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- Table of Contents
RPL11 / Large ribosomal subunit protein uL5 · Western blot design guide
Design a Western Blot for RPL11
Real validated RPL11 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RPL11 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
RPL11 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 | ~20.3 kDa | |
| Observed band | ~18 kDa | |
| Gel | 5–20% (catalog A02901-1) | |
| Positive control | Adrenal gland (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 C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated RPL11 Western Blot Protocols
The A02901-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human A549, human Jurkat, human Raji (catalog A02901-1) |
| Gel % | 5–20% (catalog A02901-1) |
| Load | 30 ug; reducing conditions (catalog A02901-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02901-1) |
| Membrane | nitrocellulose membrane (catalog A02901-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02901-1) |
| Primary antibody | A02901-1 · 0.5 μg/mL (catalog A02901-1) |
| Primary incubation | overnight at 4°C (catalog A02901-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02901-1) |
| Secondary incubation | 1.5 hour at RT (catalog A02901-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02901-1) |
| Detection | ECL (catalog A02901-1) |
Section 2
What Is the Expected RPL11 Western Blot Band Size?
RPL11 is predicted at 20.3 kDa but observed at ~18 kDa; the cause of the difference is not established.
| Band at ~18 kDa | Matches the empirical RPL11 band in reducing whole-cell blots; confirm identity with controls. |
| Band near 20.3 kDa | Near the sequence-predicted mass; identity requires confirmation. |
| Multiple bands | Isoforms 1 and 2 are annotated, but distinct band positions are not established. |
| Bands in nuclear and cytoplasmic fractions | Consistent with RPL11 localization in both compartments. |
💡Expected RPL11 appearanceRPL11 has a predicted mass of 20.3 kDa, while reducing whole-cell blots show a band at ~18 kDa; the cause of the difference is unestablished, so confirm identity with ordinary band-identity controls.
| Predicted sequence mass of 20.3 kDa | Provides a size reference; the empirical band is ~18 kDa, with no established cause for the difference. |
| Alternative splicing | Produces annotated isoforms 1 and 2; their relative sizes and migration are not supplied. |
| Isoform 1 | Has no supplied isoform-specific mass or established band position. |
| Isoform 2 | Has no supplied isoform-specific mass or established band position. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | RPL11 recovery or detection may be inadequate. | Check lysate loading, transfer and a positive-control lysate. |
| Band higher than expected | Its identity or migration is unconfirmed; annotated modifications do not establish a visible shift. | Compare with a positive control and verify antibody specificity. |
| Band lower than expected | The ~18 kDa empirical band runs below the 20.3 kDa prediction for an unestablished reason. | Compare with the validated blot and confirm band identity. |
| Multiple bands | Isoforms 1 and 2 or annotated modifications are possible, but separate band positions are unestablished. | Confirm each band with an independent antibody or RPL11 depletion. |
| Weak or no signal | Insufficient RPL11 recovery or detection may limit signal. | Check loading and transfer, then test a positive-control lysate. |
| Fragments below expected size | Sample degradation is possible; no cleavage product is specified. | Prepare fresh lysate with protease inhibitors and compare intact-protein signal. |
Experiment evidence
Western blot images showing RPL11 band
A02901-1 · RPL11
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RPL11 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RPL11 in Western blot, you can use adrenal gland lysate, which HPA scores as medium expression.
Positive control: Adrenal gland (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: Adipose tissue is reported as not detected, but confirm its negative signal in your blot.
HPA tissue expression evidence for RPL11
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial 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 → |
| Kidney | cells in glomeruli | Not detected | Protein (IHC) | HPA → |
| Ovary | follicle cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RPL11 Western Blot Tips
Deeper troubleshooting and optimisation questions for RPL11, answered from its protein features.
How should RPL11 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.
Can isoform 2 explain a separate RPL11 band?
Isoforms · UniProt lists isoforms 1 and 2; isoform 2 lacks residue 3 in UniProt numbering. That small sequence difference alone does not establish a resolvable second band. Confirm band identity before assigning a band to either isoform.
Which RPL11 modifications matter when interpreting bands?
PTM · UniProt lists N-acetylalanine at position 2, phosphothreonine at positions 44 and 47, alternate N6-acetyllysine at position 52, and N6-acetyllysine at position 85. These are UniProt coordinates, which may differ from paper or antibody numbering. Their presence does not establish a visible band shift.
Check whether the assay targets UniProt Thr44 or Thr47, the two listed phosphothreonine sites. State the numbering convention when comparing antibody or paper labels. A total RPL11 band cannot identify which site is phosphorylated.
Check whether the assay targets UniProt Thr44 or Thr47, the two listed phosphothreonine sites. State the numbering convention when comparing antibody or paper labels. A total RPL11 band cannot identify which site is phosphorylated.
Does this guide establish induction of RPL11?
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 RPL11?
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 A02901-1 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 RPL11 be quantified across samples?
Quantitation · Quantify the same verified RPL11 band across comparable samples. Record whether measurements use whole-cell, nuclear, or cytoplasmic material: UniProt lists RPL11 in the nucleolus and cytoplasm. Do not combine additional bands into the measurement without verifying their identity.
Why does RPL11 appear near 18 kDa instead of 20.3 kDa?
Interpretation · The supplied apparent band is ~18 kDa, while the predicted mass is 20.3 kDa. The listed features do not establish the cause of this difference. Use the observed position when identifying the band, and verify its identity if other bands appear.
Check the expected ~18 kDa band first. Isoform 2 lacks only residue 3, and the listed modifications do not by themselves establish extra visible bands. RPL11 is part of the large ribosomal subunit and 5S RNP complex, but these features alone cannot identify an unexpected band.
Check the expected ~18 kDa band first. Isoform 2 lacks only residue 3, and the listed modifications do not by themselves establish extra visible bands. RPL11 is part of the large ribosomal subunit and 5S RNP complex, but these features alone cannot identify an unexpected band.
Boster reagents
RPL11 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-RPL11 Antibody Picoband®
Cat # A02901-1
The catalog reports A02901-1 for RPL11 Western blotting with reported human, mouse, and rat reactivity. Its WB image shows an approximately 18 kDa band in human cell lines and mouse cell and liver lysates; the supplied image does not test rat samples.
Which to pick: A02901-1 is the only listed option. Its WB image includes A549, Jurkat, Raji, HeLa, NIH/3T3, and mouse liver lysates. For rat samples, reactivity is listed, but no rat WB example is supplied.
Source: BosterBio RPL11 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P62913.
- Human Protein Atlas. RPL11 tissue expression.
- PMC3406587 — target-verified WB comparison
- PMC7038965 — target-verified WB comparison