would explain a smaller band. Treat a smaller band as unassigned until its identity is checked.
UniProt places RPL14 in the cytoplasm and identifies it as a component of the large ribosomal subunit. Include material containing cytoplasmic ribosomes when assessing whether the approximately 26 kDa band is RPL14.
The supplied record lists no signal peptide or propeptide, so it provides no documented cleavage event that would explain a smaller band. Treat a smaller band as unassigned until its identity is checked.
UniProt places RPL14 in the cytoplasm and identifies it as a component of the large ribosomal subunit. Include material containing cytoplasmic ribosomes when assessing whether the approximately 26 kDa band is RPL14.
Boster reagents
RPL14 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-RPL14 Antibody Picoband®
Cat # A07781-2
The catalog reports one anti-RPL14 antibody, A07781-2, with stated human, mouse, and rat reactivity. Its WB image shows human cell, rat liver, and mouse liver/cell lysates, with a band near 26 kDa versus the expected 23 kDa. Evidence is limited to this product image.
Which to pick: A07781-2 is the only listed option and has a WB image. Its documented human cell, rat liver, and mouse liver/cell samples make those contexts practical starting points. Check the approximately 26 kDa band in your own sample.
Source: BosterBio RPL14 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P50914.
- Human Protein Atlas. RPL14 tissue expression.
- PMC7829600 — target-verified WB comparison
- PMC11921274 — target-verified WB comparison
- PMC8806664 — target-verified WB comparison