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- Table of Contents
MRPL47 / Large ribosomal subunit protein uL29m · Western blot design guide
Design a Western Blot for MRPL47
Source-linked MRPL47 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MRPL47 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
MRPL47 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 | ~29.5 kDa | |
| Observed band | 72 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Ovary (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Acetylated | |
| Caveat | Peptide-blocking control | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked MRPL47 Western Blot Protocol Options
The A14516 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | the lysates from 293 cells (catalog A14516) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A14516; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected MRPL47 Western Blot Band Size?
MRPL47 is predicted at 29.5 kDa, while antibody QC reports 72 kDa; the cause of this difference is not established.
| Band at 72 kDa | empirical MRPL47 antibody band; its identity and migration cause require confirmation |
| Band near 29.5 kDa | consistent with the predicted protein mass, but identity requires confirmation |
| Several bands at different positions | isoforms 1, 2, and 3 are annotated, but distinct migration is unproven |
| Band below the full-length position | possible mitochondrial transit-peptide processing; cleavage extent is unspecified |
💡Expected MRPL47 appearanceMRPL47 has a predicted mass of 29.5 kDa, while antibody QC reports a 72 kDa band; the difference is unexplained, so confirm band identity with peptide blocking or another antibody.
| Predicted mass | 29.5 kDa by sequence; the reported antibody band is 72 kDa |
| Isoforms 1, 2, and 3 | could differ in size, but their masses and migration are unspecified |
| N6-acetyllysine at residue 144 | is annotated without evidence of a visible size shift |
| Mitochondrial transit peptide | could yield a smaller processed form, but no cleavage site or mature mass is supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | MRPL47 is mitochondrial and may be weak in total lysate | check mitochondrial enrichment and antibody performance |
| Band higher than expected | the reported 72 kDa band exceeds the 29.5 kDa prediction for an unknown reason | compare peptide-blocked and unblocked lanes and confirm with another antibody |
| Band lower than expected | transit-peptide processing or an isoform is possible, but neither migration is established | verify identity with an independent antibody and mitochondrial enrichment |
| Multiple bands | three isoforms are annotated, but their band positions are unknown | use peptide blocking and an independent antibody to identify specific bands |
| Weak or no signal | the mitochondrial target may be underrepresented in the sample | enrich mitochondria and verify sample loading |
| Fragments below expected size | protein degradation or processing may produce smaller species | compare fresh, protected samples and confirm bands with an independent antibody |
Experiment evidence
Western blot images showing MRPL47 band
A14516 · MRPL47
A14516 · MRPL47
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MRPL47 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MRPL47 in Western blot, you can use appendix tissue, which HPA rates High.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Ovary (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Ovary is a plausible negative control because HPA reports MRPL47 as not detected there.
HPA tissue expression evidence for MRPL47
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 → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Hippocampus | neuronal cells | Low | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced MRPL47 Western Blot Tips
Deeper troubleshooting and optimisation questions for MRPL47, answered from its protein features.
How should MRPL47 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 MRPL47 isoforms produce different bands?
Isoforms · UniProt lists three isoforms. Relative to isoform 1, isoform 2 lacks residues 34–53 and isoform 3 lacks residues 1–110. These sequence differences could affect migration, but the features do not predict their observed band positions.
Can acetylation explain the apparent band shift?
PTM · UniProt lists N6-acetyllysine at residue 144 in its sequence numbering. That modification alone does not demonstrate a visible shift or explain a band at 72 kDa. Keep the UniProt coordinate explicit when comparing it with antibody or publication numbering.
Does this guide establish induction of MRPL47?
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 MRPL47 Western blot?
Transfer · Check transfer and detection across both the predicted 29.5 kDa region and the reported 72 kDa region. The supplied features do not specify a membrane, buffer, or transfer method that would establish which band is MRPL47.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A14516 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 MRPL47 bands be quantified?
Quantitation · Quantify a consistently identified band, recording whether it is near the predicted 29.5 kDa mass or the reported 72 kDa position. Because UniProt lists three isoforms, do not combine distinct bands as one MRPL47 measurement without establishing their identities.
Does the 72 kDa band match MRPL47's predicted mass?
Interpretation · MRPL47 is predicted at 29.5 kDa, well below the reported 72 kDa apparent band. Its listed features do not establish the cause of that difference. Check band identity before assigning the 72 kDa signal to MRPL47.
Compare them with the predicted mass and the sequence differences among the three isoforms. MRPL47 is listed as a mitochondrial large ribosomal subunit component, but these features alone cannot identify an unexpected band or attribute it to acetylation.
Compare them with the predicted mass and the sequence differences among the three isoforms. MRPL47 is listed as a mitochondrial large ribosomal subunit component, but these features alone cannot identify an unexpected band or attribute it to acetylation.
Boster reagents
MRPL47 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-MRPL47 Antibody
Cat # A14516
The catalog reports one anti-MRPL47 antibody, A14516, with reported human, mouse, and rat reactivity. Its Western blot image is described as using 293 cell lysate; the supplied evidence does not establish Western blot performance across all listed species.
Which to pick: A14516 is the only listed option and has a Western blot image from 293 cell lysate. For other sample types, the supplied record provides no sample-specific Western blot evidence.
Source: BosterBio MRPL47 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9HD33.
- Human Protein Atlas. MRPL47 tissue expression.
- PMC12809090 — target-verified WB comparison
- PMC11853951 — target-verified WB comparison