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- Table of Contents
TUFM / Elongation factor Tu, mitochondrial · Western blot design guide
Design a Western Blot for TUFM
Source-linked TUFM Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TUFM 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
TUFM 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 | ~49.9 kDa | |
| Gel | 12–15% (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 | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked TUFM Western Blot Protocol Options
The A30672 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A30672; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | 1:10000 (catalog A30672) |
| 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 TUFM Western Blot Band Size?
TUFM has a predicted 49.9 kDa precursor; transit peptide processing and listed modifications may affect migration, but no mature or empirical band size is supplied.
| Band near 49.9 kDa | Consistent with the predicted TUFM precursor mass; confirm identity with controls |
| Band below 49.9 kDa | Could reflect mitochondrial transit peptide processing; mature mass is unspecified |
| Several bands near the expected size | Could reflect modified TUFM states; distinct migration is unproven |
| Faint lysate band with a stronger mitochondrial fraction band | Consistent with TUFM localization to the mitochondrial matrix |
💡Expected TUFM appearanceUniProt predicts a 49.9 kDa TUFM precursor; the transit peptide and listed modifications do not establish a mature band size, so confirm any candidate band with identity controls.
| UniProt predicted mass | Places the 455-residue precursor near 49.9 kDa |
| Mitochondrial transit peptide | Processing may yield a smaller mature protein; no cleavage site or mature mass is supplied |
| Lysine acetylation at residues 82, 91 and 259 | Modified states are listed, but a visible size shift is not established |
| Lysine succinylation at residues 91, 237 and 289 | Modified states are listed, but a visible size shift is not established |
| Phosphorylation at residues 281 and 315 | Modified states are listed, but distinct migration is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TUFM resides in the mitochondrial matrix | Check mitochondrial enrichment and lysate recovery |
| Band higher than expected | The supplied features do not establish a higher-mass species | Check molecular-weight markers and confirm band identity with an independent antibody |
| Band lower than expected | Mitochondrial transit peptide processing is possible | Compare mitochondrial and whole-cell samples and confirm band identity |
| Multiple bands | Modified states are annotated, but their separation is unproven | Compare an independent antibody and TUFM-depleted control |
| Weak or no signal | Mitochondrial matrix protein may be poorly recovered | Verify mitochondrial recovery and sample loading |
| Fragments below expected size | Fragment identity is not established by the supplied features | Check sample integrity and use an independent antibody |
Experiment evidence
Western blot images showing TUFM band
A30672 · TUFM
A05606-2 · TUFM
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TUFM Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TUFM in Western blot, you can use adrenal gland tissue.
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 such as Ponceau alongside.
⚠️Feasibility: No clean negative tissue is reported; use siRNA knockdown or a KO line.
HPA tissue expression evidence for TUFM
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 | endocrine cells | High | Protein (IHC) | HPA → |
| Breast | myoepithelial cells | High | Protein (IHC) | HPA → |
| Bronchus | basal cells | High | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Fallopian tube | ciliated cells (cell body) | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Caudate | glial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Medium | Protein (IHC) | HPA → |
Section 3
Advanced TUFM Western Blot Tips
Deeper troubleshooting and optimisation questions for TUFM, answered from its protein features.
How should TUFM 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 a second band be a TUFM isoform?
Isoforms · Only one isoform is listed, with no alternative sequence. The supplied features do not support assigning a second band to an annotated TUFM isoform.
Which lysine modifications should I consider when interpreting TUFM bands?
PTM · UniProt lists acetyllysine at positions 82, 259, 364, and 421; succinyllysine at 237 and 289; and alternate acetyllysine or succinyllysine at 91. These are UniProt coordinates, which may differ from paper or antibody numbering. None establishes a visible band shift.
UniProt lists phosphothreonine at position 281 and phosphoserine at 315. Check these UniProt coordinates against the numbering used for any phospho-specific antibody. Their presence does not establish a shifted band or a condition that induces phosphorylation.
UniProt lists phosphothreonine at position 281 and phosphoserine at 315. Check these UniProt coordinates against the numbering used for any phospho-specific antibody. Their presence does not establish a shifted band or a condition that induces phosphorylation.
Does this guide establish induction of TUFM?
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 TUFM?
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 A30672 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 TUFM 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 I interpret a band away from 49.9 kDa?
Interpretation · 49.9 kDa is the predicted mass; no observed band position is supplied. TUFM has a transit peptide annotation and several modified residues, but these features alone do not establish a visible shift or explain a particular apparent mass.
TUFM is annotated as a mitochondrial matrix protein with a transit peptide, but no cleavage position is supplied. Do not assign a specific mature mass or processing-related band from these features.
TUFM is annotated in the mitochondrial matrix. For a total-TUFM comparison, apply the same sample preparation and loading approach across samples and quantify the same band consistently. The supplied features do not identify an induced condition or establish that modified forms resolve as separate bands.
Compare it with the 49.9 kDa predicted mass and the annotated mitochondrial location. A transit peptide and modified residues are listed, but no cleavage coordinate or empirical band position is supplied. These features cannot establish the identity of an unexpected band.
TUFM is annotated as a mitochondrial matrix protein with a transit peptide, but no cleavage position is supplied. Do not assign a specific mature mass or processing-related band from these features.
TUFM is annotated in the mitochondrial matrix. For a total-TUFM comparison, apply the same sample preparation and loading approach across samples and quantify the same band consistently. The supplied features do not identify an induced condition or establish that modified forms resolve as separate bands.
Compare it with the 49.9 kDa predicted mass and the annotated mitochondrial location. A transit peptide and modified residues are listed, but no cleavage coordinate or empirical band position is supplied. These features cannot establish the identity of an unexpected band.
Boster reagents
TUFM 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-TUFM Antibody
Cat # A30672
Real WB data
Anti-TUFM Antibody Picoband®
Cat # A05606-2
The catalog reports two anti-TUFM antibodies, A30672 and A05606-2, with human, mouse, and rat reactivity and WB images. A05606-2’s caption reports an approximately 50 kDa band (46 kDa expected); A30672’s supplied cue gives dilutions but no named specimens.
Which to pick: For the documented human cell lines and rat or mouse tissues, consider A05606-2: its WB caption identifies the samples and conditions. A30672 also has a WB image, but its supplied cue describes only various cell lysates.
Source: BosterBio TUFM gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P49411.
- Human Protein Atlas. TUFM tissue expression.
- PMC13330442 — target-verified WB comparison
- PMC9348918 — target-verified WB comparison
- PMC13577116 — target-verified WB comparison