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- Table of Contents
RPL6 / Large ribosomal subunit protein eL6 · Western blot design guide
Design a Western Blot for RPL6
Source-linked RPL6 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RPL6 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
RPL6 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 | ~32.7 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (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 | 1 isoform(s) |
Section 1
Source-Linked RPL6 Western Blot Protocol Options
The A07614 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A07614) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A07614) |
| Primary antibody | A07614 · 1:1000 (catalog A07614) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A07614) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A07614) |
Section 2
What Is the Expected RPL6 Western Blot Band Size?
RPL6 has a predicted mass of 32.7 kDa; its listed modifications could affect migration, but no shifted band is demonstrated.
| Single band near 32.7 kDa | Consistent with predicted RPL6 size; confirm identity with an RPL6 depletion control |
| Band near 32.7 kDa in a cytosolic fraction | Consistent with the reported cytosolic location of RPL6 |
| Band near 32.7 kDa in a polysome fraction | Consistent with detection of RPL6 on cytosolic polysomes |
| Band near 32.7 kDa in a rough ER fraction | Consistent with the reported rough ER association of RPL6 |
| Close doublet near 32.7 kDa | Could reflect modified RPL6 states; the listed modifications do not establish a visible doublet |
💡Expected RPL6 appearanceUniProt predicts RPL6 at 32.7 kDa; no empirical band size is supplied, and its listed modifications do not establish a migration shift, so confirm a candidate band with an RPL6 depletion control.
| UniProt predicted mass | Sets a reference of 32.7 kDa for RPL6 |
| N6-succinyllysine at Lys94 | May affect migration, but no visible size change is established |
| Phosphoserine at Ser127 | May affect migration, but no visible size change is established |
| N6-succinyllysine at Lys207 | May affect migration, but no visible size change is established |
| N6-acetyllysine at Lys239 | May affect migration, but no visible size change is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Insufficient detectable RPL6 or ineffective antibody detection | Check loading and transfer, then compare with a cytosolic or polysome fraction |
| Band higher than expected | A modified RPL6 state or an unrelated antibody target; neither is established | Check band identity with RPL6 depletion and compare treated and untreated samples where relevant |
| Band lower than expected | Possible protein breakdown or an unrelated antibody target | Prepare fresh lysate with protease inhibitors and check band identity by RPL6 depletion |
| Multiple bands | Possible modified RPL6 states or antibody cross-reactivity | Check which bands decrease after RPL6 depletion |
| Weak or no signal | Low loaded RPL6 amount or ineffective detection | Check loading, transfer and antibody conditions; compare a polysome-enriched fraction |
| Fragments below expected size | Possible degradation of RPL6 during preparation | Keep samples cold, use protease inhibitors and compare fresh lysate |
Experiment evidence
Western blot images showing RPL6 band
A07614 · RPL6
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RPL6 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RPL6 in Western blot, you can use appendix tissue, which HPA lists as a positive sample.
Positive control: Appendix (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: HPA lists adipose tissue as not detected, but verify that it lacks a band in your lysate.
HPA tissue expression evidence for RPL6
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 | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal 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 → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | follicle cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RPL6 Western Blot Tips
Deeper troubleshooting and optimisation questions for RPL6, answered from its protein features.
Where should the main RPL6 band appear?
Band shift · The predicted mass is 32.7 kDa. No empirical apparent band is supplied, so use this as a reference rather than an exact migration position.
Should multiple RPL6 isoform bands be expected?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore gives no isoform-based explanation for multiple bands; assess additional bands without assigning them to an unlisted isoform.
Can RPL6 modifications explain a shifted band?
PTM · UniProt lists N6-succinyllysine at 94 and 207, phosphoserine at 127, and N6-acetyllysine at 239. These are UniProt sequence coordinates; antibody or paper numbering may differ. Their presence alone does not establish a visible shift or explain a mass difference.
Does this guide establish induction of RPL6?
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 RPL6 Western blot?
Transfer · Plan transfer around the predicted 32.7 kDa protein and verify that RPL6 is recovered on the membrane. The supplied features do not specify a transfer method or a special transfer requirement.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A07614 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 RPL6 be quantified across samples?
Quantitation · Keep sample fractions consistent: RPL6 is a large ribosomal subunit component detected in cytosolic polysomes and ribosomes associated with rough endoplasmic reticulum. A change in the sampled fraction could affect the measured signal.
How should unexpected higher RPL6 bands be interpreted?
Interpretation · The record includes a Ubl conjugation keyword, but supplies no conjugation site or expected band size. Treat that as a hypothesis to test, not an identification of a higher band. The listed modified residues also do not establish a visible shift.
Boster reagents
RPL6 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-60S ribosomal protein L6 RPL6 Antibody
Cat # A07614
A07614 is the sole listed anti-RPL6 antibody. It is reported reactive with human, mouse, and rat and has a Western blot image of extracts from various cell lines. The caption does not identify those cell lines or their species.
Which to pick: A07614 is the only listed option. Its WB image uses 25 µg of cell-line extract per lane and a 1:1000 primary dilution. Check its reported human, mouse, and rat reactivity against your intended sample.
Source: BosterBio RPL6 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q02878.
- Human Protein Atlas. RPL6 tissue expression.
- PMC9899609 — target-verified WB comparison
- PMC12520468 — target-verified WB comparison
- PMC6825013 — target-verified WB comparison