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- Table of Contents
RPL18 / Large ribosomal subunit protein eL18 · Western blot design guide
Design a Western Blot for RPL18
Source-linked RPL18 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RPL18 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
RPL18 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 | ~21.6 kDa | |
| Observed band | ~23 kDa | |
| Gel | 5–20% (catalog A07057-3) | |
| Positive control | Testis (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 + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked RPL18 Western Blot Protocol Options
The A07057-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human 293T, human HepG2, human Daudi, rat testis, rat brain, mouse testis, mouse brain (catalog A07057-3) |
| Gel % | 5–20% (catalog A07057-3) |
| Load | 30 ug; reducing conditions (catalog A07057-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A07057-3) |
| Membrane | nitrocellulose membrane (catalog A07057-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A07057-3) |
| Primary antibody | A07057-3 · 0.5 μg/mL (catalog A07057-3) |
| Primary incubation | overnight at 4°C (catalog A07057-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A07057-3) |
| Secondary incubation | 1.5 hour at RT (catalog A07057-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A07057-3) |
| Detection | ECL (catalog A07057-3) |
Section 2
What Is the Expected RPL18 Western Blot Band Size?
RPL18 is predicted at 21.6 kDa and observed at ~23 kDa; the cause of the difference is not established.
| Band at ~23 kDa | Matches the empirical RPL18 band in reducing whole-cell lysates |
| Band near 21.6 kDa | Near the UniProt predicted mass; confirm its identity |
| Two bands near the expected region | Isoforms 1 and 2 are possible contributors, but their migration is unknown |
| Band at ~23 kDa across cell and tissue lysates | Consistent with the reported RPL18 blot across those samples |
💡Expected RPL18 appearanceUniProt predicts 21.6 kDa, while antibody QC reports an ~23 kDa band in reducing whole-cell lysates; the difference is unexplained, so confirm identity with appropriate controls.
| UniProt predicted mass | Sets a 21.6 kDa sequence-based reference |
| Isoform 1 | Has no supplied isoform-specific mass or migration |
| Isoform 2 | May differ in size from isoform 1, but its mass is not supplied |
| Alternative splicing | Could produce different band positions; resolvable bands are not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytosolic RPL18 was not detected despite its expected presence in whole-cell lysate | Check lysate loading and detection, then compare with a positive-control lysate |
| Band higher than expected | The reported ~23 kDa migration exceeds the 21.6 kDa prediction; its cause is unknown | Compare with the reported band and confirm identity using RPL18 depletion |
| Band lower than expected | An alternative isoform is possible, but no isoform migration is supplied | Confirm identity by RPL18 depletion and assess isoform expression |
| Multiple bands | Isoforms 1 and 2 are annotated, but distinct bands are unverified | Test which bands respond to RPL18 depletion |
| Weak or no signal | Insufficient detectable RPL18 in the prepared lysate or an assay problem | Check loading and transfer against the reported positive-control lysates |
Experiment evidence
Western blot images showing RPL18 band
A07057-3 · RPL18
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RPL18 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RPL18 in Western blot, you can use testis tissue, which HPA scores High.
Positive control: Testis (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 reports RPL18 as not detected in adipose tissue, but confirm that the negative sample lacks a detectable band.
HPA tissue expression evidence for RPL18
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 |
|---|---|---|---|---|
| Testis | elongated or late spermatids | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje 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 | Low | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Low | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Low | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
Section 3
Advanced RPL18 Western Blot Tips
Deeper troubleshooting and optimisation questions for RPL18, answered from its protein features.
How should RPL18 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 RPL18 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. In isoform 2, the canonical first 30 residues are replaced by a single methionine, so a smaller band is plausible. The sequence difference alone does not establish that both isoforms are expressed or resolved on a blot.
Isoform 2 replaces canonical residues 1–30 with one methionine. An antibody recognizing a sequence within those 30 residues may miss isoform 2. Check the antibody’s stated epitope before interpreting a missing or weaker isoform band.
Isoform 2 replaces canonical residues 1–30 with one methionine. An antibody recognizing a sequence within those 30 residues may miss isoform 2. Check the antibody’s stated epitope before interpreting a missing or weaker isoform band.
How could phosphorylation affect RPL18 band interpretation?
PTM · UniProt lists phosphoserine at position 130 and phosphothreonine at position 158, using canonical sequence coordinates. Compare matched samples with and without phosphatase treatment to test whether phosphorylation affects mobility. These sites do not by themselves predict a visible shift.
Does this guide establish induction of RPL18?
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 RPL18 Western blot?
Transfer · RPL18 is predicted at 21.6 kDa, with an observed band near 23 kDa. Use transfer conditions validated for retaining proteins in that size range, and check transfer around the 20–25-kDa markers. The supplied features do not specify a particular membrane or transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A07057-3 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 RPL18 be quantified across sample fractions?
Quantitation · RPL18 is a large ribosomal subunit component detected on cytosolic polysomes; the supplied location also includes rough endoplasmic reticulum. Compare like fractions and use a consistent loading measure, since fraction composition can affect the amount detected.
Why is the RPL18 band near 23 kDa instead of 21.6 kDa?
Interpretation · The observed ~23-kDa band is close to the predicted 21.6 kDa. The listed features do not establish why apparent and calculated masses differ. Phosphorylation at UniProt positions 130 and 158 alone does not demonstrate a visible shift.
First consider isoform 2, whose first 30 canonical residues are replaced by one methionine. Check whether the antibody can recognize the sequence retained in isoform 2, then compare the band with the expected ~23-kDa signal. The supplied features alone cannot identify an unexpected band.
First consider isoform 2, whose first 30 canonical residues are replaced by one methionine. Check whether the antibody can recognize the sequence retained in isoform 2, then compare the band with the expected ~23-kDa signal. The supplied features alone cannot identify an unexpected band.
Boster reagents
RPL18 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-RPL18 Antibody Picoband®
Cat # A07057-3
The catalog lists one anti-RPL18 antibody, A07057-3. Its WB image shows an approximately 23 kDa band (expected 22 kDa) in the stated human, rat, and mouse lysates. No publication or independent validation evidence is supplied.
Which to pick: A07057-3 is the only listed option. Its WB caption documents human HeLa, 293T, HepG2, and Daudi cell lysates, plus rat and mouse testis and brain lysates, using 30 µg sample and 0.5 µg/mL primary antibody.
Source: BosterBio RPL18 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q07020.
- Human Protein Atlas. RPL18 tissue expression.
- PMC9519341 — target-verified WB comparison
- PMC8245541 — target-verified WB comparison
- PMC8254245 — target-verified WB comparison