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- Table of Contents
POLRMT / DNA-directed RNA polymerase, mitochondrial · Western blot design guide
Design a Western Blot for POLRMT
Source-linked POLRMT Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-POLRMT 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
POLRMT 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 | ~138.6 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked POLRMT Western Blot Protocol Options
The A06341 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 A06341) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A06341; 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 POLRMT Western Blot Band Size?
POLRMT has a predicted 138.6 kDa monomer; homodimer formation and a transit peptide annotation suggest possibilities, but no altered migration or empirical band size is established.
| Band near 138.6 kDa | Consistent with the predicted POLRMT monomer; confirm identity with antibody controls |
| Higher band near 277 kDa | Could reflect a homodimer if it survives sample preparation |
| Monomer and higher band together | Could reflect monomeric POLRMT and retained homodimer |
| Weak band in whole-cell lysate | Mitochondrial localization may make POLRMT easier to detect in a mitochondrial fraction |
💡Expected POLRMT appearanceUniProt predicts a 138.6 kDa monomer; no empirical band size is supplied, so confirm any band near that size with ordinary band-identity controls.
| UniProt predicted mass | Places the full-length monomer near 138.6 kDa |
| 1230-residue sequence | Underlies the predicted full-length mass; no separate mature mass is supplied |
| Homodimer formation | Could produce a band near twice the monomer size if the dimer survives sample preparation |
| Mitochondrial transit peptide annotation | No cleavage site or resulting size change is supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | POLRMT is mitochondrial and may be scarce in the sampled lysate | Check a mitochondrial-enriched fraction and a positive control |
| Band higher than expected | A POLRMT homodimer may persist during sample preparation | Compare denaturing sample conditions and confirm identity with an independent antibody |
| Band lower than expected | The supplied features do not establish a cleavage product or its mass | Check sample integrity and confirm the band with an independent antibody |
| Multiple bands | Monomer and a retained homodimer are possible; other band identities are unestablished | Compare denaturing conditions and verify bands with independent antibody controls |
| Weak or no signal | Mitochondrial localization may limit POLRMT abundance in whole-cell lysate | Enrich mitochondria and verify recovery with a mitochondrial marker |
Experiment evidence
Western blot images showing POLRMT band
A06341 · POLRMT
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for POLRMT Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for POLRMT in Western blot, you can use adrenal gland tissue, which HPA rates as high expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA lists no undetected tissue, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for POLRMT
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 | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | adipocytes | High | Protein (IHC) | HPA → |
| Caudate | glial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cervix | glandular cells | Medium | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced POLRMT Western Blot Tips
Deeper troubleshooting and optimisation questions for POLRMT, answered from its protein features.
Where should the POLRMT band appear relative to predicted mass?
Band shift · The predicted mass is 138.6 kDa. An empirical Western blot band position is unavailable, so use 138.6 kDa as a reference, not a confirmed apparent mass. The transit peptide keyword alone does not establish a processing site or explain a mass difference.
Could POLRMT isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands. Check band identity before assigning additional bands to POLRMT.
Do listed modifications predict a POLRMT band shift?
PTM · No modified residues or glycosylation sites are listed. The supplied features do not identify a modification that predicts a shift, but they cannot rule out unlisted modifications. Do not assign a shifted band to a specific modification from this record.
Does this guide establish induction of POLRMT?
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 POLRMT Western blot?
Transfer · At a predicted 138.6 kDa, POLRMT is relatively large. A wet transfer is a practical starting point; verify transfer near 139 kDa with a molecular weight marker before interpreting a weak or absent band.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A06341 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 POLRMT be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
How should POLRMT abundance be compared across samples?
Interpretation · POLRMT is mitochondrial. When comparing bands, account for differences in mitochondrial content between samples and measure signal within the assay's linear range. The supplied features do not establish an induction condition.
POLRMT is described as a homodimer, but that feature alone does not establish that a dimer survives Western blot preparation. Treat a high band as unassigned until its POLRMT identity is independently supported.
POLRMT is described as a homodimer, but that feature alone does not establish that a dimer survives Western blot preparation. Treat a high band as unassigned until its POLRMT identity is independently supported.
Boster reagents
POLRMT 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-POLRMT Antibody
Cat # A06341
the supplier A06341 is an anti-POLRMT antibody listed for human reactivity. Its Western blot image is captioned as analysis of extracts from various cell lines; specific lines, conditions, and publication evidence are not supplied.
Which to pick: A06341 is the only listed option for POLRMT. It has a Western blot image and listed human reactivity; check the full product details for suitability to your sample and protocol.
Source: BosterBio POLRMT gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O00411.
- Human Protein Atlas. POLRMT tissue expression.
- PMC11680805 — target-verified WB comparison
- PMC9569504 — target-verified WB comparison
- PMC13202344 — target-verified WB comparison