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- Table of Contents
RPS12 / Small ribosomal subunit protein eS12 · Western blot design guide
Design a Western Blot for RPS12
Source-linked RPS12 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RPS12 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
RPS12 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 | ~14.5 kDa | |
| Observed band | ~15 kDa | |
| Gel | 10% (catalog A01040-3) | |
| Positive control | Bronchus (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 | Acetylated | |
| Caveat | — | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked RPS12 Western Blot Protocol Options
The A01040-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human MCF-7, human HepG2 (catalog A01040-3) |
| Gel % | 10% (catalog A01040-3) |
| Load | 30 ug; reducing conditions (catalog A01040-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01040-3) |
| Membrane | nitrocellulose membrane (catalog A01040-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01040-3) |
| Primary antibody | A01040-3 · 1:1000 (catalog A01040-3) |
| Primary incubation | overnight at 4°C (catalog A01040-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01040-3) |
| Secondary incubation | 1.5 hour at RT (catalog A01040-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01040-3) |
| Detection | ECL (catalog A01040-3) |
Section 2
What Is the Expected RPS12 Western Blot Band Size?
RPS12 is predicted at 14.5 kDa and observed at ~15 kDa; the cause of the small difference is not established.
| ~15 kDa band in whole-cell lysate | matches the empirically observed RPS12 band |
| band near 14.5 kDa | consistent with the predicted RPS12 mass |
| ~15 kDa band in a cytoplasmic fraction | consistent with RPS12 cytoplasmic localization |
| ~15 kDa band in a nucleolar fraction | consistent with RPS12 nucleolar localization |
💡Expected RPS12 appearanceRPS12 has a predicted mass of 14.5 kDa and an observed whole-cell lysate band at ~15 kDa; confirm band identity with ordinary antibody specificity controls.
| UniProt predicted mass | predicts a band near 14.5 kDa; the observed band is ~15 kDa |
| N-acetylalanine at residue 2 | no visible migration effect is established |
| N6-succinyllysine at residue 129 | no visible migration effect is established |
| Single annotated isoform | provides no basis for assigning additional isoform band sizes |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | RPS12 may be lost during recovery of cytoplasmic or nucleolar proteins | check extraction, loading, and a positive whole-cell lysate |
| Band higher than expected | the listed features do not establish a higher-mass RPS12 species | verify band identity with an independent antibody or RPS12 depletion |
| Band lower than expected | no signal peptide or propeptide cleavage is annotated | check band identity with an independent antibody or RPS12 depletion |
| Multiple bands | the single annotated isoform does not explain multiple bands | compare antibody specificity controls and RPS12 depletion |
| Weak or no signal | incomplete recovery or transfer of this small protein | check lysate loading, transfer, and a positive whole-cell lysate |
Experiment evidence
Western blot images showing RPS12 band
A01040-3 · RPS12
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RPS12 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RPS12 in Western blot, you can use bronchus tissue, which HPA rates as high expression.
Positive control: Bronchus (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: Whole-cell lysate is suitable for this intracellular protein; verify the HPA not-detected tissue as a negative control.
HPA tissue expression evidence for RPS12
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 |
|---|---|---|---|---|
| Bronchus | basal cells | High | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | glandular cells | High | 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 → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RPS12 Western Blot Tips
Deeper troubleshooting and optimisation questions for RPS12, answered from its protein features.
Where should the RPS12 band appear?
Band shift · RPS12 has a predicted mass of 14.5 kDa, and the supplied observed band is approximately 15 kDa. Use that region when identifying the band; apparent mobility need not match calculated mass exactly.
Could an RPS12 isoform explain another band?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for an additional band.
Do the listed modifications explain a band shift?
PTM · UniProt lists N-acetylalanine at position 2 and N6-succinyllysine at position 129. These are UniProt sequence coordinates; antibody or paper numbering may differ. Their presence alone does not establish a visible shift or explain a difference between observed and predicted mass.
The supplied record lists no glycosylation sites. It provides no basis for assigning a higher band to glycosylated RPS12.
The supplied record lists no glycosylation sites. It provides no basis for assigning a higher band to glycosylated RPS12.
Does this guide establish induction of RPS12?
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 RPS12?
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 A01040-3 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.
What matters when quantifying RPS12 bands?
Quantitation · Quantify a consistently identified band near the supplied approximately 15 kDa position. The record does not establish that other bands represent RPS12 or that either listed modification produces a separately quantifiable band.
How should unexpected RPS12 bands be interpreted?
Interpretation · Compare them with the approximately 15 kDa observed band. The listed modifications and single isoform do not, by themselves, identify another band as RPS12; confirm band identity before interpreting it.
UniProt locates RPS12 in the cytoplasm and the nucleus, including the nucleolus. Consider both locations when interpreting fractionated samples; RPS12 is also listed as a subunit of the 40S ribosomal complex.
UniProt locates RPS12 in the cytoplasm and the nucleus, including the nucleolus. Consider both locations when interpreting fractionated samples; RPS12 is also listed as a subunit of the 40S ribosomal complex.
Boster reagents
RPS12 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-RPS12 Antibody Picoband®
Cat # A01040-3
The catalog reports one anti-RPS12 antibody, A01040-3, with a Western blot image showing an approximately 15 kDa band in the specified human, rat, and mouse cell lysates. This is product image evidence under the reported conditions, not broader validation.
Which to pick: A01040-3 is the only listed option. Its WB image uses HeLa, MCF-7, HepG2, K562, PC-12, and NIH/3T3 whole cell lysates, with 30 µg per lane and a 1:1000 primary dilution. Match your sample context to those reported.
Source: BosterBio RPS12 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P25398.
- Human Protein Atlas. RPS12 tissue expression.
- PMC10287158 — target-verified WB comparison
- PMC12632061 — target-verified WB comparison
- PMC3854201 — target-verified WB comparison